Antisense nucleic acids that enable exon skipping

Abstract

The present invention provides a pharmaceutical product for treating patients with various mutations by simultaneously skipping multiple exons (exon groups) of the target pre-mRNA. [Solution] An antisense oligomer, a pharmaceutically acceptable salt thereof, or a hydrate thereof is provided, which simultaneously skips two or more consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA in numerical order.

Description

[Technical Field] 【0001】 The present invention relates to an antisense oligomer capable of simultaneously skipping multiple exons of a target gene, and a pharmaceutical composition containing the oligomer. [Background technology] 【0002】 In recent years, exon skipping therapy, which treats diseases by skipping exons of genes containing disease-causing mutations to produce partially functional proteins, has attracted attention. Duchenne muscular dystrophy (DMD) is an example of a disease that can be treated with such exon skipping therapy. 【0003】 DMD is the most common hereditary progressive muscle disorder, affecting approximately 1 in 3,500 male births. While DMD patients exhibit almost the same motor function as healthy individuals in infancy, muscle weakness begins around the age of 4-5. Subsequently, muscle weakness in DMD patients progresses with age, leading to inability to walk by around age 12, and death from heart failure or respiratory failure in their 20s. Therefore, the development of effective treatments is urgently needed. 【0004】 It is known that the cause of DMD is a mutation in the dystrophin gene. The dystrophin gene is located on the X chromosome and is a huge gene consisting of 2.2 million base pairs of DNA. It is transcribed from DNA into pre-mRNA, and then spliced ​​to remove introns, resulting in mRNA with 79 exons joined together, making a total of 13,993 base pairs. This mRNA is then translated into 3,685 amino acids to produce the dystrophin protein. The dystrophin protein is involved in maintaining the membrane stability of muscle cells and is necessary to make muscle cells less susceptible to damage. Because DMD patients have a mutation in the dystrophin gene, functional dystrophin protein is hardly expressed in their muscle cells. As a result, in the bodies of DMD patients, the structure of muscle cells cannot be maintained during muscle contraction, and a large amount of calcium ions flow into the muscle cells. Consequently, muscle cell necrosis and fibrosis progress, and muscle cells gradually become less able to regenerate. 【0005】 Becker muscular dystrophy (BMD) is also caused by mutations in the dystrophin gene, but its symptoms, while presenting with muscle weakness, are generally milder than those of DMD, the progression of muscle weakness is slower, and it often develops in adulthood. The difference in clinical symptoms between DMD and BMD is thought to be due to whether the amino acid reading frame used when dystrophin mRNA is translated into dystrophin protein is disrupted or maintained by the mutation (Non-Patent Literature 1). In other words, in DMD, a mutation that shifts the amino acid reading frame results in almost no expression of functional dystrophin protein, while in BMD, although a portion of the exon is deleted due to the mutation, the amino acid reading frame is maintained, resulting in the production of an incomplete but functional dystrophin protein. 【0006】 Exon skipping is a promising treatment for DMD. This method involves modifying splicing to repair the amino acid reading frame of dystrophin mRNA, thereby inducing the expression of a partially functional dystrophin protein (Non-Patent Literature 2). The portion of the amino acid sequence translated from the exon targeted for exon skipping is lost. As a result, the dystrophin protein expressed by this treatment will be shorter than normal, but the amino acid reading frame is maintained, partially preserving its function in stabilizing muscle cells. Therefore, it is expected that exon skipping will cause DMD to present with symptoms similar to those of milder BMD. Exon skipping has undergone animal experiments in mice and dogs, and clinical trials are currently underway in human DMD patients. 【0007】 Exon skipping can be induced by the binding of antisense nucleic acids to either or both of the 5' or 3' splice sites, or to the interior of the exon. An exon is only incorporated into mRNA if both splice sites are recognized by the spliceosome complex. Therefore, exon skipping can be induced by targeting the area around the splice site with antisense nucleic acids. Furthermore, it is thought that the binding of serine and arginine-rich SR proteins to exon splicing enhancers (ESEs) is necessary for exons to be recognized by the splicing mechanism, and therefore, exon skipping can also be induced by targeting ESEs. 【0008】 Because mutations in the dystrophin gene vary among DMD patients, antisense nucleic acids are needed that are tailored to the location and type of gene mutation. There are several reports on antisense nucleic acids that induce exon skipping by targeting a single consecutive base sequence to a single exon of the dystrophin gene (Patent Documents 1-6 and Non-Patent Documents 1 and 2). Furthermore, it has been reported that mixing two types of antisense nucleic acids that target the same exon of the dystrophin gene (dual targeting) may enhance skipping activity compared to using each antisense nucleic acid alone (Patent Document 7). 【0009】 Furthermore, instead of skipping a single exon as described above, a method called multi-exon skipping, which skips multiple exons (or groups of exons), is attracting attention. This method makes it possible to treat a wide range of mutations in the dystrophin gene through exon skipping. For example, in the dystrophin gene, exons 45-55 are known as a hotspot for gene mutations, and it has been reported that by skipping these 11 exons, approximately 60% of DMD patients with deletion mutations can be treated (Non-Patent Literature 3). Also, while congenital deletion of exons 45-55 leads to BMD, it is known that most patients are asymptomatic or have mild symptoms (Non-Patent Literature 4). For these reasons, drugs that can induce exon 45-55 skipping are expected to be promising as DMD treatments. 【0010】 Methods for inducing multi-exon skipping include, for example, using antisense nucleic acids that target all exons in the region to be exon-skipped (Non-Patent Documents 5, 7, 8, and 10), and using antisense nucleic acids that target two different exons on the 3' and 5' sides of the region to be exon-skipped (Non-Patent Documents 6 and 9 and Patent Document 8). 【0011】 However, there has been no report yet that an antisense nucleic acid targeting a region including the vicinity of the donor or acceptor of an intron induces multi-exon skipping. 【Prior Art Documents】 【Patent Documents】 【0012】 【Patent Document 1】 International Publication Gazette No. 2004 / 048570 【Patent Document 2】 International Publication Gazette No. 2009 / 139630 【Patent Document 3】 International Publication Gazette No. 2010 / 048586 【Patent Document 4】 US Patent Publication Gazette No. 2010 / 0168212 【Patent Document 5】 International Publication Gazette No. 2011 / 057350 【Patent Document 6】 International Publication Gazette No. 2006 / 000057 【Patent Document 7】 International Publication Gazette No. 2007 / 135105 【Patent Document 8】 International Publication Gazette No. 2004 / 083446 【Non-Patent Documents】 【0013】 【Non-Patent Document 1】 Annemieke Aartsma-Rus et al., (2002) Neuromuscular Disorders 12: S71-S77 【Non-Patent Document 2】 Wilton S. D., e t al., Molecular Therapy 2007: 15: p. 1288-96 【Non-Patent Document 3】 Christophe Beroud et al., Human Mutation, 28(2), 2007, 196-202 【Non-Patent Document 4】 Yusuke Echigoya et al., Molecular Therapy‐Nucleic Acids, 4(2), 2015, e225 [Non-Patent Document 5] Yoshitsugu Aoki et al., PNAS, 109(34), 2012, 13763-13768 [Non-Patent Document 6] Laura van Vliet et al., BMC Medical Genetics, 9, 105, 2008 [Non-Patent Document 7] Joshua Lee et al., PLoS ONE, 13(5), e0197084, 2018 [Non-Patent Document 8] Joshua Lee et al., Methods in Molecular Biology, 1828, 141-150, 2018 [Non-Patent Document 9] Annemieke Aartsma-Rus et al, Am. J. Hum. Genet. 74(1), 83-92, 2004 [Non-Patent Document 10] Yusuke Echigoya et al., Molecular Therapy, 27(11), 1-13, 2019 [Overview of the Initiative] [Problems that the invention aims to solve] 【0014】 In the circumstances described above, there was a need for a drug that could treat patients with various mutations by simultaneously skipping multiple exons (or groups of exons) of the target pre-mRNA. [Means for solving the problem] 【0015】 The inventors, after conducting detailed studies of the technical details described in the above-mentioned literature and the structure of the dystrophin gene, discovered that an antisense nucleic acid targeting a specific region of human dystrophin pre-mRNA can simultaneously skip multiple exons from exons 45 to 55. Based on this finding, the inventors completed the present invention. 【0016】 In other words, the present invention is as follows: [1] An antisense oligomer, a pharmaceutically acceptable salt thereof, or a hydrate thereof, which simultaneously skips two or more consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA, The aforementioned antisense oligomer is The region Rn (where n is an odd number from 1 to 23) of the human dystrophin pre-mRNA, consisting of a sequence of NX bases from the 3' end to the 5' end of the NA exon and a sequence of NY bases from the 5' end to the 3' end of the NB intron, and The region Rn (where n is an even number from 2 to 24) of the aforementioned human dystrophin pre-mRNA consists of a sequence of NX bases from the 3' end to the 5' end of the NA intron and a sequence of NY bases from the 5' end to the 3' end of the NB exon. The sequence includes a nucleotide sequence that is complementary to the nucleotide sequence of at least one region selected from the group consisting of regions R1 to R24, which are shown as R1, R24, and R24, respectively. Here, When n=1, are NA=44, NB=44, NX=20, and NY=400? When n=2, are NA=44, NB=45, NX=600, and NY=50? When n=3, are NA=45, NB=45, NX=20, and NY=400? When n=4, are NA=45, NB=46, NX=400, and NY=50? When n=5, are NA=46, NB=46, NX=20, and NY=400? When n=6, are NA=46, NB=47, NX=400, and NY=50? When n=7, are NA=47, NB=47, NX=20, and NY=400? When n=8, are NA=47, NB=48, NX=400, and NY=50? When n=9, are NA=48, NB=48, NX=20, and NY=400? When n=10, are NA=48, NB=49, NX=400, and NY=50? When n=11, are NA=49, NB=49, NX=20, and NY=400? When n=12, are NA=49, NB=50, NX=400, and NY=50? When n=13, are NA=50, NB=50, NX=20, and NY=400? When n=14, are NA=50, NB=51, NX=400, and NY=50? When n=15, are NA=51, NB=51, NX=20, and NY=400? When n=16, are NA=51, NB=52, NX=400, and NY=50? When n=17, are NA=52, NB=52, NX=20, and NY=400? When n=18, are NA=52, NB=53, NX=400, and NY=50? When n=19, are NA=53, NB=53, NX=20, and NY=400? When n=20, are NA=53, NB=54, NX=400, and NY=50? When n=21, are NA=54, NB=54, NX=20, and NY=400? When n=22, are NA=54, NB=55, NX=400, and NY=50? When n=23, NA=55, NB=55, NX=20, and NY=400, or The antisense oligomer or a pharmaceutically acceptable salt thereof or a hydrate thereof, wherein when n=24, NA=55, NB=56, NX=400, and NY=50. [2] Region R1 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 44th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 44th intron. Region R2 is a region consisting of a 600-base sequence from the 3' end to the 5' end of the 44th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 45th exon. Region R3 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 45th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 45th intron. Region R4 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 45th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 46th exon. Region R5 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 46th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 46th intron. Region R6 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 46th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 47th exon. Region R7 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 47th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 47th intron. Region R8 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 47th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 48th exon. Region R9 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 48th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 48th intron. Region R10 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 48th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 49th exon. Region R11 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 49th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 49th intron. Region R12 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 49th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 50th exon. Region R13 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 50th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 50th intron. Region R14 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 50th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 51st exon. Region R15 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 51st exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 51st intron. Region R16 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 51st intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 52nd exon. Region R17 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 52nd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 52nd intron. Region R18 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 52nd intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 53rd exon. Region R19 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 53rd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 53rd intron. Region R20 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 53rd intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 54th exon. Region R21 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 54th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 54th intron. Region R22 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 54th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 55th exon. Region R23 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 55th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 55th intron, or Region R24 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 55th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 56th exon. [1] The antisense oligomers or pharmaceutically acceptable salts thereof, or hydrates thereof. [3] The aforementioned antisense oligomer (a) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389, (b) A nucleotide sequence that hybridizes under stringent conditions with a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389. (c) A nucleotide sequence that has 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389, and has a length within ±15% of the length of the selected nucleotide sequence, or (d) A portion of a base sequence selected from the group consisting of (a), (b), and (c) above. An antisense oligomer according to [1] or [2], or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising a complementary base sequence. [4] The antisense oligomer is an antisense oligomer formed by linking two or more unit oligomers, Each of the aforementioned unit oligomers contains a nucleotide sequence complementary to the nucleotide sequence of any one region selected from the group consisting of regions R1 to R24, or a part thereof, and the nucleotide sequences of each unit oligomer are not continuous or overlapping with each other. The antisense oligomers described in any one of items [1] to [3], or pharmaceutically acceptable salts thereof, or hydrates thereof. [5] The antisense oligomer is an antisense oligomer formed by linking two or more unit oligomers, and each of the unit oligomers is (a) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389, (b) A nucleotide sequence that hybridizes under stringent conditions with a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389. (c) A nucleotide sequence that has 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389, and has a length within ±15% of the length of the selected nucleotide sequence, or (d) A portion of a base sequence selected from the group consisting of (a), (b), and (c) above. An antisense oligomer as described in any one of [1] to [3], or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising a complementary base sequence, wherein the base sequences of each unit oligomer are not continuous or overlapping with each other. [6] The antisense oligomers according to [4] or [5], or pharmaceutically acceptable salts thereof, or hydrates thereof, wherein each of the unit oligomers comprises a base sequence complementary to a continuous base sequence of 5 to 20 base lengths in the region. [7] (1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, or (2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±15% of the length of the selected nucleotide sequence. An antisense oligomer or pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one of items [1] to [3]. [8] Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 130, 131, An antisense oligomer according to any one of items [1] to [3] and [7], or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one base sequence selected from the group consisting of 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232. [9] An antisense oligomer or pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, as described in any one of items [1] to [3] and [7] to [8].

[10] Oligonucleotides, antisense oligomers as described in any one of items [1] to [9], or pharmaceutically acceptable salts thereof, or hydrates thereof.

[11] The antisense oligomer according to

[10] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the sugar moiety and / or phosphate bond moiety of at least one nucleotide constituting the oligonucleotide is modified.

[12] The antisense oligomers described in

[10] or

[11] , or pharmaceutically acceptable salts thereof, or hydrates thereof, wherein the sugar portion of at least one nucleotide constituting the oligonucleotide is a ribose in which the -OH group at the 2' position is substituted with a group selected from the group consisting of -OR, -R, -R'OR, -SH, -SR, -NH2, -NHR, -NR2, -N3, -CN, -F, -Cl, -Br, and -I. (The R represents an alkyl or aryl group, and the R' represents an alkylene group.)

[13] An antisense oligomer according to any one of

[10] to

[12] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the phosphate bond of at least one nucleotide constituting the oligonucleotide is one selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond, and a boranophosphate bond.

[14] Morpholino oligomers, antisense oligomers as described in any one of paragraphs [1] to [9], or pharmaceutically acceptable salts thereof, or hydrates thereof.

[15] The phosphorodiamidate morpholino oligomer, the antisense oligomer described in

[14] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

[16] The antisense oligomers described in

[14] or

[15] , or pharmaceutically acceptable salts thereof, or hydrates thereof, wherein the 5' end is a group of any of the following chemical formulas (1) to (3). [ka]

[17] A suppressor antisense oligomer, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, that inhibits single skipping of any one exon selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA, The suppressor antisense oligomer is (a) Any one of the nucleotide sequences selected from the group consisting of SEQ ID NOs: 370-384, or (b) A nucleotide sequence that has 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 384, and has a length within ±15% of the length of the selected nucleotide sequence, or (c) A portion of the base sequence of the base sequence of (a) or (b) above Contains a complementary base sequence, The suppressor antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

[18] (1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 257 to 275, or (2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 257 to 275, and having a length within ±15% of the length of the selected nucleotide sequence. A suppressor antisense oligomer as described in

[17] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising the above.

[19] A suppressor antisense oligomer according to

[17] or

[18] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one base sequence selected from the group consisting of SEQ ID NOs: 260, 261, and 263.

[20] Oligonucleotides, suppressor antisense oligomers as described in any one of paragraphs

[17] to

[19] , or pharmaceutically acceptable salts thereof, or hydrates thereof. [twenty one] A suppressor antisense oligomer according to

[20] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the sugar moiety and / or phosphate bond moiety of at least one nucleotide constituting the oligonucleotide is modified. [twenty two] The suppressor antisense oligomer according to

[20] or

[21] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the sugar portion of at least one nucleotide constituting the oligonucleotide is a ribose in which the 2'-OH group 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 (The aforementioned R represents alkyl or aryl, and the aforementioned R' represents alkylene.) [twenty three] A suppressor antisense oligomer according to any one of

[20] to

[22] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the phosphate bond of at least one nucleotide constituting the oligonucleotide is one selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond, and a boranophosphate bond. [twenty four] A morpholino oligomer, a suppressor antisense oligomer as described in any one of

[17] to

[19] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [twenty five] A phosphorodiamidate morpholino oligomer, the suppressor antisense oligomer described in

[24] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

[26] A suppressor antisense oligomer as described in

[24] or

[25] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the 5' end is a group of any of the following chemical formulas (1) to (3). [ka]

[27] A pharmaceutical composition comprising an antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, as described in any one of items [1] to

[16] .

[28] The pharmaceutical composition according to

[27] further comprises a suppressor antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, as described in any one of

[17] to

[26] .

[29] A pharmaceutical composition comprising an antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, as described in any one of items [1] to

[16] , and a suppressor antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, as described in any one of items

[17] to

[26] .

[30] (1) The antisense oligomer is an oligomer corresponding to Sequence ID No. 75, and the suppressor antisense oligomer is an oligomer corresponding to Sequence ID No. 260, (2) The antisense oligomer is an oligomer corresponding to SEQ ID NO: 75 and the suppressor antisense oligomer is an oligomer corresponding to SEQ ID NO: 261, or (3) The antisense oligomer is an oligomer corresponding to SEQ ID NO: 75 and the suppressor antisense oligomer is an oligomer corresponding to SEQ ID NO: 263. The pharmaceutical composition described in

[28] or

[29] .

[31] A pharmaceutical composition according to any one of

[27] to

[30] , further comprising a pharmaceutically acceptable carrier.

[32] A pharmaceutical composition according to any one of items

[27] to

[31] for the treatment of muscular dystrophy.

[33] A pharmaceutical composition according to any one of the following paragraphs

[27] to

[32] for administration to a human patient.

[34] A method for treating muscular dystrophy, comprising the step of administering to a patient with muscular dystrophy an antisense oligomer or a pharmaceutically acceptable salt thereof or a hydrate thereof as described in any one of items [1] to

[16] , or a pharmaceutical composition as described in any one of items

[27] to

[33] .

[35] The treatment method according to

[34] , wherein the muscular dystrophy patient is a patient who has a mutation in the dystrophin gene that is subject to exon 45-55 skipping.

[36] The treatment method according to

[34] or

[35] , wherein the patient is a human.

[37] Use of an antisense oligomer or a pharmaceutically acceptable salt thereof or a hydrate thereof, or a pharmaceutical composition, as described in any one of [1] to

[16] , or as described in any one of

[27] to

[33] , in the manufacture of a pharmaceutical for the treatment of muscular dystrophy.

[38] An antisense oligomer according to any one of [1] to

[16] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, for use in the treatment of muscular dystrophy, or a pharmaceutical composition according to any one of

[27] to

[33] .

[39] The antisense oligomer or a pharmaceutically acceptable salt thereof or a hydrate thereof or a pharmaceutical composition according to

[38] , wherein the treatment involves skipping two or more consecutive exons selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA.

[40] In the aforementioned treatment, the patient with muscular dystrophy is a human being, according to the antisense oligomer or pharmaceutically acceptable salt thereof or hydrate thereof or pharmaceutical composition according to

[38] or

[39] .

[41] The process includes a step of inhibiting the splicing silencer sequence, splice site sequence, or branch site sequence of the target pre-mRNA when skipping two or more consecutive exons from the target pre-mRNA. A method for increasing the efficiency of skipping two or more consecutively numbered exons.

[42] The splicing silencer array is heterogeneous The method described in

[41] , which is the recognition sequence for ribonucleoprotein A1 (hnRNPA1).

[43] The method according to

[41] or

[42] , wherein the target pre-mRNA is human dystrophin pre-mRNA.

[44] The method according to any one of

[41] to

[43] , wherein two or more consecutive exons in numerical order are selected from the group consisting of the 45th to 55th exons of human dystrophin pre-mRNA.

[45] The method according to

[44] , wherein skipping of two or more sequentially numbered exons of the target pre-mRNA is performed using an antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, as described in any one of [1] to

[16] .

[46] The method according to

[44] or

[45] , wherein the step of specifically inhibiting the splicing silencer sequence, splice site sequence, or branch site sequence is performed using a suppressor antisense oligomer or a pharmaceutically acceptable salt thereof or a hydrate thereof as described in any one of

[17] to

[26] . 【0017】 Furthermore, the present invention may include the following embodiments.

[47] An antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, which simultaneously skips two or more sequentially numbered exons from a target pre-mRNA, The aforementioned antisense oligomer is (1) A region consisting of a 20-base sequence from the 3' end to the 5' end of any exon of the target pre-mRNA and a 400-base sequence from the 5' end to the 3' end of an intron adjacent to the 3' side of the exon, or (2) A region consisting of a base sequence in the range of 400 bases or 600 bases from the 3' end to the 5' end of any of the introns of the target pre-mRNA, and a base sequence in the range of 50 bases from the 5' end to the 3' end of an exon adjacent to the 3' side of the intron. A sequence containing a nucleotide sequence complementary to the nucleotide sequence of any one region or part thereof selected from the group consisting of the following: The aforementioned antisense oligomers or pharmaceutically acceptable salts thereof or hydrates thereof.

[48] The antisense oligomer described in

[47] , or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the target pre-mRNA is human dystrophin pre-mRNA. [Effects of the Invention] 【0018】 The present invention provides an antisense oligomer that simultaneously skips multiple exons of a target. Another aspect of the present invention provides a pharmaceutical composition for treating patients with various mutations by simultaneously skipping multiple exons of a target pre-mRNA. Another aspect of the present invention provides a suppressor antisense oligomer or a pharmaceutical composition containing said oligomer that suppresses the single skipping of exons of a target pre-mRNA. Another aspect of the present invention makes it possible to simultaneously and efficiently skip exons 45-55 of human dystrophin pre-mRNA. [Brief explanation of the drawing] 【0019】 [Figure 1] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells (human rhabdomyosarcoma cells). [Figure 2] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 3]This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 4] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 5] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 6] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 7] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 8] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 9] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 10] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 11] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 12] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 13] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 14] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 15] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 16] This figure shows the results of RT-PCR investigation into the skipping of exons 45-55 of human dystrophin pre-mRNA in RD cells. [Figure 17] This figure shows the results of RT-PCR investigations into multi-skipping of exons 45-55 and single-skipping of exon 45 of human dystrophin pre-mRNA in RD cells. [Figure 18] This figure shows the results of RT-PCR investigations into multi-skipping of exons 45-55 and single-skipping of exon 45 of human dystrophin pre-mRNA in RD cells. [Figure 19] This figure shows the results of RT-PCR investigations into multi-skipping between exons 45-49 and single-skipping of exon 45 of human dystrophin pre-mRNA in RD cells. [Figure 20] This figure shows the results of examining multi-skipping between exons 45-49 of human dystrophin pre-mRNA in RD cells using RT-PCR, and calculating the total amount of each skipping (total multi-skipping product) for exons 45-46, 45-47, 45-48, and 45-49. All of these skipping processes are thought to have therapeutic effects. [Figure 21] This figure shows the results of RT-PCR investigations into multi-skipping between exons 45-52 and single-skipping of exon 45 of human dystrophin pre-mRNA in RD cells. [Figure 22]This figure shows the results of examining multi-skipping between exons 45-52 of human dystrophin pre-mRNA in RD cells using RT-PCR, and calculating the total amount of skipping between exons 45-47, 45-48, 45-49, 45-50, 45-51, and 45-52 (left) or the total amount of skipping between exons 45-47, 45-48, 45-49, and 45-51 (right). All of the skipping between exons 45-47, 45-48, 45-49, and 45-51 have therapeutic effects. [Modes for carrying out the invention] 【0020】 The present invention will now be described in detail. The following embodiments are illustrative for illustrating the present invention and are not intended to limit the present invention to these embodiments only. The present invention can be implemented in various forms without departing from its spirit. 【0021】 1. Antisense oligomers The present invention relates to an antisense oligomer, a pharmaceutically acceptable salt thereof, or a hydrate thereof, which simultaneously skips two or more sequentially numbered exons from a target pre-mRNA. The aforementioned antisense oligomer is (1) A region consisting of a 20-base sequence from the 3' end to the 5' end of any exon of the target pre-mRNA and a 400-base sequence from the 5' end to the 3' end of an intron adjacent to the 3' side of the exon, or (2) A region consisting of a base sequence in the range of 400 bases or 600 bases from the 3' end to the 5' end of any of the introns of the target pre-mRNA, and a base sequence in the range of 50 bases from the 5' end to the 3' end of an exon adjacent to the 3' side of the intron. A sequence containing a nucleotide sequence complementary to the nucleotide sequence of any one region or part thereof selected from the group consisting of the following: The present invention provides the aforementioned antisense oligomer, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0022】 In this specification, "simultaneously skipping" two or more sequentially numbered exons includes not only cases where the timing of removal of each exon from pre-mRNA is exactly the same, but also cases where each exon is removed sequentially within the period from pre-mRNA to mature mRNA. In other words, "simultaneously skipping" two or more sequentially numbered exons means removing multiple sequentially numbered exons from pre-mRNA. 【0023】 In this specification, "two or more consecutive exons in numerical order" refers to multiple exons in the target pre-mRNA (the total number of exons is denoted as Texon) whose exon numbers increase by 1 each time. Exon number refers to the number assigned to the exon located furthest 5' end of the pre-mRNA, which is designated as the 1st exon, and the exons located sequentially from the 5' end to the 3' end are designated as the 2nd, 3rd, and so on. When skipping two or more consecutive exons in numerical order of a gene, the exon numbers a1,...,a j is a sequence {a j It can be expressed as}. Sequence {a j The general term a of} j It can be expressed by the following formula. [Formula 1] a j =m+(j-1) Here, m is any natural number satisfying 1 ≤ m ≤ (Texon-1), and j is a natural number satisfying 2 ≤ (m+j) ≤ Texon+1. For example, if the target pre-mRNA is human dystrophin pre-mRNA, the texon number is 79. In one embodiment, j is any natural number selected from 1 to 11. In another embodiment, j is 11, j is 10, j is 9, j is 8, j is 7, j is 6, j is 5, j is 4, j is 3, j is 2, and j is 1. 【0024】 In this specification, "gene" includes not only genomic genes but also cDNA, pre-mRNA, and mRNA, and preferably, the gene is pre-mRNA. In this specification, "pre-mRNA" is an RNA molecule containing exons and introns transcribed from a target gene on the genome, and is an mRNA precursor. 【0025】 In one embodiment, the present invention provides an antisense oligomer, a pharmaceutically acceptable salt thereof, or a hydrate thereof that simultaneously skips two or more consecutively numbered exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA (hereinafter, the antisense oligomer, its pharmaceutically acceptable salt, or its hydrate is collectively referred to as "the antisense oligomer of the present invention." The antisense oligomer of the present invention may also refer to the antisense oligomer, its pharmaceutically acceptable salt, or its hydrate individually). 【0026】 Human dystrophin pre-mRNA is an RNA molecule containing exons and introns transcribed from the human dystrophin gene on the genome, and is an mRNA precursor. Those skilled in the art can obtain the nucleotide sequence information of human dystrophin pre-mRNA by analogy with the genomic sequence of the human dystrophin gene (GenBank Accession No. NG_012232.1). 【0027】 In the human genome, the human dystrophin gene is located at locus Xp21.2. The human dystrophin gene is approximately 3.0 Mbp in size, making it the largest known human gene. However, the coding region of the human dystrophin gene is only about 14 kb, and this coding region is distributed within the dystrophin gene as 79 exons (Roberts, RG., et al., Genomics, 16: 536-538 (1993)). The pre-mRNA transcript of the human dystrophin gene undergoes splicing to produce a mature mRNA of approximately 14 kb. The nucleotide sequence of the mature mRNA of the human wild-type dystrophin gene is publicly known (GenBank Accession No. NM_004006). 【0028】 The antisense oligomer of the present invention in the above embodiment is The region Rn (where n is an odd number from 1 to 23) of the human dystrophin pre-mRNA, consisting of a sequence of NX bases from the 3' end to the 5' end of the NA exon and a sequence of NY bases from the 5' end to the 3' end of the NB intron, and The region Rn (where n is an even number from 2 to 24) of human dystrophin pre-mRNA consists of a sequence of NX bases running from the 3' end to the 5' end of the NA intron and a sequence of NY bases running from the 5' end to the 3' end of the NB exon. The target is at least one region selected from the group consisting of regions R1 to R24, as shown by [the formula]. Hereinafter, regions R1 to R24 will be referred to as the "target region of the antisense oligomer of the present invention" or interchangeably as the "target region of the present invention". In this specification, "target" means that the target nucleotide sequence is a nucleotide sequence that is complementary to the nucleotide sequence of the target region or a nucleotide sequence of a part of the target sequence. 【0029】 Furthermore, the antisense oligomer of the present invention in the above embodiment includes a nucleotide sequence complementary to the nucleotide sequence of at least one region selected from the group consisting of target regions R1 to R24 of the present invention, or a nucleotide sequence of a part thereof. 【0030】 Examples of the target region Rn of the present invention in the above embodiment include the following: When n=1, are NA=44, NB=44, NX=20, and NY=400? When n=2, are NA=44, NB=45, NX=600, and NY=50? When n=3, are NA=45, NB=45, NX=20, and NY=400? When n=4, are NA=45, NB=46, NX=400, and NY=50? When n=5, are NA=46, NB=46, NX=20, and NY=400? When n=6, are NA=46, NB=47, NX=400, and NY=50? When n=7, are NA=47, NB=47, NX=20, and NY=400? When n=8, are NA=47, NB=48, NX=400, and NY=50? When n=9, are NA=48, NB=48, NX=20, and NY=400? When n=10, are NA=48, NB=49, NX=400, and NY=50? When n=11, are NA=49, NB=49, NX=20, and NY=400? When n=12, are NA=49, NB=50, NX=400, and NY=50? When n=13, are NA=50, NB=50, NX=20, and NY=400? When n=14, are NA=50, NB=51, NX=400, and NY=50? When n=15, are NA=51, NB=51, NX=20, and NY=400? When n=16, are NA=51, NB=52, NX=400, and NY=50? When n=17, are NA=52, NB=52, NX=20, and NY=400? When n=18, are NA=52, NB=53, NX=400, and NY=50? When n=19, are NA=53, NB=53, NX=20, and NY=400? When n=20, are NA=53, NB=54, NX=400, and NY=50? When n=21, are NA=54, NB=54, NX=20, and NY=400? When n=22, are NA=54, NB=55, NX=400, and NY=50? When n=23, NA=55, NB=55, NX=20, and NY=400, or When n=24, NA=55, NB=56, NX=400, and NY=50. 【0031】 In another embodiment, examples of the target region Rn of the present invention include, but are not limited to, the following. When n=1, are NA=44, NB=44, NX=20, and NY=400? When n=2, are NA=44, NB=45, NX=600, and NY=19? When n=3, are NA=45, NB=45, NX=20, and NY=400? When n=4, are NA=45, NB=46, NX=400, and NY=50? When n=5, are NA=46, NB=46, NX=20, and NY=400? When n=6, are NA=46, NB=47, NX=400, and NY=37? When n=7, are NA=47, NB=47, NX=20, and NY=400? When n=8, are NA=47, NB=48, NX=400, and NY=19? When n=9, are NA=48, NB=48, NX=20, and NY=400? When n=10, are NA=48, NB=49, NX=400, and NY=42? When n=11, are NA=49, NB=49, NX=20, and NY=400? When n=12, are NA=49, NB=50, NX=400, and NY=44? When n=13, are NA=50, NB=50, NX=20, and NY=400? When n=14, are NA=50, NB=51, NX=400, and NY=25? When n=15, are NA=51, NB=51, NX=20, and NY=400? When n=16, are NA=51, NB=52, NX=400, and NY=24? When n=17, are NA=52, NB=52, NX=20, and NY=400? When n=18, are NA=52, NB=53, NX=400, and NY=34? When n=19, are NA=53, NB=53, NX=20, and NY=400? When n=20, are NA=53, NB=54, NX=400, and NY=43? When n=21, are NA=54, NB=54, NX=20, and NY=400? When n=22, are NA=54, NB=55, NX=400, and NY=25? When n=23, NA=55, NB=55, NX=20, and NY=400, or When n=24, NA=55, NB=56, NX=400, and NY=50. 【0032】 Examples of the target region Rn of the present invention, R1 to R24, may be expressed as follows in other embodiments, but are not limited thereto. For example, in one embodiment, region R1 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 44th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 44th intron. Region R2 is a region consisting of a 600-base sequence from the 3' end to the 5' end of the 44th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 45th exon. Region R3 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 45th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 45th intron. Region R4 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 45th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 46th exon. Region R5 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 46th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 46th intron. Region R6 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 46th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 47th exon. Region R7 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 47th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 47th intron. Region R8 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 47th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 48th exon. Region R9 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 48th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 48th intron. Region R10 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 48th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 49th exon. Region R11 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 49th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 49th intron. Region R12 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 49th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 50th exon. Region R13 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 50th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 50th intron. Region R14 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 50th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 51st exon. Region R15 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 51st exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 51st intron. Region R16 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 51st intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 52nd exon. Region R17 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 52nd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 52nd intron. Region R18 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 52nd intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 53rd exon. Region R19 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 53rd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 53rd intron. Region R20 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 53rd intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 54th exon. Region R21 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 54th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 54th intron. Region R22 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 54th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 55th exon. Region R23 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 55th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 55th intron, or Region R24 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 55th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 56th exon. 【0033】 In another embodiment, the target region Rn of the present invention, R1 to R24, may be expressed as follows in the above embodiment, but is not limited thereto. For example, in one embodiment, region R1 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 44th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 44th intron. Region R2 is a region consisting of a 600-base sequence from the 3' end to the 5' end of the 44th intron of the human dystrophin pre-mRNA and a 19-base sequence from the 5' end to the 3' end of the 45th exon. Region R3 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 45th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 45th intron. Region R4 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 45th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 46th exon. Region R5 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 46th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 46th intron. Region R6 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 46th intron of the human dystrophin pre-mRNA and a 37-base sequence from the 5' end to the 3' end of the 47th exon. Region R7 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 47th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 47th intron. Region R8 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 47th intron of the human dystrophin pre-mRNA and a 19-base sequence from the 5' end to the 3' end of the 48th exon. Region R9 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 48th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 48th intron. Region R10 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 48th intron of the human dystrophin pre-mRNA and a 42-base sequence from the 5' end to the 3' end of the 49th exon. Region R11 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 49th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 49th intron. Region R12 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 49th intron of the human dystrophin pre-mRNA and a 44-base sequence from the 5' end to the 3' end of the 50th exon. Region R13 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 50th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 50th intron. Region R14 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 50th intron of the human dystrophin pre-mRNA and a 25-base sequence from the 5' end to the 3' end of the 51st exon. Region R15 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 51st exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 51st intron. Region R16 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 51st intron of the human dystrophin pre-mRNA and a 24-base sequence from the 5' end to the 3' end of the 52nd exon. Region R17 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 52nd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 52nd intron. Region R18 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 52nd intron of the human dystrophin pre-mRNA and a 34-base sequence from the 5' end to the 3' end of the 53rd exon. Region R19 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 53rd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 53rd intron. Region R20 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 53rd intron of the human dystrophin pre-mRNA and a 43-base sequence from the 5' end to the 3' end of the 54th exon. Region R21 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 54th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 54th intron. Region R22 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 54th intron of the human dystrophin pre-mRNA and a 25-base sequence from the 5' end to the 3' end of the 55th exon. Region R23 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 55th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 55th intron, or Region R24 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 55th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 56th exon. 【0034】 Furthermore, while R1 to R24, which are examples of the target region Rn of the present invention, may be expressed as follows in other embodiments, they are not limited to these. (A1) Area R1 Region R1 is defined as the region ranging from -20 bases to +400 bases, where the boundary between the 3' end of exon 44 and the 5' end of intron 44 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is designated as "+" and the base sequence region 3' (downstream) is designated as "-" (minus). In this case, the region ranging from -20 bases to -1 base belongs to exon 44, and the region ranging from +1 base to +400 base belongs to intron 44. (A2) Area R2 Region R2 is defined as the region ranging from -600 bases to +50 bases, where the boundary between the 3' end of intron 44 and the 5' end of exon 45 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -600 bases to -1 base belongs to intron 44, and the region ranging from +1 base to +50 base belongs to exon 45. (A3) Area R3 Region R3 is defined as the region ranging from -20 bases to +400 bases, where the boundary between the 3' end of exon 45 and the 5' end of intron 45 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -20 bases to -1 base belongs to exon 45, and the region ranging from +1 base to +400 base belongs to intron 45. (A4) Area R4 Region R4 is defined as the region ranging from -400 bases to +50 bases, where the boundary between the 3' end of intron 45 and the 5' end of exon 46 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 45, and the region ranging from +1 base to +50 base belongs to exon 46. (A5) Area R5 Region R5 is defined as the region ranging from -20 bases to +400 bases, where the boundary between the 3' end of exon 46 and the 5' end of intron 46 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -20 bases to -1 base belongs to exon 46, and the region ranging from +1 base to +400 base belongs to intron 46. (A6) Area R6 Region R6 is defined as the region ranging from -400 bases to +50 bases, where the boundary between the 3' end of intron 46 and the 5' end of exon 47 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 46, and the region ranging from +1 base to +50 base belongs to exon 47. (A7) Area R7 Region R7 is the region represented by the range of -20 bases to +400 bases, where the boundary between the 3' end of exon 47 and the 5' end of intron 47 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by the range of -20 bases to -1 base belongs to exon 47, and the region represented by the range of +1 base to +400 base belongs to intron 47. (A8) Area R8 Region R8 is the region represented by a range of -400 bases to +50 bases, where the boundary between the 3' end of intron 47 and the 5' end of exon 48 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by -400 bases to -1 base belongs to intron 47, and the region represented by +1 base to +50 base belongs to exon 48. (A9) Area R9 Region R9 is the region represented by the range of -20 bases to +400 bases, where the boundary between the 3' end of exon 48 and the 5' end of intron 48 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by the range of -20 bases to -1 base belongs to exon 48, and the region represented by the range of +1 base to +400 bases belongs to intron 48. (A10) Area R10 Region R10 is defined as the region ranging from -400 bases to +50 bases, where the boundary between the 3' end of intron 48 and the 5' end of exon 49 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 48, and the region ranging from +1 base to +50 base belongs to exon 49. (A11) Area R11 Region R11 is the region represented by the range of -20 bases to +400 bases, where the boundary between the 3' end of exon 49 and the 5' end of intron 49 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by the range of -20 bases to -1 base belongs to exon 49, and the region represented by the range of +1 base to +400 bases belongs to intron 49. (A12) Area R12 Region R12 is the region represented by a range of -400 bases to +50 bases, where the boundary between the 3' end of intron 49 and the 5' end of exon 50 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by -400 bases to -1 base belongs to intron 49, and the region represented by +1 base to +50 base belongs to exon 50. (A13) Area R13 Region R13 is the region represented by the range of -20 bases to +400 bases, where the boundary between the 3' end of exon 50 and the 5' end of intron 50 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), and the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region represented by the range of -20 bases to -1 base belongs to exon 50, and the region represented by the range of +1 base to +400 base belongs to intron 50. (A14) Area R14 Region R14 is the region represented in the range of -400 bases to +50 bases, where the boundary between the 3' end of intron 50 and the 5' end of exon 51 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus) and the base sequence region 3' (downstream) is indicated by "+". In this case, the region represented in the range of -400 bases to -1 base belongs to intron 50, and the region represented in the range of +1 base to +50 base belongs to exon 51. (A15) Area R15 Region R15 is the region represented by the range of -20 bases to +400 bases, where the boundary between the 3' end of exon 51 and the 5' end of intron 51 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), and the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region represented by the range of -20 bases to -1 base belongs to exon 51, and the region represented by the range of +1 base to +400 base belongs to intron 51. (A16) Area R16 Region R16 is defined as the region ranging from -400 bases to +50 bases, where the boundary between the 3' end of intron 51 and the 5' end of exon 52 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus) and the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 51, and the region ranging from +1 base to +50 base belongs to exon 52. (A17) Area R17 Region R17 is the region represented by the range of -20 bases to +400 bases, where the boundary between the 3' end of exon 52 and the 5' end of intron 52 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus) and the base sequence region 3' (downstream) is indicated by "+". In this case, the region represented by -20 bases to -1 base belongs to exon 52, and the region represented by +1 base to +400 base belongs to intron 52. (A18) Area R18 Region R18 is defined as the region ranging from -400 bases to +50 bases, where the boundary between the 3' end of intron 52 and the 5' end of exon 53 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus) and the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 52, and the region ranging from +1 base to +50 base belongs to exon 53. (A19) Area R19 Region R19 is defined as the region ranging from -20 bases to +400 bases, where the boundary between the 3' end of exon 53 and the 5' end of intron 53 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus) and the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -20 bases to -1 base belongs to exon 53, and the region ranging from +1 base to +400 base belongs to intron 53. (A20) Area R20 Region R20 is defined as the region ranging from -400 bases to +50 bases, where the boundary between the 3' end of intron 53 and the 5' end of exon 54 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), while the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 53, and the region ranging from +1 base to +50 base belongs to exon 54. (A21) Area R21 Region R21 is defined as the region ranging from -20 bases to +400 bases, where the boundary between the 3' end of exon 54 and the 5' end of intron 54 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), while the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region ranging from -20 bases to -1 base belongs to exon 54, and the region ranging from +1 base to +400 base belongs to intron 54. (A22) Area R22 Region R22 is the region represented by a range of -400 bases to +50 bases, where the boundary between the 3' end of intron 54 and the 5' end of exon 55 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by -400 bases to -1 base belongs to intron 54, and the region represented by +1 base to +50 base belongs to exon 55. (A23) Area R23 Region R23 is defined as the region ranging from -20 bases to +400 bases, where the boundary between the 3' end of exon 55 and the 5' end of intron 55 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus) and the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -20 bases to -1 base belongs to exon 55, and the region ranging from +1 base to +400 base belongs to intron 55. (A24) Area R24 Region R24 is the region represented by a range of -400 bases to +50 bases, where the boundary between the 3' end of intron 55 and the 5' end of exon 56 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by -400 bases to -1 base belongs to intron 55, and the region represented by +1 base to +50 base belongs to exon 56. 【0035】 In another embodiment, examples of the target region Rn of the present invention, R2, R6, R8, R10, R12, R14, R16, R18, R20, and R22, may also be represented as follows in the above embodiment, but are not limited thereto. (A2) Area R2 Region R2 is defined as the region ranging from -600 bases to +19 bases, where the boundary between the 3' end of intron 44 and the 5' end of exon 45 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus) and the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -600 bases to -1 base belongs to intron 44, and the region ranging from +1 base to +19 base belongs to exon 45. (A6) Area R6 Region R6 is defined as the region ranging from -400 bases to +37 bases, where the boundary between the 3' end of intron 46 and the 5' end of exon 47 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is denoted as "-" (minus), while the base sequence region 3' (downstream) is denoted as "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 46, and the region ranging from +1 base to +37 base belongs to exon 47. (A8) Area R8 Region R8 is the region represented by a base sequence of -400 to +19 bases, where the boundary between the 3' end of intron 47 and the 5' end of exon 48 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by a base sequence of -400 to -1 bases belongs to intron 47, and the region represented by a base sequence of +1 to +19 bases belongs to exon 48. (A10) Area R10 Region R10 is the region represented by a base sequence of -400 to +42 bases, where the boundary between the 3' end of intron 48 and the 5' end of exon 49 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by a base sequence of -400 to -1 base belongs to intron 48, and the region represented by a base sequence of +1 to +42 bases belongs to exon 49. (A12) Area R12 Region R12 is the region represented by a base sequence of -400 to +44 bases, where the boundary between the 3' end of intron 49 and the 5' end of exon 50 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by a base sequence of -400 to -1 base belongs to intron 49, and the region represented by a base sequence of +1 to +44 bases belongs to exon 50. (A14) Area R14 Region R14 is the region represented by a base sequence of -400 to +25 bases, where the boundary between the 3' end of intron 50 and the 5' end of exon 51 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), and the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region represented by a base sequence of -400 to -1 base belongs to intron 50, and the region represented by a base sequence of +1 to +25 bases belongs to exon 51. (A16) Area R16 Region R16 is the region represented by a base sequence of -400 to +24 bases, where the boundary between the 3' end of intron 51 and the 5' end of exon 52 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is represented by "-" (minus), and the base sequence region 3' (downstream) from the base point is represented by "+". In this case, the region represented by a base sequence of -400 to -1 base belongs to intron 51, and the region represented by a base sequence of +1 to +24 bases belongs to exon 52. (A18) Area R18 Region R18 is the region represented by a range of -400 bases to +34 bases, where the boundary between the 3' end of intron 52 and the 5' end of exon 53 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), and the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region represented by -400 bases to -1 base belongs to intron 52, and the region represented by +1 base to +34 base belongs to exon 53. (A20) Area R20 Region R20 is defined as the region ranging from -400 bases to +43 bases, where the boundary between the 3' end of intron 53 and the 5' end of exon 54 is designated as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), while the base sequence region 3' (downstream) is indicated by "+". In this case, the region ranging from -400 bases to -1 base belongs to intron 53, and the region ranging from +1 base to +43 base belongs to exon 54. (A22) Area R22 Region R22 is the region represented by a range of -400 bases to +25 bases, where the boundary between the 3' end of intron 54 and the 5' end of exon 55 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by "-" (minus), and the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region represented by -400 bases to -1 base belongs to intron 54, and the region represented by +1 base to +25 base belongs to exon 55. 【0036】 Specific examples of the nucleotide sequences of regions R1 to R24 are given below. In this specification, thymine "T" and uracil "U" are interchangeable, and whether it is "T" or "U" does not essentially affect the exon skipping activity of the antisense oligomer of the present invention. Therefore, in the nucleotide sequences shown herein, cases where "T" is "U" are also included and will be indicated by the same sequence number. In the table below, even in the nucleotide sequence of pre-mRNA, "U" may be represented as "T", but those skilled in the art can appropriately replace "T" with "U" to understand the RNA sequence. 【0037】 [Table 1] TIFF2026095457000004.tif255167TIFF2026095457000005.tif255164TIFF2026095457000006.tif255164TIFF2026095457000007.tif150170 【0038】 In another embodiment, specific examples of the base sequences of regions R2, R6, R8, R10, R12, R14, R16, R18, R20, and R22 are also given below. 【0039】 [Table 2] TIFF2026095457000009.tif255163 【0040】 In yet another embodiment, the antisense oligomer of the present invention is The Rn region (where n is an odd number from 1 to 23) consisting of a sequence of NY bases, from the 5' end to the 3' end of the NB intron of human dystrophin pre-mRNA, and The Rn region (where n is an even number from 2 to 24) consists of an NX base sequence running from the 3' end to the 5' end of the NA intron of human dystrophin pre-mRNA. The target is at least one region selected from the group consisting of regions R1 to R24, as indicated by [the symbol]. 【0041】 In the above embodiment, examples of the target region Rn of the present invention include, but are not limited to, the following. When n=1, are NB=44 and NY=400? When n=2, are NA=44 and NX=600? When n=3, are NB=45 and NY=400? When n=4, are NA=45 and NX=400? When n=5, are NB=46 and NY=400? When n=6, are NA=46 and NX=400? When n=7, are NB=47 and NY=400? When n=8, are NA=47 and NX=400? When n=9, are NB=48 and NY=400? When n=10, are NA=48 and NX=400? When n=11, are NB=49 and NY=400? When n=12, are NA=49 and NX=400? When n=13, are NB=50 and NY=400? When n=14, are NA=50 and NX=400? When n=15, are NB=51 and NY=400? When n=16, are NA=51 and NX=400? When n=17, are NB=52 and NY=400? When n=18, are NA=52 and NX=400? When n=19, are NB=53 and NY=400? When n=20, are NA=53 and NX=400? When n=21, are NB=54 and NY=400? When n=22, are NA=54 and NX=400? When n=23, NB=55 and NY=400, or When n=24, NA=55 and NX=400. 【0042】 Examples of the target region Rn of the present invention according to the above embodiment, R1 to R24, may be expressed as follows in another embodiment, but are not limited thereto. For example, in one embodiment, region R1 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 44th intron of the human dystrophin pre-mRNA. Region R2 is a region consisting of a 600-base sequence from the 3' end to the 5' end of the 44th intron of the human dystrophin pre-mRNA. Region R3 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 45th intron of the human dystrophin pre-mRNA. Region R4 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 45th intron of the human dystrophin pre-mRNA. Region R5 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 46th intron of the human dystrophin pre-mRNA. Region R6 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 46th intron of the human dystrophin pre-mRNA. Region R7 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 47th intron of the human dystrophin pre-mRNA. Region R8 is a 400-base region in the direction from the 3' end to the 5' end of the 47th intron of the human dystrophin pre-mRNA. Region R9 is a region consisting of a 400-base sequence extending from the 5' end to the 3' end of the 48th intron of the human dystrophin pre-mRNA. Region R10 is a region consisting of a 400-base sequence extending from the 3' end to the 5' end of the 48th intron of the human dystrophin pre-mRNA. Region R11 is a region consisting of a 400-base sequence extending from the 5' end to the 3' end of the 49th intron of the human dystrophin pre-mRNA. Region R12 is a region consisting of a 400-base sequence extending from the 3' end to the 5' end of the 49th intron of the human dystrophin pre-mRNA. Region R13 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 50th intron of the human dystrophin pre-mRNA. Region R14 is a region consisting of a 400-base sequence extending from the 3' end to the 5' end of the 50th intron of the human dystrophin pre-mRNA. Region R15 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 51st intron of the human dystrophin pre-mRNA. Region R16 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 51st intron of the human dystrophin pre-mRNA. Region R17 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 52nd intron of the human dystrophin pre-mRNA. Region R18 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 52nd intron of the human dystrophin pre-mRNA. Region R19 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 53rd intron of the human dystrophin pre-mRNA. Region R20 is a region consisting of a 400-base sequence extending from the 3' end to the 5' end of the 53rd intron of the human dystrophin pre-mRNA. Region R21 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 54th intron of the human dystrophin pre-mRNA. Region R22 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 54th intron of the human dystrophin pre-mRNA. Region R23 is a region consisting of a 400-base sequence from the 5' end to the 3' end of the 55th intron of the human dystrophin pre-mRNA, or Region R24 is a region consisting of a 400-base sequence extending from the 3' end to the 5' end of the 55th intron of the human dystrophin pre-mRNA. 【0043】 Furthermore, while R1 to R24, which are examples of the target region Rn of the present invention according to the above embodiment, may be expressed as follows in other embodiments, they are not limited to these. (A1) Area R1 Region R1 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 44 and the 5' end of intron 44 is taken as base point 0, and the base sequence region 3' (downstream) from the base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 44. (A2) Area R2 Region R2 is the region represented by -600 bases to -1 base, where the base sequence region 5' upstream from the base point 0 (5' end of intron 44) is indicated by a "-" (minus sign). In this case, the region represented by -600 bases to -1 base belongs to intron 44. (A3) Area R3 Region R3 is defined as the region between +1 bases and +400 bases, where the boundary between the 3' end of exon 45 and the 5' end of intron 45 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by a "+". In this case, the region between +1 bases and +400 bases belongs to intron 45. (A4) Area R4 Region R4 is the region represented by -400 bases to -1 base, where the base sequence region 5' upstream from the base point 0 (5' end of intron 45) is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 45. (A5) Area R5 Region R5 is defined as the region between +1 bases and +400 bases when the base sequence region 3' downstream from the base point 0 (the boundary between the 3' end of exon 46 and the 5' end of intron 46) is indicated by a "+". In this case, the region between +1 bases and +400 bases belongs to intron 46. (A6) Area R6 Region R6 is the region represented by -400 bases to -1 base, where the base sequence region 5' upstream from the base point 0 (5' end of intron 46) is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 46. (A7) Area R7 Region R7 is defined as the region between +1 bases and +400 bases when the base sequence region 3' downstream from the base point 0 (the boundary between the 3' end of exon 47 and the 5' end of intron 47) is indicated by a "+". In this case, the region between +1 bases and +400 bases belongs to intron 47. (A8) Area R8 Region R8 is the region represented by -400 bases to -1 base, where the base sequence region 5' upstream from the base point 0 (5' end of intron 47) is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 47. (A9) Area R9 Region R9 is defined as the region between +1 bases and +400 bases, where the boundary between the 3' end of exon 48 and the 5' end of intron 48 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by a "+". In this case, the region between +1 bases and +400 bases belongs to intron 48. (A10) Area R10 Region R10 is the region represented by -400 bases to -1 base, where the base sequence region 5' upstream from the base point 0 (5' end of intron 48 and 5' end of exon 49) is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 48. (A11) Area R11 Region R11 is the region represented by the range of +1 to +400 bases, where the boundary between the 3' end of exon 49 and the 5' end of intron 49 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 49. (A12) Area R12 Region R12 is the region represented by -400 bases to -1 base, where the base sequence region 5' upstream of the base point 0 (5' end of intron 49) is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 49. (A13) Area R13 Region R13 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 50 and the 5' end of intron 50 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 50. (A14) Area R14 Region R14 is the region represented by a base sequence of -400 bases to -1 base, where the boundary between the 3' end of intron 50 and the 5' end of exon 51 is taken as base point 0, and the base sequence region 5' (upstream) from base point is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 50. (A15) Area R15 Region R15 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 51 and the 5' end of intron 51 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 51. (A16) Area R16 Region R16 is the region represented by a base sequence of -400 bases to -1 base, where the boundary between the 3' end of intron 51 and the 5' end of exon 52 is taken as base point 0, and the base sequence region 5' upstream from the base point is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 51. (A17) Area R17 Region R17 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 52 and the 5' end of intron 52 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 52. (A18) Area R18 Region R18 is the region represented by a base sequence of -400 bases to -1 base, where the boundary between the 3' end of intron 52 and the 5' end of exon 53 is taken as base point 0, and the base sequence region 5' upstream from the base point is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 52. (A19) Area R19 Region R19 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 53 and the 5' end of intron 53 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 53. (A20) Area R20 Region R20 is the region represented by -400 bases to -1 base, where the base sequence region 5' upstream of the base point 0 (5' end of intron 53 and 5' end of exon 54) is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 53. (A21) Area R21 Region R21 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 54 and the 5' end of intron 54 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 54. (A22) Area R22 Region R22 is the region represented by a base sequence of -400 bases to -1 base, where the boundary between the 3' end of intron 54 and the 5' end of exon 55 is taken as base point 0, and the base sequence region 5' (upstream) from the base point is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 54. (A23) Area R23 Region R23 is the region represented by +1 to +400 bases, where the boundary between the 3' end of exon 55 and the 5' end of intron 55 is taken as base point 0, and the base sequence region 3' (downstream) from base point is indicated by "+". In this case, the region represented by +1 to +400 bases belongs to intron 55. (A24) Area R24 Region R24 is the region represented by a base sequence of -400 bases to -1 base, where the boundary between the 3' end of intron 55 and the 5' end of exon 56 is taken as base point 0, and the base sequence region 5' upstream from the base point is indicated by a "-" (minus sign). In this case, the region represented by -400 bases to -1 base belongs to intron 55. 【0044】 In the above embodiment, specific examples of the base sequences of regions R1 to R24 are as follows. 【0045】 [Table 3] TIFF2026095457000011.tif255164TIFF2026095457000012.tif255164TIFF2026095457000013.tif255164TIFF2026095457000014.tif85170 【0046】 Furthermore, the target regions R1 to R24 of the antisense oligomer of the present invention include not only the wild type (e.g., the regions shown in SEQ ID NOs. 233 to 256, 341 to 369, and 385 to 389) but also mutant forms of human dystrophin pre-mRNA. Specifically, such mutant forms have one base sequence selected from the group consisting of the following base sequences (B0) and (B1) to (B16). (B0) A nucleotide sequence that hybridizes under stringent conditions with a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389; (B1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±15% of the length of the selected nucleotide sequence. (B2) A nucleotide sequence that has 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and whose length is within ±14% of the length of the selected nucleotide sequence. (B3) A nucleotide sequence that has 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and whose length is within ±13% of the length of the selected nucleotide sequence. (B4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±12% of the length of the selected nucleotide sequence. (B5) A nucleotide sequence that has 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and whose length is within ±11% of the length of the selected nucleotide sequence. (B6) A nucleotide sequence that has 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±10% of the length of the selected nucleotide sequence. (B7) A nucleotide sequence that has 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±9% of the length of the selected nucleotide sequence. (B8) A nucleotide sequence that has 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±8% of the length of the selected nucleotide sequence. (B9) A nucleotide sequence that has 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±7% of the length of the selected nucleotide sequence. (B10) A nucleotide sequence that has 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±6% of the length of the selected nucleotide sequence. (B11) A nucleotide sequence that has 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±5% of the length of the selected nucleotide sequence. (B12) A nucleotide sequence that has 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±4% of the length of the selected nucleotide sequence. (B13) A nucleotide sequence that has 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±3% of the length of the selected nucleotide sequence. (B14) A nucleotide sequence that has 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±2% of the length of the selected nucleotide sequence. (B15) A nucleotide sequence that has 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and has a length within ±1% of the length of the selected nucleotide sequence. (B16) A nucleotide sequence that has 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and whose length is within ±0.5% of the length of the selected nucleotide sequence. 【0047】 In this specification, "nucleotide sequences that hybridize under stringent conditions" refers to nucleotide sequences obtained by using colony hybridization, plaque hybridization, or Southern hybridization, for example, by using all or part of a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389 as a probe. Hybridization methods can be those described in, for example, "Sambrook & Russel, Molecular Cloning: A Laboratory Manual Vol. 3, Cold Spring Harbor Laboratory Press, 2001" and "Ausubel, Current Protocols in Molecular Biology, John Wiley & Sons, 1987-1997." 【0048】 In this specification, "complementary base sequence" is not limited to base sequences that form Watson-Crick pairs with the target base sequence, but also includes base sequences that form wobble base pairs. Here, Watson-Crick pairs refer to base pairs in which hydrogen bonds are formed between adenine-thymine, adenine-uracil, and guanine-cytosine, while wobble base pairs refer to base pairs in which hydrogen bonds are formed between guanine-uracil, inosine-uracil, inosine-adenine, and inosine-cytosine. Furthermore, "complementary base sequences" do not necessarily have 100% complementarity with the target base sequence. For example, they may contain one, two, three, four, or five non-complementary bases compared to the target base sequence, or they may be base sequences that are one, two, three, four, or five bases shorter than the target base sequence. 【0049】 In this specification, “stringent conditions” may refer to any of the low-stringent, medium-stringent, or high-stringent conditions. “Low-stringent conditions” are, for example, 5×SSC, 5×Denhardt solution, 0.5% SDS, 50% formamide, and 32°C. “Medium-stringent conditions” are, for example, 5×SSC, 5×Denhardt solution, 0.5% SDS, 50% formamide, and 42°C, or 5×SSC, 1% SDS, 50mM Tris-HCl (pH 7.5), 50% formamide, and 42°C. “High-stringent conditions” are, for example, 5×SSC, 5×Denhardt solution, 0.5% SDS, 50% formamide, and 50°C, or 0.2×SSC, 0.1% SDS, and 65°C. Under these conditions, it can be expected that base sequences with high sequence identity can be efficiently obtained by increasing the temperature. However, several factors such as temperature, probe concentration, probe length, ionic strength, time, and salt concentration can be considered to affect the stringency of hybridization, and those skilled in the art can achieve similar stringency by appropriately selecting these factors. When using a commercially available kit for hybridization, for example, the AlkPhos Direct Labelling and Detection System (GE Healthcare) can be used. In this case, following the protocol provided with the kit, the labeled probe is incubated overnight, and then the membrane is washed with a primary washing buffer containing 0.1% (w / v) SDS at 55°C before hybridization can be detected. Alternatively, when preparing a probe based on all or part of a nucleotide sequence complementary to one of the nucleotide sequences selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, if the probe is labeled with digoxigenin (DIG) using a commercially available reagent (e.g., PCR labeling mix (Roche Diagnostics)), hybridization can be detected using a DIG nucleic acid detection kit (Roche Diagnostics) or the like. 【0050】 The identity of nucleotide sequences can be determined using the BLAST (Basic Local Alignment Search Tool) algorithm by Carlin and Arthur (Proc. Natl. Acad. Sci. USA 872264-2268, 1990; Proc Natl Acad Sci USA 90: 5873, 1993). Programs called BLASTN and BLASTX, based on the BLAST algorithm, have been developed (Altschul SF, et al: J Mol Biol 215: 403, 1990). When analyzing nucleotide sequences using BLASTN, the parameters should be, for example, score=100 and wordlength=12. When using the BLAST and Gapped BLAST programs, use the default parameters of each program. 【0051】 The antisense oligomer of the present invention contains a nucleotide sequence complementary to the nucleotide sequence of at least one region selected from the group consisting of target regions R1 to R24 of the present invention, or a nucleotide sequence of a part thereof. "A part thereof" means a region excluding the entire length of at least one region selected from the group consisting of target regions R1 to R24, that is, a subregion of at least one region selected from the group consisting of target regions R1 to R24. The lengths of the subregions are 10-60 nucleotides, 10-55 nucleotides, 10-50 nucleotides, 10-45 nucleotides, 10-40 nucleotides, 10-35 nucleotides, 10-30 nucleotides, 10-25 nucleotides, 15-60 nucleotides, 15-55 nucleotides, 15-50 nucleotides, 15-45 nucleotides, 15-40 nucleotides, 15-35 nucleotides, 15-30 nucleotides, 15-25 nucleotides, 16-60 nucleotides, 16-55 nucleotides, 16-50 nucleotides, 16-45 nucleotides, 16-40 nucleotides, and 16-3 5 nucleotides long, 16-30 nucleotides long, 16-25 nucleotides long, 17-60 nucleotides long, 17-55 nucleotides long, 17-45 nucleotides long, 17-40 nucleotides long, 17-35 nucleotides long, 17-30 nucleotides long, 17-25 nucleotides long, 18-60 nucleotides long, 18-55 nucleotides long, 18-50 nucleotides long, 18-45 nucleotides long, 18-40 nucleotides long, 18-35 nucleotides long, 18-30 nucleotides long, 18-25 nucleotides long, 19-60 nucleotides long, 19-55 nucleotides long, 19-50 nucleotides long, 19-45 nucleotides long, 19-40 nucleotides long, 19-35 nucleotides long, 19-30 nucleotides long, 19-25 nucleotides long, 20-60 nucleotides long, 20-55 nucleotides long, 20-50 nucleotides long, 20-45 nucleotides long, 20-40 nucleotides long, 20-35 nucleotides long, 20-30 nucleotides long, 20-25 nucleotides long, 15-30 nucleotides long, 15-29 nucleotides long, 15-28 nucleotides long, 15-27 nucleotides long, 15-26 nucleotides long, 15-25 nucleotides long, 15-24 nucleotides long, 15-23 nucleotides long, 15-22 nucleotides long, 15-21 nucleotides long, 15-2 0 nucleotides, 15-19 nucleotides, 15-18 nucleotides, 16-30 nucleotides, 16-29 nucleotides, 16-28 nucleotides, 16-27 nucleotides, 16-26 nucleotides, 16-25 nucleotides, 16-24 nucleotides, 16-23 nucleotides, 16-22 nucleotides, 16-21 nucleotides, 16-20 nucleotides, 16-19 nucleotides, 16-18 nucleotides, 17-30 nucleotides, 17-29 nucleotides, 17-28 nucleotides, 17-27 nucleotides, 17-26 nucleotides, 17-25 nucleotides, 17-24 nucleotides,17-23 nucleotide lengths, 17-22 nucleotide lengths, 17-21 nucleotide lengths, 17-20 nucleotide lengths, 17-19 nucleotide lengths, 17-18 nucleotide lengths, 18-30 nucleotide lengths, 18-29 nucleotide lengths, 18-28 nucleotide lengths, 18-27 nucleotide lengths, 18-26 nucleotide lengths, 18-25 nucleotide lengths, 18-24 nucleotide lengths, 18-23 nucleotide lengths, 18-22 nucleotide lengths, 18-21 nucleotide lengths, 18-20 nucleotide lengths, 18-19 nucleotide lengths, 19-30 nucleotide lengths, 19-29 nucleotide lengths, 19-28 nucleotide lengths, 19-27 nucleotide lengths, 19-26 nucleotide lengths, 19-25 nucleotide lengths, 19-24 nucleotide lengths, 19-23 nucleotide lengths, 19-22 nucleotide lengths, 19-21 salt Base length, 19-20 nucleotides length, 20-30 nucleotides length, 20-29 nucleotides length, 20-28 nucleotides length, 20-27 nucleotides length, 20-26 nucleotides length, 20-25 nucleotides length, 20-24 nucleotides length, 20-23 nucleotides length, 20-22 nucleotides length, 20-21 nucleotides length, 5-25 nucleotides length, 5-24 nucleotides length, 5-23 nucleotides length, 5-22 nucleotides length, 5-21 nucleotides length, 5-20 nucleotides length, 5-19 nucleotides length, 5-18 nucleotides length, 5-17 nucleotides length, 5-16 nucleotides length, 5-15 nucleotides length, 5-14 nucleotides length, 5-13 nucleotides length, 5-12 nucleotides length, 7-25 nucleotides length, 7-24 nucleotides length, 7-23 nucleotides length, 7-22 nucleotides length, 7-21 Base length, 7-20 base lengths, 7-19 base lengths, 7-18 base lengths, 7-17 base lengths, 7-16 base lengths, 7-15 base lengths, 7-14 base lengths, 7-13 base lengths, 7-12 base lengths, 9-25 base lengths, 9-24 base lengths, 9-23 base lengths, 9-22 base lengths, 9-21 base lengths, 9-20 base lengths, 9-19 base lengths, 9-18 base lengths, 9-17 base lengths, 9-16 base lengths, 9-15 base lengths, 9-14 base lengths, 9-13 base lengths, 9-12 base lengths, 10-25 base lengths, 10-24 base lengths, 10-23 base lengths, 10-22 base lengths, 10-21 base lengths, 10-20 base lengths, 10-19 base lengths , 10-18 nucleotides long, 10-17 nucleotides long, 10-16 nucleotides long, 10-15 nucleotides long, 10-14 nucleotides long, 10-13 nucleotides long, 10-12 nucleotides long, 60 nucleotides long, 59 nucleotides long, 58 nucleotides long, 57 nucleotides long, 56 nucleotides long, 55 nucleotides long, 54 nucleotides long, 53 nucleotides long, 52 nucleotides long, 51 nucleotides long, 50 nucleotides long, 49 nucleotides long, 48 nucleotides long, 47 nucleotides long, 46 nucleotides long, 45 nucleotides long, 44 nucleotides long, 43 nucleotides long, 42 nucleotides long, 41 nucleotides long, 40 nucleotides long, 39 nucleotides long, 38 nucleotides long, 37 nucleotides long, 36 nucleotides long, 35 nucleotides long, 34 nucleotides long, 33 nucleotides long, 32 nucleotides long, 31 nucleotides long, 30 nucleotides long,The lengths may be 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 bases, but are not limited to these lengths, and may be increased or decreased by 1, 2, or 3 bases. 【0052】 The antisense oligomer of the present invention has the activity to simultaneously skip two or more sequentially numbered exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA. In this specification, the skipping of two or more sequentially numbered exons from the target pre-mRNA is referred to as "multi-exon skipping" or "multi-skipping," and this activity is referred to as "multi-exon skipping activity" or "multi-skipping activity." 【0053】 Here, "a sequence of two or more exons selected from the group consisting of exons 45 through 55" refers to multiple exons from the 11 exons from exon 45 through 55 contained in the pre-mRNA, where the exon number increases by 1 each time. The exon number refers to the number assigned to the 79 exons contained in human dystrophin pre-mRNA, with the exon located furthest to the 5' end of the pre-mRNA being designated as the 1st exon, and the exons located sequentially from the 5' end to the 3' end being designated as the 2nd, 3rd, ... 79th exon. In the case of introns, the number is the same as the exon located on their 5' end. That is, the 45th intron has the 45th exon on its 5' end and the 46th exon on its 3' end. Furthermore, in this specification, the "nth" exon or intron refers to the nth exon or intron in the pre-mRNA, counting from the 5' end to the 3' end. 【0054】 Table 4 shows the combinations of exons that are included in any two or more consecutively numbered exons selected from the group consisting of exons 45 through 55. 【0055】 [Table 4] 【0056】 Among the exon combinations listed in Table 4, skipping patterns expected to exhibit higher therapeutic efficacy for DMD include, for example, combinations 1, 2, 3, 4, 6, 8, 10, 18, 20, 21, 23, 25, 27, 28, 30, 32, 34, 36, 38, 40, 41, 43, 45, 46, 50, 52, or 55. By performing multi-exon skipping with these combinations, it is expected that therapeutic effects can be achieved in a larger number of DMD patients. 【0057】 Furthermore, any two or more consecutive exons selected from the group consisting of exons 45 to 55 may include multiple consecutive exon groups, and are not limited to such groups. For example, (Example 1) exons 45-46 (first exon group) and exons 48-53 (second exon group), or (Example 2) exons 46-47 (first exon group), exons 49-50 (second exon group), and exons 52-54 (third exon group). 【0058】 In the present invention, "skipping activity" (i.e., multi-skipping activity) refers to the activity that, taking human dystrophin pre-mRNA as an example, results in human dystrophin mRNA in which two or more consecutive exons selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA are deleted in numerical order. In other words, this activity means that when the antisense oligomer of the present invention binds to a target site on human dystrophin pre-mRNA, and the pre-mRNA undergoes splicing, the nucleotide at the 5' end of the exon immediately downstream of the deleted exon is linked to the nucleotide at the 3' end of the exon immediately upstream of the deleted exon, resulting in the formation of mature mRNA without a codon frameshift (i.e., mature mRNA with an exon deleted without a frameshift). 【0059】 The antisense oligomers of the present invention exhibit multi-skipping activity under physiological conditions. "Physiological conditions" refers to conditions adjusted to similar pH, salt composition, and temperature as those found in living organisms. For example, conditions such as 25-40°C, preferably 37°C, pH 5-8, preferably pH 7.4, and a sodium chloride concentration of 150 mM are used. When inducing multi-skipping, one or more antisense oligomers of the present invention can be used in combination, for example, 2, 3, 4, 5, 6, 7, 8, 9, or 10 types of antisense oligomers. 【0060】 Whether or not multi-skipping has occurred can be confirmed by introducing the antisense oligomer of the present invention into dystrophin-expressing cells (e.g., human rhabdomyosarcoma cells), RT-PCR amplified the region surrounding exons 45-55 of the human dystrophin gene mRNA from the total RNA of the dystrophin-expressing cells, and performing nested PCR or sequencing analysis on the PCR amplification product. Multi-skipping efficiency can be calculated by recovering the human dystrophin gene mRNA from the test cells, measuring the polynucleotide amount "A" of the band in which two or more consecutive exons from exons 45-55 have been skipped, the polynucleotide amount "B" of the band in which one exon from exons 45-55 has been skipped, and the polynucleotide amount "C" of the band in which no skipping has occurred, and then calculating the multi-skipping efficiency according to the following formula based on the measured values ​​of "A", "B", and "C". Skipping efficiency (%) = A / (A + B + C) × 100 【0061】 For example, the multi-skipping efficiency of exons 45-55 can be calculated by measuring the polynucleotide amount "A" of the bands that have undergone multi-skipping of exons 45-55 using a forward primer for exon 44 and a reverse primer for exon 56, measuring the polynucleotide amount "B" of the bands that have undergone single-skipping of exon 45 using a forward primer for exon 44 and a reverse primer for exon 46, and measuring the polynucleotide amount "C" of the bands in which skipping did not occur using a forward primer for exon 44 and a reverse primer for exon 46, and then calculating the efficiency according to the above formula based on these measured values ​​of "A", "B", and "C". 【0062】 The number of exons deleted in human dystrophin mRNA by the antisense oligomer of the present invention is 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11. This is referred to as a deletion pattern, but various deletion patterns may be mixed in the results obtained from a single skipping experiment or skipping treatment. For example, when the antisense oligomer of the present invention is introduced into cells expressing human dystrophin pre-mRNA and the mRNA is recovered, a mixture of mRNA with 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 exons deleted will be obtained. 【0063】 In one embodiment, the "skipping activity" can be defined as (C1) to (C10) below. 【0064】 (C1) Any two consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the two consecutive exons in numerical order may be the 45th and 46th exons, the 46th and 47th exons, the 47th and 48th exons, the 48th and 49th exons, the 49th and 50th exons, the 50th and 51st exons, the 51st and 52nd exons, the 52nd and 53rd exons, the 53rd and 54th exons, or the 54th and 55th exons. 【0065】 (C2) Any three consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the three consecutive exons in numerical order may be the 45th to 47th exons, the 46th to 48th exons, the 47th to 49th exons, the 48th to 50th exons, the 49th to 51st exons, the 50th to 52nd exons, the 51st to 53rd exons, the 52nd to 54th exons, or the 53rd to 55th exons. 【0066】 (C3) Any four consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the four consecutive exons in numerical order may be the 45th to 48th exons, the 46th to 49th exons, the 47th to 50th exons, the 48th to 51st exons, the 49th to 52nd exons, the 50th to 53rd exons, the 51st to 54th exons, or the 52nd to 55th exons. 【0067】 (C4) Any five consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the five consecutive exons in numerical order may be the 45th to 49th exons, the 46th to 50th exons, the 47th to 51st exons, the 48th to 52nd exons, the 49th to 53rd exons, the 50th to 54th exons, or the 51st to 55th exons. 【0068】 (C5) Any six consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the six consecutive exons in numerical order may be the 45th to 50th exons, the 46th to 51st exons, the 47th to 52nd exons, the 48th to 53rd exons, the 49th to 54th exons, or the 50th to 55th exons. 【0069】 (C6) Seven consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the seven consecutive exons in numerical order may be the 45th to 51st exons, the 46th to 52nd exons, the 47th to 53rd exons, the 48th to 54th exons, or the 49th to 55th exons. 【0070】 (C7) Eight consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the eight consecutive exons in numerical order may be the 45th to 52nd exons, the 46th to 53rd exons, the 47th to 54th exons, or the 48th to 55th exons. 【0071】 (C8) Any nine consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the nine consecutive exons in numerical order may be the 45th to 53rd exons, the 46th to 54th exons, or the 47th to 55th exons. 【0072】 (C9) Ten consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA are skipped with efficiencies of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, and 95% or more. Here, the ten consecutive exons in numerical order may be the 45th to 54th exons, or the 46th to 55th exons. 【0073】 Eleven sequentially numbered exons selected from the group consisting of exons 45 through 55 in (C10) human dystrophin pre-mRNA are skipped with efficiencies of 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, and 95% or higher. Here, the 11 consecutive exons in numerical order are the 45th to the 55th exons. 【0074】 The antisense oligomers of the present invention are 10-60 nucleotides long, 10-55 nucleotides long, 10-50 nucleotides long, 10-45 nucleotides long, 10-40 nucleotides long, 10-35 nucleotides long, 10-30 nucleotides long, 10-25 nucleotides long, 15-60 nucleotides long, 15-55 nucleotides long, 15-50 nucleotides long, 15-45 nucleotides long, 15-40 nucleotides long, 15-35 nucleotides long, 15-30 nucleotides long, 15-25 nucleotides long, 16-60 nucleotides long, 16-55 nucleotides long, 16-45 nucleotides long, 16-40 nucleotides long, 16-35 nucleotides long, 16-30 nucleotides long, 16-25 nucleotides long, 17-60 nucleotides long, and 17-55 nucleotides long. , 17-50 nucleotides, 17-45 nucleotides, 17-40 nucleotides, 17-35 nucleotides, 17-30 nucleotides, 17-25 nucleotides, 18-60 nucleotides, 18-55 nucleotides, 18-50 nucleotides, 18-45 nucleotides, 18-40 nucleotides, 18-35 nucleotides, 18-30 nucleotides, 18-25 nucleotides, 19-60 nucleotides, 19-55 nucleotides, 19-50 nucleotides, 19-45 nucleotides, 19-40 nucleotides, 19-35 nucleotides, 19-30 nucleotides, 19-25 nucleotides, 20-60 nucleotides, 20-55 nucleotides, 20-50 nucleotides, 20-45 nucleotides, 20-40 nucleotides, 20-35 Base length, 20-30 base length, 20-25 base length, 15-30 base length, 15-29 base length, 15-28 base length, 15-27 base length, 15-26 base length, 15-25 base length, 15-24 base length, 15-23 base length, 15-22 base length, 15-21 base length, 15-20 base length, 15-19 base length, 15-18 base length, 16-30 base length, 16-29 base length, 16-28 base length, 16-27 base length, 16-26 base length, 16-25 base length, 16-24 base length, 16-23 base length, 16-22 base length, 16-21 base length, 16-20 base length, 16-19 base length, 16 ~18 nucleotides, 17~30 nucleotides, 17~29 nucleotides, 17~28 nucleotides, 17~27 nucleotides, 17~26 nucleotides, 17~25 nucleotides, 17~24 nucleotides, 17~23 nucleotides, 17~22 nucleotides, 17~21 nucleotides, 17~20 nucleotides, 17~19 nucleotides, 17~18 nucleotides, 18~30 nucleotides, 18~29 nucleotides, 18~28 nucleotides, 18~27 nucleotides, 18~26 nucleotides, 18~25 nucleotides, 18~24 nucleotides, 18~23 nucleotides, 18~22 nucleotides, 18~21 nucleotides, 18~20 nucleotides, 18~19 nucleotides, 19~30 nucleotides, 19~29 nucleotides,19-28 nucleotides long, 19-27 nucleotides long, 19-26 nucleotides long, 19-25 nucleotides long, 19-24 nucleotides long, 19-23 nucleotides long, 19-22 nucleotides long, 19-21 nucleotides long, 19-20 nucleotides long, 20-30 nucleotides long, 20-29 nucleotides long, 20-28 nucleotides long, 20-27 nucleotides long, 20-26 nucleotides long, 20-25 nucleotides long, 20-24 nucleotides long, 20-23 nucleotides long, 20-22 nucleotides long, 20-21 nucleotides long, 60 nucleotides long, 59 nucleotides long, 58 nucleotides long, 57 nucleotides long, 56 nucleotides long, 55 nucleotides long, 54 nucleotides long, 53 nucleotides long, 52 nucleotides long, 51 nucleotides long, 50 nucleotides long, 49 nucleotides long, 48 nucleotides long, 47 nucleotides long, 46 nucleotides long, 45 nucleotides long, The lengths may be 44 nucleotides, 43 nucleotides, 42 nucleotides, 41 nucleotides, 40 nucleotides, 39 nucleotides, 38 nucleotides, 37 nucleotides, 36 nucleotides, 35 nucleotides, 34 nucleotides, 33 nucleotides, 32 nucleotides, 31 nucleotides, 30 nucleotides, 29 nucleotides, 28 nucleotides, 27 nucleotides, 26 nucleotides, 25 nucleotides, 24 nucleotides, 23 nucleotides, 22 nucleotides, 21 nucleotides, 20 nucleotides, 19 nucleotides, 18 nucleotides, 17 nucleotides, 16 nucleotides, 15 nucleotides, 14 nucleotides, 13 nucleotides, 12 nucleotides, 11 nucleotides, or 10 nucleotides, but are not limited to these lengths, and may be increased or decreased by 1, 2, or 3 nucleotides. 【0075】 One embodiment of the antisense oligomer of the present invention is a linked antisense oligomer having a structure in which a plurality of unit oligomers are linked together, or a pharmaceutically acceptable salt thereof, or a hydrate thereof (hereinafter referred to as "the linked antisense oligomer of the present invention"). A unit oligomer means each oligomer that constitutes the linked antisense oligomer of the present invention. In other words, it means a portion (unit) that includes a base sequence that undergoes hybridization with a target base sequence having a continuous base sequence when the linked antisense oligomer of the present invention binds to a target base sequence on human dystrophin pre-mRNA. 【0076】 Specifically, the linked antisense oligomer of the present invention is An antisense oligomer in which two or more unit oligomers are linked, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, Each of the unit oligomers described above contains a base sequence complementary to the base sequence of any one region selected from the group consisting of the regions R1 to R24 (regions shown in SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389 which are wild-type, and mutants thereof may also be included), or a base sequence of a part thereof, and the base sequences of the respective unit oligomers are antisense oligomers that are not continuous or overlapping with each other, or pharmaceutically acceptable salts thereof or hydrates thereof. 【0077】 The unit oligomers may be linked via a linker that does not contribute to hybridization, or may be directly linked without a linker. When the unit oligomers are directly linked, the 3'-end of the unit located on the 5'-end side and the 5'-end of the unit located on the 3'-end side form a phosphate bond or any of the following groups. 【Chemical formula】 (In the formula, X represents -OH, -CH2R 1 , -O-CH2R 1 , -S-CH2R 1 , -NR 2 R 3 , or F; R 1 represents H, alkyl; R 2 and R 3 are the same or different and represent H, alkyl, cycloalkyl, or aryl; Y1 represents O, S, CH2, or NR 1 ; Y2 represents O, S, or NR 1 ; Z represents O or S.) 【0078】 In the linked antisense oligomers of the present invention, each unit oligomer may target base sequences contained in different regions within region R1 to R24, or it may target base sequences contained in the same region. Furthermore, in the linked antisense oligomers of the present invention, the base sequences of each unit oligomer are neither continuous nor overlapping. The statement that the base sequences of each unit oligomer are not continuous means that the target base sequences of each unit oligomer constituting the linked antisense oligomer of the present invention are not continuous as base sequences on human dystrophin pre-mRNA. Therefore, when the target base sequences of each unit oligomer are compared with the base sequences of human dystrophin pre-mRNA, at least one base from the human dystrophin pre-mRNA is interposed between the target base sequences. On the other hand, the statement that the base sequences of each unit oligomer are not overlapping means that the target base sequences of the unit oligomers are not overlapping as base sequences on human dystrophin pre-mRNA. It is acceptable for there to be accidental similarities of approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bases in the base sequence of each unit oligomer. 【0079】 The linked antisense oligomer of the present invention is such that each of the unit oligomers is (D1) One base sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389. (D2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±15% of the length of the selected nucleotide sequence. (D3) A nucleotide sequence that hybridizes under stringent conditions with a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 233-256, 341-369, and 385-389, or (D4) A partial base sequence of any one base sequence selected from the group consisting of (D1), (D2), and (D3) above. It may have a structure that includes a base sequence complementary to the above, and the base sequences of each unit oligomer are not continuous or overlapping with each other. 【0080】 As described above, the linked antisense oligomer of the present invention may include a base sequence complementary to any one base sequence selected from the group consisting of (D1), (D2), (D3), and (D4). The base sequence of (D2) is a variant of the base sequence of (D1), and examples thereof include (D2-1) A base sequence having 85% or more sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±15% compared to the length of any one base sequence selected. (D2-2) A base sequence having 86% or more sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±14% compared to the length of any one base sequence selected. (D2-3) A base sequence having 87% or more sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±13% compared to the length of any one base sequence selected. (D2-4) A base sequence having 88% or more sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±12% compared to the length of any one base sequence selected. (D2-5) A base sequence having 89% or more sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±11% compared to the length of any one base sequence selected. (D2-6) A base sequence having at least 90% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±10% compared to the length of any one selected base sequence. (D2-7) A base sequence having at least 91% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±9% compared to the length of any one selected base sequence. (D2-8) A base sequence having at least 92% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±8% compared to the length of any one selected base sequence. (D2-9) A base sequence having at least 93% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±7% compared to the length of any one selected base sequence. (D2-10) A base sequence having at least 94% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±6% compared to the length of any one selected base sequence. (D2-11) A base sequence having at least 95% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±5% compared to the length of any one selected base sequence. (D2-12) A base sequence having at least 96% sequence identity with any one base sequence selected from the group consisting of SEQ ID NOs: 233 to 256, 341 to 369, and 385 to 389, and having a length within ±4% compared to the length of any one selected base sequence. (D2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±3% of the length of the selected nucleotide sequence. (D2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±2% of the length of the selected nucleotide sequence. (D2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±1% of the length of the selected nucleotide sequence, and (D2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 233-256, 341-369, and 385-389, and having a length within ±0.5% of the length of the selected nucleotide sequence, This can also be mentioned. 【0081】 Therefore, each of the unit oligomers constituting the linked antisense oligomer of the present invention may contain a base sequence complementary to any one base sequence selected from the group consisting of (D1), (D2-1) to (D2-16), (D3), and (D4). 【0082】 Furthermore, as described above, the linked antisense oligomer of the present invention may contain a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of (D1), (D2), (D3), and (D4). The nucleotide sequence (D4) relates to a nucleotide sequence of a portion of any one nucleotide sequence selected from the group consisting of (D1), (D2), and (D3). With respect to (D4), "a portion thereof" means a continuous subregion excluding the entire length of (D1), (D2), or (D3). The length of the subregion is 5 to 25 nucleotides, and also 5 to 24 nucleotides, 5 to 23 nucleotides, 5 to 22 nucleotides, 5 to 21 nucleotides, 5 to 20 nucleotides, 5 to 19 nucleotides, 5 to 18 nucleotides, 5 to 17 nucleotides, 5 to 16 nucleotides, 5 to 15 nucleotides, 5 to 14 nucleotides, 5 to 13 nucleotides, 5 to 12 nucleotides, 7 to 25 nucleotides, 7 to 24 nucleotides, 7 to 23 nucleotides, 7 to 22 nucleotides, and 7 to 21 nucleotides. Length, 7-20 nucleotides, 7-19 nucleotides, 7-18 nucleotides, 7-17 nucleotides, 7-16 nucleotides, 7-15 nucleotides, 7-14 nucleotides, 7-13 nucleotides, 7-12 nucleotides, 9-25 nucleotides, 9-24 nucleotides, 9-23 nucleotides, 9-22 nucleotides, 9-21 nucleotides, 9-20 nucleotides, 9-19 nucleotides, 9-18 nucleotides, 9-17 nucleotides, 9-16 nucleotides, 9-15 nucleotides, 9 ~14 nucleotides, 9~13 nucleotides, 9~12 nucleotides, 10~25 nucleotides, 10~24 nucleotides, 10~23 nucleotides, 10~22 nucleotides, 10~21 nucleotides, 10~20 nucleotides, 10~19 nucleotides, 10~18 nucleotides, 10~17 nucleotides, 10~16 nucleotides, 10~15 nucleotides, 10~14 nucleotides, 10~13 nucleotides, 10~12 nucleotides, 25 nucleotides, 24 nucleotides, 23 The length may be 1 / 2, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 bases, but is not limited to these lengths, and may be increased or decreased by 1, 2, or 3 bases. 【0083】 The size of each unit oligomer ranges from 5 to 30 nucleotides in length, including 5-29 nucleotides, 5-28 nucleotides, 5-27 nucleotides, 5-26 nucleotides, 5-25 nucleotides, 5-24 nucleotides, 5-23 nucleotides, 5-22 nucleotides, 5-21 nucleotides, 5-20 nucleotides, 5-19 nucleotides, 5-18 nucleotides, 5-17 nucleotides, 5-16 nucleotides, 5-15 nucleotides, 5-14 nucleotides, 5-13 nucleotides, 5-12 nucleotides, 7-30 nucleotides, 7-29 nucleotides, 7-28 nucleotides, 7-27 nucleotides, 7-26 nucleotides, and 7- 25 nucleotides, 7-24 nucleotides, 7-23 nucleotides, 7-22 nucleotides, 7-21 nucleotides, 7-20 nucleotides, 7-19 nucleotides, 7-18 nucleotides, 7-17 nucleotides, 7-16 nucleotides, 7-15 nucleotides, 7-14 nucleotides, 7-13 nucleotides, 7-12 nucleotides, 9-30 nucleotides, 9-29 nucleotides, 9-28 nucleotides, 9-27 nucleotides, 9-26 nucleotides, 9-25 nucleotides, 9-24 nucleotides, 9-23 nucleotides, 9-22 nucleotides, 9-21 nucleotides, 9-20 nucleotides, 9-19 nucleotides, 9-18 nucleotides , 9-17 nucleotide lengths, 9-16 nucleotide lengths, 9-15 nucleotide lengths, 9-14 nucleotide lengths, 9-13 nucleotide lengths, 9-12 nucleotide lengths, 10-30 nucleotide lengths, 10-29 nucleotide lengths, 10-28 nucleotide lengths, 10-27 nucleotide lengths, 10-26 nucleotide lengths, 10-25 nucleotide lengths, 10-24 nucleotide lengths, 10-23 nucleotide lengths, 10-22 nucleotide lengths, 10-21 nucleotide lengths, 10-20 nucleotide lengths, 10-19 nucleotide lengths, 10-18 nucleotide lengths, 10-17 nucleotide lengths, 10-16 nucleotide lengths, 10-15 nucleotide lengths, 10-14 nucleotide lengths, 10-13 nucleotide lengths, 10 The lengths may be ~12 nucleotides, 30 nucleotides, 29 nucleotides, 28 nucleotides, 27 nucleotides, 26 nucleotides, 25 nucleotides, 24 nucleotides, 23 nucleotides, 22 nucleotides, 21 nucleotides, 20 nucleotides, 19 nucleotides, 18 nucleotides, 17 nucleotides, 16 nucleotides, 15 nucleotides, 14 nucleotides, 13 nucleotides, 12 nucleotides, 11 nucleotides, 10 nucleotides, 9 nucleotides, 8 nucleotides, 7 nucleotides, 6 nucleotides, or 5 nucleotides, but are not limited to these lengths, and may be increased or decreased by 1, 2, or 3 nucleotides. The size of each unit oligomer may be the same or different. 【0084】 In one embodiment, the linked antisense oligomer of the present invention is (E1) Any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, or (E2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±15% of the length of the selected nucleotide sequence. This may be an antisense oligomer consisting of the above, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0085】 Here, the base sequence of (E2) is a variant of the base sequence of (E1), but as a variant, (E2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±15% of the length of the selected nucleotide sequence. (E2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±14% of the length of the selected nucleotide sequence. (E2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±13% of the length of the selected nucleotide sequence. (E2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±12% of the length of the selected nucleotide sequence. (E2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±11% of the length of the selected nucleotide sequence. (E2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±10% of the length of the selected nucleotide sequence. (E2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±9% of the length of the selected nucleotide sequence. (E2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±8% of the length of the selected nucleotide sequence. (E2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±7% of the length of the selected nucleotide sequence. (E2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±6% of the length of the selected nucleotide sequence. (E2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±5% of the length of the selected nucleotide sequence. (E2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±4% of the length of the selected nucleotide sequence. (E2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±3% of the length of the selected nucleotide sequence. (E2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±2% of the length of the selected nucleotide sequence. (E2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and having a length within ±1% of the length of the selected nucleotide sequence, (E2-16) A nucleotide sequence that has 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 18-21, 80-111, and 225-232, and whose length is within ±0.5% of the length of the selected nucleotide sequence. 【0086】 Therefore, the linked antisense oligomer of the present invention may consist of any one base sequence selected from the group consisting of (E1) and (E2-1) to (E2-16). 【0087】 Preferably, the linked antisense oligomer of the present invention contains any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 18-21, 80-111, and 225-232. In some embodiments, the linked antisense oligomer of the present invention consists of any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 18-21, 80-111, and 225-232. 【0088】 Furthermore, the antisense oligomer of the present invention is (E'1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, or (E'2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±15% of the length of the selected nucleotide sequence. This may be an antisense oligomer consisting of the above, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0089】 Here, the base sequence of (E'2) is a variant of the base sequence of (E'1), but as a variant, (E'2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±15% of the length of the selected nucleotide sequence. (E'2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±14% of the length of the selected nucleotide sequence. (E'2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±13% of the length of the selected nucleotide sequence. (E’2-4) A base sequence having at least 88% sequence identity with any one base sequence selected from the group consisting of SEQ ID NO: 1 to 111, 113 to 177, 179, 180, 182, 183, 185 to 193, 195 to 198, 200 to 223, and 225 to 232, and having a length within ±12% compared to the length of any one base sequence selected, (E’2-5) A base sequence having at least 89% sequence identity with any one base sequence selected from the group consisting of SEQ ID NO: 1 to 111, 113 to 177, 179, 180, 182, 183, 185 to 193, 195 to 198, 200 to 223, and 225 to 232, and having a length within ±11% compared to the length of any one base sequence selected, (E’2-6) A base sequence having at least 90% sequence identity with any one base sequence selected from the group consisting of SEQ ID NO: 1 to 111, 113 to 177, 179, 180, 182, 183, 185 to 193, 195 to 198, 200 to 223, and 225 to 232, and having a length within ±10% compared to the length of any one base sequence selected, (E’2-7) A base sequence having at least 91% sequence identity with any one base sequence selected from the group consisting of SEQ ID NO: 1 to 111, 113 to 177, 179, 180, 182, 183, 185 to 193, 195 to 198, 200 to 223, and 225 to 232, and having a length within ±9% compared to the length of any one base sequence selected, (E’2-8) A base sequence having at least 92% sequence identity with any one base sequence selected from the group consisting of SEQ ID NO: 1 to 111, 113 to 177, 179, 180, 182, 183, 185 to 193, 195 to 198, 200 to 223, and 225 to 232, and having a length within ±8% compared to the length of any one base sequence selected, (E'2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±7% of the length of the selected nucleotide sequence. (E'2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±6% of the length of the selected nucleotide sequence. (E'2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±5% of the length of the selected nucleotide sequence. (E'2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±4% of the length of the selected nucleotide sequence. (E'2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±3% of the length of the selected nucleotide sequence. (E'2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±2% of the length of the selected nucleotide sequence. (E'2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±1% of the length of the selected nucleotide sequence, and (E'2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±0.5% of the length of the selected nucleotide sequence, This can also be mentioned. 【0090】 Therefore, the antisense oligomer of the present invention may consist of any one base sequence selected from the group consisting of (E'1) and (E'2-1) to (E'2-16). 【0091】 Preferably, the antisense oligomer of the present invention contains any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232. In some embodiments, the antisense oligomer of the present invention consists of any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232. 【0092】 In one embodiment, the antisense oligomer of the present invention is (E''1) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 11 Any one base sequence selected from the group consisting of 9, 121, 122, 124, 125, 126, 128, 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, or (E''2) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 13 A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of 0, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±15% of the length of the selected nucleotide sequence. This may be an antisense oligomer consisting of the above, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0093】 Here, the base sequence of (E''2) is a variant of the base sequence of (E''1), but as a variant, (E''2-1) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±15% of the length of the selected nucleotide sequence. (E''2-2) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±14% of the length of the selected nucleotide sequence. (E''2-3) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±13% of the length of the selected nucleotide sequence. (E''2-4) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±12% of the length of the selected nucleotide sequence. (E''2-5) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±11% of the length of the selected nucleotide sequence. (E''2-6) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±10% of the length of the selected nucleotide sequence. (E''2-7) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±9% of the length of the selected nucleotide sequence. (E''2-8) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±8% of the length of the selected nucleotide sequence. (E''2-9) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±7% of the length of the selected nucleotide sequence. (E''2-10) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±6% of the length of the selected nucleotide sequence. (E''2-11) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±5% of the length of the selected nucleotide sequence. (E''2-12) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±4% of the length of the selected nucleotide sequence. (E''2-13) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±3% of the length of the selected nucleotide sequence. (E''2-14) Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±2% of the length of the selected nucleotide sequence. (E''2-15) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 1 A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of 30, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±1% of the length of the selected nucleotide sequence, and (E''2-16) Sequence IDs 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 13 A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of 0, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0094】 Therefore, the antisense oligomer of the present invention may consist of any one base sequence selected from the group consisting of (E''1) and (E''2-1) to (E''2-16). 【0095】 In one embodiment, the antisense oligomer of the present invention is (E'''1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, or (E'''2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±15% of the length of the selected nucleotide sequence. This may be an antisense oligomer consisting of the above, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0096】 Here, the base sequence (E'''2) is a variant of the base sequence (E'''1), but as a variant, (E'''2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±15% of the length of the selected nucleotide sequence. (E'''2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±14% of the length of the selected nucleotide sequence. (E'''2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±13% of the length of the selected nucleotide sequence. (E'''2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of Sequence IDs 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±12% of the length of the selected nucleotide sequence. (E'''2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±11% of the length of the selected nucleotide sequence. (E'''2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±10% of the length of the selected nucleotide sequence. (E'''2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±9% of the length of the selected nucleotide sequence. (E'''2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±8% of the length of the selected nucleotide sequence. (E'''2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±7% of the length of the selected nucleotide sequence. (E'''2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±6% of the length of the selected nucleotide sequence. (E'''2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±5% of the length of the selected nucleotide sequence. (E'''2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±4% of the length of the selected nucleotide sequence. (E'''2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±3% of the length of the selected nucleotide sequence. (E'''2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±2% of the length of the selected nucleotide sequence. (E'''2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±1% of the length of the selected nucleotide sequence, and (E'''2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0097】 Therefore, the antisense oligomer of the present invention may consist of any one base sequence selected from the group consisting of (E'''1) and (E'''2-1) to (E'''2-16). 【0098】 Preferably, the antisense oligomer of the present invention is SEQ ID NOs: 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 1 An antisense oligomer containing any one base sequence selected from the group consisting of 24, 125, 126, 128, 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. Furthermore, in a certain embodiment, the antisense oligomer of the present invention is SEQ ID NOs: 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 1 An antisense oligomer consisting of any one base sequence selected from the group consisting of 24, 125, 126, 128, 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0099】 More preferably, the antisense oligomer of the present invention is an antisense oligomer containing any one base sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. In some embodiments, the antisense oligomer of the present invention is an antisense oligomer consisting of any one base sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0100】 In some embodiments, the linked antisense oligomer of the present invention is an antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 80, 82, 86, 92, 97, 98, 100, 102, 225, 228, 231, and 232. In some embodiments, the linked antisense oligomer of the present invention comprises any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 80, 82, 86, 92, 97, 98, 100, 102, 225, 228, 231, and 232. More preferably, the linked antisense oligomer of the present invention comprises any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 102, 225, and 228. In one embodiment, the linked antisense oligomer of the present invention is an antisense oligomer consisting of any one base sequence selected from the group consisting of SEQ ID NOs: 102, 225, and 228, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0101】 The antisense oligomer of the present invention contains a nucleotide sequence complementary to the nucleotide sequence of at least one region selected from the group consisting of regions R1 to R24, or a nucleotide sequence of a part thereof. As the nucleotide sequence complementary to the nucleotide sequence of each region included in regions R1 to R24, any nucleotide sequence selected from the group consisting of (E1) and (E2-1) to (E2-16), the group consisting of (E'1) and (E'2-1) to (E'2-16), the group consisting of (E''1) and (E''2-1) to (E''2-16), or the group consisting of (E'''1) and (E'''2-1) to (E'''2-16) may be selected. For example, the base sequences for regions R1 to R24, or parts thereof, may be complementary base sequences to the base sequences of each region or parts thereof, and for regions R1 to R3, they may be base sequences selected from the group consisting of (E1) and (E2-1) to (E2-16), which are included in the regions R1 to R3. 【0102】 The antisense oligomer of the present invention (including the linked antisense oligomer of the present invention) may be an oligonucleotide, a morpholino oligomer, or a peptide nucleic acid (PNA) oligomer (hereinafter, these will also be referred to as "the antisense oligonucleotide of the present invention," "the antisense morpholino oligomer of the present invention," or "the antisense peptide nucleic acid oligomer of the present invention," respectively). 【0103】 The antisense oligonucleotide of the present invention is an antisense oligomer having a nucleotide as a constituent unit, and such nucleotide may be a ribonucleotide, a deoxyribonucleotide, or a modified nucleotide. 【0104】 A modified nucleotide refers to a ribonucleotide or deoxyribonucleotide in which all or part of the nucleic acid base, sugar moiety, and phosphate bond moiety are modified. 【0105】 Examples of nucleic acid bases include adenine, guanine, hypoxanthine, cytosine, thymine, uracil, or modified bases thereof. Examples of such modified bases include pseudouracil, 3-methyluracil, dihydrouracil, 5-alkylcytosine (e.g., 5-methylcytosine), 5-alkyluracil (e.g., 5-ethyluracil), 5-halouracil (e.g., 5-bromouracil), 6-azapyrimidine, 6-alkylpyrimidine (e.g., 6-methyluracil), 2-thiouracil, 4-thiouracil, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, 1-methyladenine, 1-methylhypoxanthine Examples include, but are not limited to, santin, 2,2-dimethylguanine, 3-methylcytosine, 2-methyladenine, 2-methylguanine, N6-methyladenine, 7-methylguanine, 5-methoxyaminomethyl-2-thiouracil, 5-methylaminomethyluracil, 5-methylcarbonylmethyluracil, 5-methyloxyuracil, 5-methyl-2-thiouracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid, 2-thiocytosine, purines, 2,6-diaminopurines, 2-aminopurines, isoguanine, indole, imidazole, xanthine, etc. 【0106】 Modifications of the sugar portion include, for example, modifications of the 2' position of ribose and modifications of other parts of the sugar. Modifications of the 2' position of ribose include, for example, the substitution of the -OH group at the 2' position of ribose with -OR, -R, -R'OR, -SH, -SR, -NH2, -NHR, -NR2, -N3, -CN, -F, -Cl, -Br, or -I. Here, R represents alkyl or aryl, and R' represents alkylene. Other modifications of the sugar include, but are not limited to, substitution of the 4'-position O of ribose or deoxyribose with S, and bridging of the 2' and 4' positions of the sugar, such as LNA (Locked Nucleic Acid) or ENA (2'-O,4'-C-Ethylene-bridged Nucleic Acids). 【0107】 Modifications of the phosphate bond include, for example, the substitution of the phosphodiester bond with a phosphorothioate bond, phosphorodithioate bond, alkylphosphonate bond, phosphoramidate bond, or boranophosphate bond (see, for example, Enya et al: Bioorganic & Medicinal Chemistry, 2008, 18, 9154-9160) (see, for example, republished patent applications 2006 / 129594 and 2006 / 038608). 【0108】 In this specification, linear or branched alkyl groups having 1 to 6 carbon atoms are preferred. Specifically, examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, and isohexyl. The alkyl group may be substituted, and examples of such substituents include halogens, alkoxys, cyanos, and nitros, and may be substituted with 1 to 3 of these substituents. 【0109】 In this specification, cycloalkyls having 3 to 12 carbon atoms are preferred. Specifically, examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, and cyclododecyl. 【0110】 In this specification, examples of halogens include fluorine, chlorine, bromine, and iodine. 【0111】 In this specification, examples of alkoxys include linear or branched alkoxys having 1 to 6 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentyloxy, isopentyloxy, n-hexyloxy, and isohexyloxy. In particular, alkoxys having 1 to 3 carbon atoms are preferred. 【0112】 In this specification, aryls having 6 to 10 carbon atoms are preferred. Specifically, examples include phenyl, α-naphthyl, and β-naphthyl. Phenyne is particularly preferred. The aryl may be substituted, and examples of such substituents include alkyl, halogen, alkoxy, cyano, and nitro atoms, and one to three of these may be substituted. 【0113】 In this specification, linear or branched alkylenes having 1 to 6 carbon atoms are preferred. Specifically, examples include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, 2-(ethyl)trimethylene, and 1-(methyl)tetramethylene. 【0114】 In this specification, acyls can be linear or branched alkanoyls or aroyls. Examples of alkanoyls include formyl, acetyl, 2-methylacetyl, 2,2-dimethylacetyl, propionyl, butyryl, isobutyryl, pentanoyl, 2,2-dimethylpropionyl, and hexanoyl. Examples of aroyls include benzoyl, toluyl, and naphthoyl. Such aroyls may be substituted at substitutable positions, or they may be substituted with alkyl groups. 【0115】 The antisense oligonucleotide of the present invention is preferably an antisense oligomer of the present invention having a group represented by the following general formula as a constituent unit, wherein the -OH group at the 2' position of ribose is substituted with methoxy, and the phosphate bond portion is a phosphorothioate bond. [ka] (In the formula, Base represents a nucleic acid base.) 【0116】 The antisense oligonucleotides of the present invention can be easily synthesized using various automated synthesis equipment (e.g., AKTA oligopilot plus 10 / 100 (GE Healthcare)), or they can be manufactured by a third-party organization (e.g., Promega or Takara). 【0117】 The antisense morpholino oligomer of the present invention is an antisense oligomer having a group represented by the following general formula as its constituent unit. [ka] (In the formula, Base is synonymous with the above; W represents a group that can be expressed by one of the following formulas. [ka] (In the formula, X is -CH2R 1 , -O-CH2R 1 -S-CH2R 1 , -NR 2 R 3 , or represents F; R 1 H represents alkyl; R 2 and R 3 These represent H, alkyl, cycloalkyl, or aryl, either identical or distinct; Y1 is O, S, CH2, or NR 1 It represents; Y2 is O, S, or NR 1 It represents; Z represents either O or S. 【0118】 Examples of morpholino monomer compounds used to synthesize the antisense morpholino oligomer of the present invention include, but are not limited to, the morpholino monomer compounds (A), (C), (T), and (G) shown in Table 5. 【0119】 [Table 5] 【0120】 In the present invention, the morpholino oligomer is preferably an oligomer (phosphodiamide morpholino oligomer (hereinafter referred to as "PMO")) whose constituent unit is a group represented by the following formula. [ka] (In the formula, Base, R 2 , R 3 (This is synonymous with the above.) 【0121】 Morpholino oligomers can be manufactured, for example, according to the methods described in International Publication No. 1991 / 009033 or International Publication No. 2009 / 064471. In particular, PMOs can be manufactured according to the methods described in International Publication No. 2009 / 064471 or International Publication No. 2013 / 100190. 【0122】 The antisense peptide nucleic acid oligomer of the present invention is an antisense oligomer having a group represented by the following general formula as its constituent unit. [ka] (In the formula, Base has the same meaning as above.) Peptide nucleic acid oligomers can be manufactured, for example, according to the following literature. 1)PE Nielsen, M. Egholm, RH Berg, O. Buchardt,Science, 254, 1497 (1991)2)M. Egholm, O. Buchardt, PE Nielsen, RH Berg,JACS, 114, 1895 (1992)3)KL Dueholm, M. Egholm, C. Behrens, L. Christensen, HF Hansen, T. Vulpius, KH Petersen, RH Berg, PE Nielsen, O. Buchardt,J. Org. Chem., 59, 5767 (1994) 4)L. Christensen, R. Fitzpatrick, B. Gildea, KH Petersen, HF Hansen, T. Koch, M. Egholm,O. Buchardt, PE Nielsen, J. Coull, RH Berg, J. Pept. Sci., 1, 175 (1995)5)T. Koch, HF Hansen, P. Andersen, T. Larsen, HG Batz, K. Otteson, H. Orum, J. Pept. Res., 49, 80 (1997) 【0123】 The antisense oligomer of the present invention (including the linked antisense oligomer of the present invention) may be in the form of a pharmaceutically acceptable salt, a hydrate, or a pharmaceutically acceptable salt hydrate. 【0124】 Examples of pharmaceutically acceptable salts of the antisense oligomer of the present invention include alkali metal salts such as sodium salts, potassium salts, and lithium salts; alkaline earth metal salts such as calcium salts and magnesium salts; metal salts such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, and cobalt salts; ammonium salts; t-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, N-benzylphenethylamine salts, and pipette salts. Examples include organic amine salts such as radin salts, tetramethylammonium salts, and tris(hydroxymethyl)aminomethane salt; hydrohalides such as hydrofluoric acid, hydrochloride, hydrobromide, and hydroiodide; inorganic salts such as nitrates, perchlorates, sulfates, and phosphates; lower alkanesulfonates such as methanesulfonates, trifluoromethanesulfonates, and ethanesulfonates; arylsulfonates such as benzenesulfonates and p-toluenesulfonates; organic salts such as acetates, malates, fumarates, succinates, citrates, tartrates, oxalates, and maleates; and amino acid salts such as glycine salts, lysine salts, arginine salts, ornithine salts, glutamates, and aspartates. These salts can be produced by known methods. Alternatively, the antisense oligomers of the present invention may also be in the form of hydrates. 【0125】 2. Suppressor antisense oligomer In another embodiment of the present invention, a suppressor antisense oligomer is provided that suppresses single-exon skipping (hereinafter referred to as "single skipping"). By suppressing single skipping, the suppressor antisense oligomer can enhance the multi-exon skipping effect of the antisense oligomer. Single skipping refers to the skipping of only one exon, rather than skipping multiple exons simultaneously. 【0126】 Specifically, the present invention provides a suppressor antisense oligomer, a pharmaceutically acceptable salt thereof, or a hydrate thereof that inhibits single skipping of any one exon selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA. Hereinafter, the suppressor antisense oligomer, its pharmaceutically acceptable salt, or its hydrate shall be collectively referred to as "the suppressor antisense oligomer of the present invention." The suppressor antisense oligomer of the present invention may also refer to the suppressor antisense oligomer, its pharmaceutically acceptable salt, or its hydrate individually. 【0127】 The suppressor antisense oligomer of the present invention inhibits single skipping by targeting splicing silencer sequences, branch site sequences, or splice site sequences on human dystrophin pre-mRNA and inhibiting splicing. The suppressor antisense oligomer of the present invention reduces the efficiency of single skipping of the target exon compared to the control. 【0128】 For example, a suppressor antisense oligomer that inhibits single skipping of any one exon selected from the group consisting of exons 45 to 55 targets a splicing silencer sequence of any exon between 44 and 56, or a splicing silencer sequence, branch site sequence, or splice site sequence of any intron between 44 and 55. 【0129】 In this specification, a splicing silencer sequence is a nucleotide sequence element on pre-mRNA that has the function of suppressing exon recognition. Examples of splicing silencer sequences include recognition sequences for proteins or protein complexes such as heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), hnRNP A2 / B1, DAZAP1, hnRNP I, Fox-1, Fox-2, hnRNP H1, hnRNP H2, hnRNP H3, hnRNP L, Sam68, and SRp40. 【0130】 In this specification, the splice site is the boundary between an exon and an intron. The splice site can be estimated from the fact that the 5' end of an intron begins with GU, AU, etc., and the 3' end of an intron ends with AG, AC, etc. 【0131】 In this specification, a branch site is a sequence within an intron that attacks and covalently bonds to the 5' splice site during a splicing reaction. A branch site sequence refers to the base sequence within an intron that contains the branch site. 【0132】 Examples of splicing silencer sequences, branch site sequences, or splice site sequences that serve as targets (hereinafter referred to as "suppressor sequences") for the suppressor antisense oligomer of the present invention include the following nucleotide sequences (SEQ ID NOs: 370-384 and 390). Furthermore, the locations of the suppressor sequence, branch site sequence, and splice site sequence are as follows: SpliceAid (Francesco Piva et al., Bioinformatics, 25(9), 1211-1213, 2009), SpliceAid-2 (Francesco Piva et al., Human Mutation, 33(1), 81-85, 2012), SpliceAid-F (Matteo Giulietti et al., Nucleic Acids Res., 41, D125-D131, 2012), ATtRACT-a (Girolamo Giudice et al., Database (Oxford), baw035, 2016), SROOGLE (Schraga Schwartz et al., Nucleic Acids Res., 37, W189-W192, 2009), Reg RNA (Hsi-Yuan It can be detected using software such as Huang et al., Nucleic Acids Res., 34, W429-W434, 2006), Reg RNA 2.0 (Tzu-Hao Chang et al., BMC bioinformatics, 14(Suppl 2), S4, 2013), Human Splicing Finder (Francois-Olivier Desmet et al., Nucleic Acids Res., 37, 9, e67, 2009), and SVM-BPfinder (Andre Corvelo et al., PLoS Comput Biol., 6, 11, e1001016, 2010). 【0133】 [Table 6] 【0134】 Here, specific examples of splicing silencer sequences include sequence numbers 370-382. Furthermore, as variants of sequence numbers 370-382, (F1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±15% of the length of the selected nucleotide sequence. (F2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±14% of the length of the selected nucleotide sequence. (F3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±13% of the length of the selected nucleotide sequence. (F4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±12% of the length of the selected nucleotide sequence. (F5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±11% of the length of the selected nucleotide sequence. (F6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±10% of the length of the selected nucleotide sequence. (F7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±9% of the length of the selected nucleotide sequence. (F8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±8% of the length of the selected nucleotide sequence. (F9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±7% of the length of the selected nucleotide sequence. (F10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±6% of the length of the selected nucleotide sequence. (F11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±5% of the length of the selected nucleotide sequence. (F12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±4% of the length of the selected nucleotide sequence. (F13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±3% of the length of the selected nucleotide sequence. (F14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±2% of the length of the selected nucleotide sequence. (F15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±1% of the length of the selected nucleotide sequence, (F16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 382, ​​and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0135】 In one aspect of the present invention, the splicing silencer sequence may be a recognition sequence for heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1). Examples of human hnRNP A1 recognition sequences are shown in SEQ ID NOs. 370-379. 【0136】 Furthermore, specific examples of splice site sequences include sequence numbers 384 or 390. Also, as variants of sequence numbers 384 or 390, (G1) A nucleotide sequence having 80% or more sequence identity with the nucleotide sequence of sequence number 384 or 390, and having a length within ±20% of the length of any one of the selected nucleotide sequences, is also an example. 【0137】 Furthermore, as a specific example of a branched region sequence, sequence number 383 can be cited. Also, as a mutant of sequence number 383, (H1) A nucleotide sequence having 80% or more sequence identity with the nucleotide sequence of sequence number 383, and having a length within ±20% of the length of any one of the selected nucleotide sequences. This can also be mentioned. 【0138】 Furthermore, the following target regions are also included in the target base sequence of the suppressor antisense oligomer of the present invention as variants of the above suppressor sequence. (I1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±15% of the length of the selected nucleotide sequence. (I2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±14% of the length of the selected nucleotide sequence. (I3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±13% of the length of the selected nucleotide sequence. (I4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±12% of the length of the selected nucleotide sequence. (I5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±11% of the length of the selected nucleotide sequence. (I6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±10% of the length of the selected nucleotide sequence. (I7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±9% of the length of the selected nucleotide sequence. (I8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±8% of the length of the selected nucleotide sequence. (I9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±7% of the length of the selected nucleotide sequence. (I10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±6% of the length of the selected nucleotide sequence. (I11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±5% of the length of the selected nucleotide sequence. (I12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±4% of the length of the selected nucleotide sequence. (I13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±3% of the length of the selected nucleotide sequence. (I14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±2% of the length of the selected nucleotide sequence. (I15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±1% of the length of the selected nucleotide sequence, (I16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370-384 and 390, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0139】 One embodiment of the suppressor antisense oligomer of the present invention includes a nucleotide sequence complementary to one of the nucleotide sequences selected from the group consisting of the nucleotide sequences shown in SEQ ID NOs. 370-384 and 390 and the mutant nucleotide sequences shown in (I1)-(I16) above. Another embodiment of the present invention includes a nucleotide sequence complementary to one of the nucleotide sequences selected from the group consisting of the nucleotide sequences shown in SEQ ID NOs. 370-384 and 390, or a nucleotide sequence complementary to a nucleotide sequence. Here, "a part thereof" is defined the same as in the section on antisense oligomers in this invention. 【0140】 Further embodiments of the suppressor antisense oligomer of the present invention include: (J1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 257-275, or (J2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±15% of the length of the selected nucleotide sequence. Examples include suppressor antisense oligomers comprising the above, or pharmaceutically acceptable salts thereof, or hydrates thereof. 【0141】 Here, the base sequence of (J2) is a variant of the base sequence of (J1), but as a variant, (J2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±15% of the length of the selected nucleotide sequence. (J2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±14% of the length of the selected nucleotide sequence. (J2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±13% of the length of the selected nucleotide sequence. (J2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±12% of the length of the selected nucleotide sequence. (J2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±11% of the length of the selected nucleotide sequence. (J2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±10% of the length of the selected nucleotide sequence. (J2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±9% of the length of the selected nucleotide sequence. (J2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±8% of the length of the selected nucleotide sequence. (J2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±7% of the length of the selected nucleotide sequence. (J2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±6% of the length of the selected nucleotide sequence. (J2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±5% of the length of the selected nucleotide sequence. (J2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±4% of the length of the selected nucleotide sequence. (J2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±3% of the length of the selected nucleotide sequence. (J2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±2% of the length of the selected nucleotide sequence. (J2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±1% of the length of the selected nucleotide sequence, and (J2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 257 to 275, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0142】 Therefore, further embodiments of the suppressor antisense oligomer of the present invention include a suppressor antisense oligomer comprising any one base sequence selected from the group consisting of (J1) and (J2-1) to (J2-16), or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0143】 Further embodiments of the suppressor antisense oligomer of the present invention include: (J'1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 260, 261, and 263, or (J'2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±15% of the length of the selected nucleotide sequence. Examples include suppressor antisense oligomers comprising the above, or pharmaceutically acceptable salts thereof, or hydrates thereof. 【0144】 Here, the base sequence of (J'2) is a variant of the base sequence of (J'1), but as a variant, (J'2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±15% of the length of the selected nucleotide sequence. (J'2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±14% of the length of the selected nucleotide sequence. (J'2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±13% of the length of the selected nucleotide sequence. (J'2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±12% of the length of the selected nucleotide sequence. (J'2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±11% of the length of the selected nucleotide sequence. (J'2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±10% of the length of the selected nucleotide sequence. (J'2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±9% of the length of the selected nucleotide sequence. (J'2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±8% of the length of the selected nucleotide sequence. (J'2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±7% of the length of the selected nucleotide sequence. (J'2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±6% of the length of the selected nucleotide sequence. (J'2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±5% of the length of the selected nucleotide sequence. (J'2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±4% of the length of the selected nucleotide sequence. (J'2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±3% of the length of the selected nucleotide sequence. (J'2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±2% of the length of the selected nucleotide sequence. (J'2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±1% of the length of the selected nucleotide sequence, (J'2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 260, 261, and 263, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0145】 Therefore, further embodiments of the suppressor antisense oligomer of the present invention include a suppressor antisense oligomer comprising any one base sequence selected from the group consisting of (J'1) and (J'2-1) to (J'2-16), or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0146】 The suppressor antisense oligomer of the present invention enhances the multi-skipping effect under physiological conditions. "Physiological conditions" refers to conditions adjusted to similar pH, salt composition, and temperature as those found in living organisms. For example, conditions such as 25-40°C, preferably 37°C, pH 5-8, preferably pH 7.4, and a sodium chloride concentration of 150 mM are used. 【0147】 Whether the suppressor antisense oligomer of the present invention enhances the multi-exon skipping effect can be confirmed by preparing two experimental systems under the same conditions: (i) a multi-exon skipping experimental system using only the antisense oligomer of the present invention, and (ii) a multi-exon skipping experimental system using both the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention. The difference between the multi-exon skipping effect obtained in experimental system (ii) and the multi-exon skipping effect obtained in experimental system (i) can then be observed. 【0148】 The measurement of the multi-skipping effect is as described in section "1. Antisense Oligomers" of the present invention. 【0149】 The suppressor antisense oligomers of the present invention are 10-60 nucleotides long, 10-55 nucleotides long, 10-50 nucleotides long, 10-45 nucleotides long, 10-40 nucleotides long, 10-35 nucleotides long, 10-30 nucleotides long, 10-25 nucleotides long, 15-60 nucleotides long, 15-55 nucleotides long, 15-50 nucleotides long, 15-45 nucleotides long, 15-40 nucleotides long, 15-35 nucleotides long, 15-30 nucleotides long, 15-25 nucleotides long, 16-60 nucleotides long, 16-55 nucleotides long, 16-50 nucleotides long, 16-45 nucleotides long, 16-40 nucleotides long, 16-35 nucleotides long, 16-30 nucleotides long, 16-25 nucleotides long, 17- 60 base pairs in length, 17-55 base pairs, 17-50 base pairs, 17-45 base pairs, 17-40 base pairs, 17-35 base pairs, 17-30 base pairs, 17-25 base pairs, 18-60 base pairs, 18-55 base pairs, 18-50 base pairs, 18-45 base pairs, 18-40 base pairs, 18-35 base pairs, 18-30 base pairs, 18-25 base pairs, 19-60 base pairs, 19-55 base pairs, 19-50 base pairs, 19-45 base pairs, 19-40 base pairs, 19-35 base pairs, 19-30 base pairs, 19-25 base pairs, 20-60 base pairs, 20-55 base pairs, 20-50 base pairs The lengths are 20-45 nucleotides, 20-40 nucleotides, 20-35 nucleotides, 20-30 nucleotides, 20-25 nucleotides, 15-30 nucleotides, 15-29 nucleotides, 15-28 nucleotides, 15-27 nucleotides, 15-26 nucleotides, 15-25 nucleotides, 15-24 nucleotides, 15-23 nucleotides, 15-22 nucleotides, 15-21 nucleotides, 15-20 nucleotides, 15-19 nucleotides, 15-18 nucleotides, 16-30 nucleotides, 16-29 nucleotides, 16-28 nucleotides, 16-27 nucleotides, 16-26 nucleotides, 16-25 nucleotides, 16-24 nucleotides, 16-23 nucleotides, and 16-22 nucleotides. , 16-21 nucleotides long, 16-20 nucleotides long, 16-19 nucleotides long, 16-18 nucleotides long, 17-30 nucleotides long, 17-29 nucleotides long, 17-28 nucleotides long, 17-27 nucleotides long, 17-26 nucleotides long, 17-25 nucleotides long, 17-24 nucleotides long, 17-23 nucleotides long, 17-22 nucleotides long, 17-21 nucleotides long, 17-20 nucleotides long, 17-19 nucleotides long, 17-18 nucleotides long, 18-30 nucleotides long, 18-29 nucleotides long, 18-28 nucleotides long, 18-27 nucleotides long, 18-26 nucleotides long, 18-25 nucleotides long, 18-24 nucleotides long, 18-23 nucleotides long, 18-22 nucleotides long, 18-21 nucleotides long,They are 18-20 nucleotides long, 18-19 nucleotides long, 19-30 nucleotides long, 19-29 nucleotides long, 19-28 nucleotides long, 19-27 nucleotides long, 19-26 nucleotides long, 19-25 nucleotides long, 19-24 nucleotides long, 19-23 nucleotides long, 19-22 nucleotides long, 19-21 nucleotides long, 19-20 nucleotides long, 20-30 nucleotides long, 20-29 nucleotides long, 20- 28 nucleotides long, 20-27 nucleotides long, 20-26 nucleotides long, 20-25 nucleotides long, 20-24 nucleotides long, 20-23 nucleotides long, 20-22 nucleotides long, 20-21 nucleotides long, 60 nucleotides long, 59 nucleotides long, 58 nucleotides long, 57 nucleotides long, 56 nucleotides long, 55 nucleotides long, 54 nucleotides long, 53 nucleotides long, 52 nucleotides long, 51 nucleotides long, 50 nucleotides long, 49 nucleotides long, 48 salt The base length may be 47 nucleotides, 46 nucleotides, 45 nucleotides, 44 nucleotides, 43 nucleotides, 42 nucleotides, 41 nucleotides, 40 nucleotides, 39 nucleotides, 38 nucleotides, 37 nucleotides, 36 nucleotides, 35 nucleotides, 34 nucleotides, 33 nucleotides, 32 nucleotides, 31 nucleotides, 30 nucleotides, 29 nucleotides, 28 nucleotides, 27 nucleotides, 26 nucleotides, 25 nucleotides, 24 nucleotides, 23 nucleotides, 22 nucleotides, 21 nucleotides, 20 nucleotides, 19 nucleotides, 18 nucleotides, 17 nucleotides, 16 nucleotides, 15 nucleotides, 14 nucleotides, 13 nucleotides, 12 nucleotides, 11 nucleotides, or 10 nucleotides, but is not limited to these lengths, and the above lengths may be increased or decreased by 1, 2, or 3 nucleotides. 【0150】 The suppressor antisense oligomer of the present invention may be a linked antisense oligomer having a structure in which a plurality of unit oligomers are linked together, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. As described above, the unit oligomers may be linked via a linker that does not contribute to hybridization, or they may be linked directly without a linker. When the suppressor antisense oligomer of the present invention is linked, the base sequences of each unit oligomer are not continuous or overlapping with each other. 【0151】 The suppressor antisense oligomer of the present invention may be an oligonucleotide, a morpholino oligomer, or a peptide nucleic acid (PNA) oligomer. 【0152】 When the suppressor antisense oligomer of the present invention is an oligonucleotide (hereinafter referred to as "the suppressor antisense oligonucleotide of the present invention"), the suppressor antisense oligonucleotide of the present invention is a suppressor antisense oligomer having a nucleotide as its constituent unit, and such nucleotide may be a ribonucleotide, a deoxyribonucleotide, or a modified nucleotide. 【0153】 Modified nucleotides are as described above. 【0154】 The suppressor antisense oligonucleotide of the present invention is preferably a suppressor antisense oligomer of the present invention whose constituent unit is a group represented by the following general formula, wherein the -OH group at the 2' position of ribose is substituted with methoxy, and the phosphate bond is a phosphorothioate bond. [ka] (In the formula, Base has the same meaning as above.) 【0155】 The suppressor antisense oligonucleotides of the present invention can be easily synthesized using various automated synthesis equipment (e.g., AKTA oligopilot plus 10 / 100 (GE Healthcare)), or they can be manufactured by a third-party organization (e.g., Promega or Takara). 【0156】 When the suppressor antisense oligomer of the present invention is a morpholino oligomer (hereinafter referred to as "the suppressor antisense morpholino oligomer of the present invention"), the suppressor antisense morpholino oligomer of the present invention is a suppressor antisense oligomer having a group represented by the following general formula as its constituent unit. [ka] (In the formula, Base and W are the same as above.) 【0157】 Examples of morpholino monomer compounds used to synthesize the suppressor antisense morpholino oligomer of the present invention include, but are not limited to, the morpholino monomer compounds (A), (C), (T), and (G) shown in Table 7. 【0158】 [Table 7] 【0159】 In the suppressor antisense morpholino oligomer of the present invention, the morpholino oligomer is preferably PMO. 【0160】 Morpholino oligomers can be manufactured, for example, according to the methods described in International Publication No. 1991 / 009033 or International Publication No. 2009 / 064471. In particular, PMOs can be manufactured according to the methods described in International Publication No. 2009 / 064471 or International Publication No. 2013 / 100190. 【0161】 When the suppressor antisense oligomer of the present invention is a peptide nucleic acid oligomer (hereinafter referred to as "the suppressor antisense peptide nucleic acid oligomer of the present invention"), the suppressor antisense peptide nucleic acid oligomer of the present invention is a suppressor antisense oligomer having a group represented by the following general formula as its constituent unit. [ka] (In the formula, Base has the same meaning as above.) Peptide nucleic acids can be manufactured as described above. 【0162】 The suppressor antisense oligomer of the present invention may be in the form of a pharmaceutically acceptable salt, a hydrate, or a pharmaceutically acceptable salt hydrate. 【0163】 Examples of pharmaceutically acceptable salts of the suppressor antisense oligomer of the present invention include alkali metal salts such as sodium salts, potassium salts, and lithium salts; alkaline earth metal salts such as calcium salts and magnesium salts; metal salts such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, and cobalt salts; ammonium salts; t-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, and N-benzylphenethylamine salts. Examples include organic amine salts such as piperazine salts, tetramethylammonium salts, and tris(hydroxymethyl)aminomethane salt; hydrohalides such as hydrofluoric acid, hydrochloride, hydrobromide, and hydroiodide; inorganic salts such as nitrates, perchlorates, sulfates, and phosphates; lower alkanesulfonates such as methanesulfonates, trifluoromethanesulfonates, and ethanesulfonates; arylsulfonates such as benzenesulfonates and p-toluenesulfonates; organic salts such as acetates, malates, fumarates, succinates, citrates, tartrates, oxalates, and maleates; and amino acid salts such as glycine salts, lysine salts, arginine salts, ornithine salts, glutamates, and aspartates. These salts can be produced by known methods. Alternatively, the suppressor antisense oligomer of the present invention may also be in the form of its hydrate. 【0164】 [PMO manufacturing method] The antisense oligomer and the suppressor antisense oligomer of the present invention may be PMOs. One embodiment of a PMO is, for example, a compound represented by the following general formula (I) (hereinafter referred to as PMO(I)). [ka] (In the formula, each Base, R 2 , R 3This is synonymous with the above; n is any integer between 1 and 99, preferably any integer between 18 and 28. 【0165】 PMO(I) can be manufactured by known methods (see, for example, International Publication No. 2009 / 064471 or International Publication No. 2013 / 100190). 【0166】 Furthermore, the antisense oligomer or suppressor antisense oligomer of the present invention may have a 5' end that is any of the following chemical formulas (1) to (3). Preferably, it is (3)-OH. [ka] Hereinafter, the groups indicated in (1), (2), and (3) above will be referred to as "group (1)", "group (2)", and "group (3)", respectively. 【0167】 3. Pharmaceutical Uses The present invention provides a pharmaceutical composition (hereinafter referred to as "the pharmaceutical composition of the present invention") comprising the antisense oligomer of the present invention (including pharmaceutically acceptable salts thereof or hydrates thereof). The pharmaceutical composition of the present invention may further comprise the suppressor antisense oligomer of the present invention (including pharmaceutically acceptable salts thereof or hydrates thereof) and / or a pharmaceutically acceptable carrier. 【0168】 Furthermore, the present invention provides a pharmaceutical composition (hereinafter referred to as "the combination pharmaceutical of the present invention") comprising the antisense oligomer of the present invention (including its pharmaceutically acceptable salts or hydrates thereof) and the suppressor antisense oligomer of the present invention (including its pharmaceutically acceptable salts or hydrates thereof). 【0169】 When the pharmaceutical composition of the present invention contains the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention, any combination of these oligomers is included. Furthermore, in the combination pharmaceutical of the present invention, any combination of the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention is included. The antisense oligomer included in the pharmaceutical composition of the present invention or the combination pharmaceutical of the present invention may be any antisense oligomer of the present invention and is not particularly limited, but preferably it is selected from any one base sequence from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232. The antisense oligomer is, more preferably, SEQ ID NOs: 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 12 An antisense oligomer consisting of any one base sequence selected from the group consisting of 1, 122, 124, 125, 126, 128, 130, 131, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232, and more preferably the sequence It is an antisense oligomer consisting of any one base sequence selected from the group consisting of numbers 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228. Furthermore, the suppressor antisense oligomer contained in the pharmaceutical composition or combination pharmaceutical of the present invention may be any of the suppressor antisense oligomers of the present invention and is not particularly limited, but is preferably a suppressor antisense oligomer consisting of any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 257 to 275, more preferably SEQ ID NOs: 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, and more preferably a suppressor antisense oligomer consisting of any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 260, 261, and 263. When the pharmaceutical composition of the present invention contains the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention, it preferably contains a combination of the above-mentioned antisense oligomer and the suppressor antisense oligomer of the present invention. Furthermore, the combination pharmaceutical of the present invention preferably contains a combination of the above-mentioned antisense oligomer and the suppressor antisense oligomer of the present invention. 【0170】 Examples of combinations of the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention in the combination pharmaceutical of the present invention include the following. (K1) A combination in which the antisense oligomer is the oligomer corresponding to Sequence ID No. 75, and the suppressor antisense oligomer is the oligomer corresponding to Sequence ID No. 260. (K2) A combination in which the antisense oligomer is the oligomer corresponding to SEQ ID NO: 75, and the suppressor antisense oligomer is the oligomer corresponding to SEQ ID NO: 261. (K3) A combination in which the antisense oligomer is the oligomer corresponding to Sequence ID No. 75, and the suppressor antisense oligomer is the oligomer corresponding to Sequence ID No. 263. (K4) A combination in which the antisense oligomer is an oligomer consisting of SEQ ID NO: 65, and the suppressor antisense oligomer is an oligomer consisting of SEQ ID NO: 260. (K5) A combination in which the antisense oligomer is the oligomer corresponding to Sequence ID No. 65, and the suppressor antisense oligomer is the oligomer corresponding to Sequence ID No. 261. (K6) A combination in which the antisense oligomer is the oligomer corresponding to Sequence ID No. 65, and the suppressor antisense oligomer is the oligomer corresponding to Sequence ID No. 263. (K7) The antisense oligomer is a combination of oligomers consisting of SEQ ID NOs. 55 and 59, and the suppressor antisense oligomer is a combination of oligomers consisting of SEQ ID NO. 260. (K8) The antisense oligomer is a combination of oligomers consisting of SEQ ID NOs. 55 and 59, and the suppressor antisense oligomer is a combination of oligomers consisting of SEQ ID NO. 261. (K9) The antisense oligomer is a combination of oligomers consisting of sequence numbers 55 and 59, and the suppressor antisense oligomer is a combination of oligomers consisting of sequence number 263. The pharmaceutical composition of the present invention may include the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention in the above combination. 【0171】 The pharmaceutical compositions and combination pharmaceuticals of the present invention can be used, for example, for the treatment of Duchenne muscular dystrophy, Becker muscular dystrophy, limb-girdle muscular dystrophy (LGMD), congenital muscular dystrophy, Emery-Dreifuss muscular dystrophy, facioscapulohumeral muscular dystrophy, cancerous pharyngeal muscular dystrophy, autosomal dominant cerebral arteriovenous disease with subcortical infarction and leukoencephalopathy (CADASIL), Alport syndrome, and the like. Furthermore, the combination pharmaceuticals and pharmaceutical compositions of the present invention can be administered to human patients, particularly human patients with muscular dystrophy. Patients who are candidates for administration of the combination pharmaceuticals or pharmaceutical compositions of the present invention may be human patients who have mutations in the dystrophin gene that are subject to skipping of two or more exons selected from the group consisting of exons 45-55. 【0172】 In one aspect of the present invention, a method for treating muscular dystrophy is provided, comprising the step of administering the antisense oligomer of the present invention, or a combination of the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention, to a patient with muscular dystrophy. In another embodiment of the present invention, a method for treating muscular dystrophy is provided, comprising the step of administering a pharmaceutical composition or combination pharmaceutical of the present invention to a patient with muscular dystrophy. The aforementioned treatment method may involve skipping two or more consecutive exons selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA in numerical order. In the aforementioned treatment method, the muscular dystrophy patient may be a patient who has a mutation in the dystrophin gene that is subject to exon 45-55 skipping. The patient may be a human, or a human patient who has a mutation in the dystrophin gene that is subject to exon 45-55 skipping. 【0173】 The present invention further provides the use of the antisense oligomer of the present invention, or a combination of the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention, or the pharmaceutical composition of the present invention, or the combination pharmaceutical of the present invention, in the manufacture of pharmaceuticals for the treatment of muscular dystrophy. 【0174】 Furthermore, the present invention provides an antisense oligomer of the present invention for use in the treatment of muscular dystrophy, or a combination of the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention, or a pharmaceutical composition of the present invention, or a combination pharmaceutical of the present invention. The treatment may involve skipping two or more consecutive exons selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA. In the treatment, the muscular dystrophy patient may be a patient who has a mutation in the dystrophin gene that is subject to exon 45-55 skipping. The patient may be a human, or a human patient who has a mutation in the dystrophin gene that is subject to exon 45-55 skipping. 【0175】 The administration form of the antisense oligomer of the present invention, or a combination of the antisense oligomer of the present invention and the suppressor antisense oligomer of the present invention, or the pharmaceutical composition of the present invention, or the combination pharmaceutical of the present invention, is not particularly limited as long as it is a pharmaceutically acceptable administration form and can be selected according to the treatment method. However, from the viewpoint of ease of delivery to muscle tissue, intravenous administration, intra-arterial administration, intramuscular administration, subcutaneous administration, oral administration, intratissue administration, transdermal administration, etc., are preferred. Furthermore, the dosage forms that the composition of the present invention may take are not particularly limited, but examples include various injectable preparations, oral preparations, intravenous infusions, inhalants, ointments, lotions, etc. 【0176】 When administering the antisense oligomer and / or suppressor antisense oligomer of the present invention to a patient with muscular dystrophy, the composition of the present invention preferably includes a carrier that facilitates the delivery of the oligomer to muscle tissue. Such a carrier is not particularly limited as long as it is pharmaceutically acceptable, and examples include cationic carriers such as cationic liposomes and cationic polymers, or carriers utilizing a viral envelope. Examples of cationic liposomes include liposomes formed with 2-O-(2-diethylaminoethyl)carbamoyl-1,3-O-dioleoylglycerol and phospholipids as essential components (hereinafter referred to as "liposome A"), Oligofectamine® (manufactured by Invitrogen), Lipofecttin® (manufactured by Invitrogen), Lipofectamine® (manufactured by Invitrogen), Lipofectamine 2000® (manufactured by Invitrogen), DMRIE-C® (manufactured by Invitrogen), GeneSilencer® (manufactured by Gene Therapy Systems), TransMessenger® (manufactured by QIAGEN), TransIT TKO® (manufactured by Mirus), and Nucleofector II (Lonza). Among these, liposome A is preferred. Examples of cationic polymers include JetSI® (manufactured by Qbiogene) and Jet-PEI® (manufactured by Qbiogene). Examples of carriers utilizing the viral envelope include GenomeOne® (manufactured by Ishihara Sangyo Co., Ltd.) (HVJ-E liposome). Alternatively, the pharmaceutical device described in Patent No. 2924179, the republished patent application No. 2006 / 129594, and the republished patent application No. 2008 / 096690 can also be used. 【0177】 The concentration of the antisense oligomer of the present invention contained in the pharmaceutical composition and / or combination pharmaceutical of the present invention varies depending on the type of carrier, etc., but is suitable in the range of 0.1 nM to 100 μM, preferably in the range of 1 nM to 10 μM, and more preferably in the range of 10 nM to 1 μM. Furthermore, the weight ratio of the antisense oligomer of the present invention to the carrier (carrier / antisense oligomer of the present invention) contained in the composition of the present invention varies depending on the properties of the oligomer and the type of carrier, etc., but is suitable in the range of 0.1 to 100, preferably in the range of 1 to 50, and more preferably in the range of 10 to 20. The concentration of the suppressor antisense oligomer of the present invention contained in the pharmaceutical composition and / or combination pharmaceutical of the present invention varies depending on the type of carrier, etc., but is suitable in the range of 0.1 nM to 100 μM, preferably in the range of 1 nM to 10 μM, and more preferably in the range of 10 nM to 1 μM. Furthermore, the weight ratio of the suppressor antisense oligomer of the present invention to the carrier (carrier / suppressor antisense oligomer of the present invention) contained in the composition of the present invention varies depending on the properties of the oligomer and the type of carrier, etc., but is suitable in the range of 0.1 to 100, preferably in the range of 1 to 50, and more preferably in the range of 10 to 20. 【0178】 The pharmaceutical composition and / or combination pharmaceutical of the present invention may be in the form of an aqueous solution. In that case, the pharmaceutical composition and / or combination pharmaceutical of the present invention may contain the antisense oligomer of the present invention at concentrations of 2.5-500 mg / mL, 5-450 mg / mL, 10-400 mg / mL, 15-350 mg / mL, 20-300 mg / mL, 20-250 mg / mL, 20-200 mg / mL, 20-150 mg / mL, 20-100 mg / mL, 20-50 mg / mL, 20-40 mg / mL, 20-30 mg / mL, 23-27 mg / mL, 24-26 mg / mL, or 25 mg / mL. Alternatively, the pharmaceutical composition and / or combination pharmaceutical of the present invention may contain the antisense oligomer of the present invention at concentrations of 10-100 mg / mL, 15-95 mg / mL, 20-80 mg / mL, 25-75 mg / mL, 30-70 mg / mL, 35-65 mg / mL, 40-60 mg / mL, 45-55 mg / mL, 47-53 mg / mL, 48-52 mg / mL, 49-51 mg / mL, or 50 mg / mL. 【0179】 Furthermore, if the pharmaceutical composition and / or combination pharmaceutical of the present invention is in the form of an aqueous solution, the suppressor oligomer of the present invention may be included in concentrations of 2.5-500 mg / mL, 5-450 mg / mL, 10-400 mg / mL, 15-350 mg / mL, 20-300 mg / mL, 20-250 mg / mL, 20-200 mg / mL, 20-150 mg / mL, 20-100 mg / mL, 20-50 mg / mL, 20-40 mg / mL, 20-30 mg / mL, 23-27 mg / mL, 24-26 mg / mL, or 25 mg / mL. Alternatively, the pharmaceutical composition and / or combination pharmaceutical of the present invention may contain the suppressor oligomer of the present invention at concentrations of 10-100 mg / mL, 15-95 mg / mL, 20-80 mg / mL, 25-75 mg / mL, 30-70 mg / mL, 35-65 mg / mL, 40-60 mg / mL, 45-55 mg / mL, 47-53 mg / mL, 48-52 mg / mL, 49-51 mg / mL, or 50 mg / mL. 【0180】 The pharmaceutical composition and / or combination pharmaceutical of the present invention may be in dry form. In that case, to prepare the pharmaceutical composition and / or combination pharmaceutical of the present invention in aqueous solution form, for example, 125 mg or 250 mg of the antisense oligomer of the present invention in dry form may be mixed with 0.5 mL to 100 mL of water (corresponding to an antisense oligomer concentration of the present invention of 1.25 mg / mL to 250 mg / mL or 2.5 mg / mL to 500 mg / mL), preferably with 1 mL to 50 mL of water (corresponding to an antisense oligomer concentration of the present invention of 2.5 mg / mL to 125 mg / mL or 5 mg / mL to 250 mg / mL), and more preferably with 5 mL to 10 mL of water (corresponding to an antisense oligomer concentration of the present invention of 12.5 mg / mL to 25 mg / mL or 25 mg / mL to 50 mg / mL). 【0181】 Furthermore, if the pharmaceutical composition and / or combination pharmaceutical of the present invention is in a dry form, in order to prepare the pharmaceutical composition and / or combination pharmaceutical of the present invention in aqueous solution form, for example, 125 mg or 250 mg of the suppressor oligomer of the present invention in dry form may be mixed with 0.5 mL to 100 mL of water (corresponding to a suppressor oligomer concentration of the present invention of 1.25 mg / mL to 250 mg / mL or 2.5 mg / mL to 500 mg / mL), preferably with 1 mL to 50 mL of water (corresponding to a suppressor oligomer concentration of the present invention of 2.5 mg / mL to 125 mg / mL or 5 mg / mL to 250 mg / mL), and more preferably with 5 mL to 10 mL of water (corresponding to a suppressor oligomer concentration of the present invention of 12.5 mg / mL to 25 mg / mL or 25 mg / mL to 50 mg / mL). 【0182】 The concentrations of the antisense oligomer and the suppressor oligomer of the present invention contained in the pharmaceutical composition and / or combination pharmaceutical of the present invention may be the concentrations of the antisense oligomer and the suppressor oligomer individually, or the combined concentration of the antisense oligomer and the suppressor oligomer of the present invention. 【0183】 The pharmaceutical composition and / or combination pharmaceutical of the present invention may optionally contain pharmaceutically acceptable additives in addition to the antisense oligomer and / or suppressor antisense oligomer of the present invention and the carrier described above. Examples of such additives include emulsifying agents (e.g., fatty acids having 6 to 22 carbon atoms and their pharmaceutically acceptable salts, albumin, dextran), stabilizers (e.g., cholesterol, phosphatidic acid, mannitol, sorbitol), isotonic agents (e.g., sodium chloride, glucose, maltose, lactose, sucrose, trehalose), and pH adjusters (e.g., hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, sodium hydroxide, potassium hydroxide, triethanolamine). One or more of these may be used. The content of such additives in the composition of the present invention is appropriately 90% by weight or less, preferably 70% by weight or less, and more preferably 50% by weight or less. 【0184】 The pharmaceutical composition and / or combination pharmaceutical of the present invention can be prepared by adding the antisense oligomer and / or suppressor antisense oligomer of the present invention to a dispersion of a carrier and stirring appropriately. The additive can also be added at an appropriate step, either before or after the addition of the antisense oligomer and / or suppressor antisense oligomer of the present invention. The aqueous solvent that can be used when adding the antisense oligomer and / or suppressor antisense oligomer of the present invention is not particularly limited as long as it is pharmaceutically acceptable. Examples include electrolyte solutions such as water for injection, distilled water for injection, and physiological saline, as well as sugar solutions such as glucose solution and maltose solution. Furthermore, the pH, temperature, and other conditions in such cases can be appropriately selected by those skilled in the art. 【0185】 The pharmaceutical composition and / or combination pharmaceutical of the present invention can be, for example, a liquid formulation or a lyophilized formulation thereof. The lyophilized formulation can be prepared by conventional methods by lyophilizing the composition of the present invention, which is in the form of a liquid formulation. For example, after appropriate sterilization of the composition of the present invention, which is in the form of a liquid formulation, a predetermined amount can be dispensed into a vial, pre-freezed for about 2 hours under conditions in the range of approximately -40°C to -20°C, primary drying can be performed under reduced pressure in the range of approximately 0°C to 10°C, and then secondary drying can be performed under reduced pressure in the range of approximately 15°C to 25°C to lyophilize the composition. Generally, the inside of the vial can then be replaced with nitrogen gas and the vial can be capped to obtain a lyophilized formulation of the composition of the present invention. 【0186】 The lyophilized preparations of the pharmaceutical composition and / or combination pharmaceutical of the present invention can generally be redissolved and used by adding any suitable solution (redissolving solution). Examples of such redissolving solutions include water for injection, physiological saline, and other general intravenous fluids. The volume of this redissolving solution varies depending on the application and is not particularly limited, but 0.5 to 2 times the volume before lyophilization or 500 mL or less is appropriate. 【0187】 When administering the pharmaceutical composition and / or combination pharmaceutical of the present invention, it is desirable to adjust the dosage based on the type and dosage form of the antisense oligomer and / or suppressor antisense oligomer contained in the present invention, the patient's condition such as age and weight, the route of administration, and the nature and severity of the disease. However, for adults, the amount of the antisense oligomer and / or suppressor antisense oligomer of the present invention can be administered in a single dose of 0.1 mg to 1 g per kg of body weight, preferably 1 mg to 100 mg per kg of body weight, more preferably 1 mg to 90 mg per kg of body weight, and more preferably 1 mg to 80 mg per kg of body weight. The frequency of administration may be once every 1 to 3 days, once a week, or once every 2 to 3 weeks. These values ​​may also vary depending on the type of disease being targeted, the form of administration, and the target molecule. Therefore, in some cases, a lower dose or frequency of administration may be sufficient, and conversely, there may be times when a higher dose or frequency of administration is necessary. 【0188】 Another embodiment of the pharmaceutical composition and / or combination pharmaceutical of the present invention is a pharmaceutical composition comprising a vector capable of expressing the antisense oligomer and / or suppressor antisense oligomer of the present invention and the carrier described above. Such an expression vector may be capable of expressing multiple antisense oligomers and / or suppressor antisense oligomers of the present invention. Similar to the composition of the present invention containing the antisense oligomer and / or suppressor antisense oligomer of the present invention, pharmaceutically acceptable additives may be added to the composition. The concentration of the expression vector contained in the composition varies depending on the type of carrier, etc., but is suitable in the range of 0.1 nM to 100 μM, preferably in the range of 1 nM to 10 μM, and more preferably in the range of 10 nM to 1 μM. The weight ratio of the expression vector to the carrier (carrier / expression vector) contained in the composition varies depending on the properties of the expression vector, the type of carrier, etc., but is suitable in the range of 0.1 to 100, preferably in the range of 1 to 50, and more preferably in the range of 10 to 20. Furthermore, the amount of carrier contained in the composition is the same as in the case of the composition of the present invention containing the antisense oligomer and / or the suppressor antisense oligomer of the present invention, and the preparation method and the like are also the same as in the case of the composition of the present invention. 【0189】 4. Pre-mRNAs other than human dystrophin This specification specifically describes the case where the target pre-mRNA is human dystrophin pre-mRNA, but is not limited thereto. For target pre-mRNAs other than human dystrophin, the design, manufacture, and use of antisense oligomers and suppressor antisense oligomers can be carried out in accordance with the disclosures herein or known art and methods. Those skilled in the art can design, manufacture, and use antisense oligomers or suppressor antisense oligomers for target pre-mRNAs other than human dystrophin based on the disclosures herein or known art and methods. Similarly, those skilled in the art can perform multi-exon skipping of target pre-mRNA using antisense oligomers and enhance the efficiency of multi-exon skipping using suppressor antisense oligomers based on the disclosures herein and known art and methods. 【0190】 In this specification, examples of target pre-mRNAs include human γ-sarcoglycan (SGCG), human Notch3, and human dystrophin pre-mRNAs. In one embodiment of the present invention, the target pre-mRNA may be human dystrophin pre-mRNA. In another embodiment of the present invention, the target pre-mRNA may be SGCG or human Notch3. 【0191】 The nucleotide sequences of the target regions of the antisense oligomers used for multi-exon skipping in target pre-mRNAs of human SGCG and human Notch3 are as follows. 【0192】 In the present invention, the target regions of human SGCG pre-mRNA are shown below as Sequence IDs 283 and 284. 【0193】 [Table 8] 【0194】 The target region of human Notch3 pre-mRNA in this invention is shown below. 【0195】 [Table 9] 【0196】 Antisense oligomers for human SGCG that induce multi-exon skipping of human SGCG pre-mRNA include, for example, those that target the range of -400 to +50 bases starting from the 3' end of intron 3. A specific example is: (L1) Any one base sequence selected from the group consisting of sequence numbers 287-308, or (L2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±15% of the length of the selected nucleotide sequence. Examples include antisense oligomers for human SGCG consisting of the above, or pharmaceutically acceptable salts thereof, or hydrates thereof. 【0197】 Here, the base sequence of (L2) is a variant of the base sequence of (L1), but as a variant, (L2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±15% of the length of the selected nucleotide sequence. (L2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±14% of the length of the selected nucleotide sequence. (L2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±13% of the length of the selected nucleotide sequence. (L2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±12% of the length of the selected nucleotide sequence. (L2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±11% of the length of the selected nucleotide sequence. (L2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±10% of the length of the selected nucleotide sequence. (L2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±9% of the length of the selected nucleotide sequence. (L2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±8% of the length of the selected nucleotide sequence. (L2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±7% of the length of the selected nucleotide sequence. (L2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±6% of the length of the selected nucleotide sequence. (L2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±5% of the length of the selected nucleotide sequence. (L2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±4% of the length of the selected nucleotide sequence. (L2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±3% of the length of the selected nucleotide sequence. (L2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±2% of the length of the selected nucleotide sequence. (L2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±1% of the length of the selected nucleotide sequence, and (L2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 287 to 308, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0198】 Therefore, further embodiments of antisense oligomers for human SGCG include: Examples include antisense oligomers for human SGCG consisting of any one base sequence selected from the group consisting of (L1) and (L2-1) to (L2-16), or pharmaceutically acceptable salts thereof, or hydrates thereof. Antisense oligomers for human SGCG are included in the present invention. 【0199】 Examples of suppressor-antisense oligomers for human SGCG pre-mRNA single skipping (suppressor-antisense oligomers for human SGCG) include those targeting the intron 4 splicing silencer sequence or the single-skipping splice site, and specific examples include: (M1) Any one base sequence selected from the group consisting of sequence numbers 331-335, or (M2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±15% of the length of the selected nucleotide sequence. Examples include human SGCG suppressor antisense oligomers or pharmaceutically acceptable salts thereof, or hydrates thereof. 【0200】 Here, the base sequence of (M2) is a variant of the base sequence of (M1), but as a variant, (M2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±15% of the length of the selected nucleotide sequence. (M2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±14% of the length of the selected nucleotide sequence. (M2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±13% of the length of the selected nucleotide sequence. (M2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±12% of the length of the selected nucleotide sequence. (M2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±11% of the length of the selected nucleotide sequence. (M2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±10% of the length of the selected nucleotide sequence. (M2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±9% of the length of the selected nucleotide sequence. (M2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±8% of the length of the selected nucleotide sequence. (M2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±7% of the length of the selected nucleotide sequence. (M2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±6% of the length of the selected nucleotide sequence. (M2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±5% of the length of the selected nucleotide sequence. (M2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±4% of the length of the selected nucleotide sequence. (M2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±3% of the length of the selected nucleotide sequence. (M2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±2% of the length of the selected nucleotide sequence. (M2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±1% of the length of the selected nucleotide sequence, and (M2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 331 to 335, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0201】 Therefore, further embodiments of suppressor antisense oligomers for human SGCG include: Examples include human SGCG suppressor antisense oligomers consisting of any one base sequence selected from the group consisting of (M1) and (M2-1) to (M2-16), or pharmaceutically acceptable salts thereof, or hydrates thereof. Human SGCG suppressor antisense oligomers are included in the present invention. Human SGCG suppressor antisense oligomers are included in the present invention. Human SGCG suppressor antisense oligomers can increase the efficiency of multi-skipping of SGCG pre-mRNA. 【0202】 Examples of antisense oligomers for human Notch3 that induce multi-exon skipping of human Notch3 pre-mRNA include those that target the region near the acceptor of intron 3, i.e., the range of -400 to +50 base pairs starting from the 3' end of intron 3. (N1) Any one base sequence selected from the group consisting of sequence numbers 309-330, or (N2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±15% of the length of the selected nucleotide sequence. Examples include antisense oligomers for human Notch3 consisting of the above, or pharmaceutically acceptable salts thereof, or hydrates thereof. 【0203】 Here, the base sequence of (N2) is a variant of the base sequence of (N1), but as a variant, (N2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±15% of the length of the selected nucleotide sequence. (N2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±14% of the length of the selected nucleotide sequence. (N2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±13% of the length of the selected nucleotide sequence. (N2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±12% of the length of the selected nucleotide sequence. (N2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±11% of the length of the selected nucleotide sequence. (N2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±10% of the length of the selected nucleotide sequence. (N2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±9% of the length of the selected nucleotide sequence. (N2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±8% of the length of the selected nucleotide sequence. (N2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±7% of the length of the selected nucleotide sequence. (N2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±6% of the length of the selected nucleotide sequence. (N2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±5% of the length of the selected nucleotide sequence. (N2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±4% of the length of the selected nucleotide sequence. (N2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±3% of the length of the selected nucleotide sequence. (N2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±2% of the length of the selected nucleotide sequence. (N2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±1% of the length of the selected nucleotide sequence, and (N2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 309 to 330, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0204】 Therefore, further embodiments of the human Notch3 antisense oligomer include a human Notch3 antisense oligomer consisting of any one base sequence selected from the group consisting of (N1) and (N2-1) to (N2-16), or a pharmaceutically acceptable salt thereof, or a hydrate thereof. The human Notch3 antisense oligomer is included in the present invention. 【0205】 Examples of human Notch3 pre-mRNA single-skip suppressor antisense oligomers (human Notch3 suppressor antisense oligomers) include those targeting the intron 4 splice-sensing silencer sequence or the single-skip splice site, and specific examples include: (O1) Any one base sequence selected from the group consisting of sequence numbers 336-340, or (O2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±15% of the length of the selected nucleotide sequence. Examples include human Notch3 suppressor antisense oligomers or pharmaceutically acceptable salts thereof, or hydrates thereof. 【0206】 Here, the base sequence of (O2) is a variant of the base sequence of (O1), but as a variant, (O2-1) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±15% of the length of the selected nucleotide sequence. (O2-2) A nucleotide sequence having 86% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±14% of the length of the selected nucleotide sequence. (O2-3) A nucleotide sequence having 87% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±13% of the length of the selected nucleotide sequence. (O2-4) A nucleotide sequence having 88% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±12% of the length of the selected nucleotide sequence. (O2-5) A nucleotide sequence having 89% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±11% of the length of the selected nucleotide sequence. (O2-6) A nucleotide sequence having 90% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±10% of the length of the selected nucleotide sequence. (O2-7) A nucleotide sequence having 91% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±9% of the length of the selected nucleotide sequence. (O2-8) A nucleotide sequence having 92% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±8% of the length of the selected nucleotide sequence. (O2-9) A nucleotide sequence having 93% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±7% of the length of the selected nucleotide sequence. (O2-10) A nucleotide sequence having 94% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±6% of the length of the selected nucleotide sequence. (O2-11) A nucleotide sequence having 95% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±5% of the length of the selected nucleotide sequence. (O2-12) A nucleotide sequence having 96% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±4% of the length of the selected nucleotide sequence. (O2-13) A nucleotide sequence having 97% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±3% of the length of the selected nucleotide sequence. (O2-14) A nucleotide sequence having 98% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±2% of the length of the selected nucleotide sequence. (O2-15) A nucleotide sequence having 99% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±1% of the length of the selected nucleotide sequence, and (O2-16) A nucleotide sequence having 99.5% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 336 to 340, and having a length within ±0.5% of the length of the selected nucleotide sequence. This can also be mentioned. 【0207】 Therefore, further embodiments of the suppressor antisense oligomer for human Notch3 include: Examples include human Notch3 suppressor antisense oligomers consisting of any one base sequence selected from the group consisting of (O1) and (O2-1) to (O2-16), or pharmaceutically acceptable salts thereof, or hydrates thereof. Human Notch3 suppressor antisense oligomers are included in the present invention. Human Notch3 suppressor antisense oligomers can increase the efficiency of Notch3 pre-mRNA multi-skipping. 【0208】 The nucleotide sequences of sequence numbers 287-340 are shown below. TIFF2026095457000032.tif255165TIFF2026095457000033.tif42170 【0209】 5. Methods to increase the efficiency of multi-exon skipping The present invention includes a step of inhibiting a splicing silencer sequence, splice site sequence, or branch site sequence of a target pre-mRNA when skipping two or more sequentially numbered exons from the target pre-mRNA. The present invention provides a method for increasing the efficiency of skipping two or more consecutively numbered exons (hereinafter referred to as "the method for increasing efficiency of the present invention"). 【0210】 In the method for increasing the mRNA of the present invention, skipping of two or more sequentially numbered exons of the target pre-mRNA can be performed using an antisense oligomer that induces multi-exon skipping, such as the antisense oligomer of the present invention, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0211】 The present invention provides a method for increasing the efficiency of multi-exon skipping by inhibiting splicing silencer sequences, splice site sequences, or branch site sequences on target pre-mRNA, thereby suppressing single skipping. The present invention provides a method for increasing the efficiency of multi-exon skipping by inhibiting the function of these splicing silencer sequences, splice site sequences, or branch site sequences on target pre-mRNA, and more specifically, by targeting these sequences with suppressor antisense oligomers to suppress single skipping of exons. 【0212】 Furthermore, in the amplification method of the present invention, the step of inhibiting the splicing silencer sequence, splice site, or branch site sequence can be carried out using a suppressor antisense oligomer such as the suppressor antisense oligomer of the present invention, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 【0213】 In the amplification method of the present invention, an oligomer containing a nucleotide sequence complementary to the splicing silencer sequence, branch site sequence, and splice site sequence on the target pre-mRNA can be used as a suppressor antisense oligomer in the step of inhibiting the splicing silencer sequence, branch site sequence, and splice site sequence. 【0214】 The splicing silencer sequence, branch site sequence, and splice site sequence are as described above. In one embodiment of the augmentation method of the present invention, the splicing silencer sequence may be a recognition sequence for heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1). Examples of human hnRNP A1 recognition sequences are shown in SEQ ID NOs. 370-379. 【0215】 In the present invention, "using" the antisense oligomer and the suppressor antisense oligomer in the method of amplification means introducing the antisense oligomer and the suppressor antisense oligomer into cells expressing the target pre-mRNA, thereby causing exon skipping in the target pre-mRNA. Methods for introducing the antisense oligomer and the suppressor antisense oligomer into the cells include introduction methods using cationic carriers such as cationic liposomes and cationic polymers, or carriers utilizing the viral envelope. Examples of cationic liposomes include liposomes formed with 2-O-(2-diethylaminoethyl)carbamoyl-1,3-O-dioleoylglycerol and phospholipids as essential components (hereinafter referred to as "liposome A"), Oligofectamine® (manufactured by Invitrogen), Lipofecttin® (manufactured by Invitrogen), Lipofectamine® (manufactured by Invitrogen), Lipofectamine 2000® (manufactured by Invitrogen), DMRIE-C® (manufactured by Invitrogen), GeneSilencer® (manufactured by Gene Therapy Systems), TransMessenger® (manufactured by QIAGEN), TransIT TKO® (manufactured by Mirus), and Nucleofector II (Lonza). Among these, liposome A is preferred. Examples of cationic polymers include JetSI® (manufactured by Qbiogene) and Jet-PEI® (manufactured by Qbiogene). Examples of carriers utilizing the viral envelope include GenomeOne® (manufactured by Ishihara Sangyo Co., Ltd.) (HVJ-E liposome). Alternatively, the pharmaceutical device described in Patent No. 2924179, the republished patent application No. 2006 / 129594, and the republished patent application No. 2008 / 096690 can also be used. 【0216】 Furthermore, as a method for introducing the antisense oligomer and the suppressor antisense oligomer into cells expressing the target pre-mRNA in the patient's body, one can refer to the administration method described in item "3. Pharmaceutical Uses". 【0217】 In the augmentation method of the present invention, by inhibiting the splicing silencer sequence, splice site sequence, or branch site sequence of the target pre-mRNA, if the efficiency of multi-exon skipping obtained without such inhibition is set to 100, then the efficiency is 2% or more, 4% or more, 6% or more, 8% or more, 10% or more, 12% or more, 14% or more, 16% or more, 18% or more, 20% or more, 22% or more, 24% or more, 26% or more, 28% or more, 30% or more, 32% or more, 3 It increases by 4% or more, 36% or more, 38% or more, 40% or more, 42% or more, 44% or more, 46% or more, 48% or more, 50% or more, 52% or more, 54% or more, 56% or more, 58% or more, 60% or more, 62% or more, 64% or more, 66% or more, 68% or more, 70% or more, 72% or more, 74% or more, 76% or more, 78% or more, 80% or more, 82% or more, 84% or more, 86% or more, 88% or more, 90% or more, 92% or more, 94% or more, 96% or more, 98% or more, or 100% or more. 【0218】 In the amplification method of the present invention, the target pre-mRNA can be, for example, human γ-sarcoglycan (SGCG), human Notch3, and human dystrophin pre-mRNA. In one embodiment of the present invention, the target pre-mRNA is human dystrophin pre-mRNA. When the target pre-mRNA is human dystrophin pre-mRNA, the two or more sequentially numbered exons are selected from the group consisting of the 45th to the 55th exons of the human dystrophin pre-mRNA. [Examples] 【0219】 The present invention will be described in more detail below with reference to examples and test examples, but the present invention is not limited to the scope shown in the examples. 【0220】 [Example 1: Production of antisense oligomers] Preparation of 4-{[(2S,6R)-6-(4-benzamido-2-oxopyrimidine-1-yl)-4-tritylmorpholine-2-yl]methoxy}-4-oxobutanoic acid (Compound 1) supported on aminopolystyrene resin Step 1: Preparation of 4-{[(2S,6R)-6-(4-benzamido-2-oxopyrimidine-1(2H)-yl)-4-tritylmorpholine-2-yl]methoxy}-4-oxobutanoic acid Under an argon atmosphere, 3.44 g of N-{1-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl]-2-oxo-1,2-dihydropyrimidine-4-yl}benzamide and 1.1 g of 4-dimethylaminopyridine (4-DMAP) were suspended in 50 mL of dichloromethane, and 0.90 g of succinic anhydride was added. The mixture was stirred at room temperature for 3 hours. 10 mL of methanol was added to the reaction mixture, and the mixture was concentrated under reduced pressure. The residue was extracted using ethyl acetate and a 0.5 M aqueous potassium dihydrogen phosphate solution. The resulting organic layer was washed sequentially with a 0.5 M aqueous potassium dihydrogen phosphate solution, water, and saturated brine. The resulting organic layer was dried over sodium sulfate and concentrated under reduced pressure to obtain 4.0 g of the target product. Step 2: Production of 4-{[(2S,6R)-6-(4-benzamido-2-oxopyrimidine-1-yl)-4-tritylmorpholin-2-yl]methoxy}-4-oxobutanoic acid supported on aminopolystyrene resin 4.0 g of 4-{[(2S,6R)-6-(4-benzamido-2-oxopyrimidine-1(2H)-yl)-4-tritylmorpholine-2-yl]methoxy}4-oxobutanoic acid was dissolved in 200 mL of pyridine (anhydrous), and 0.73 g of 4-DMAP and 11.5 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride were added. Then, 25.0 g of aminopolystyrene resin: Primer support 200 amino (GE Healthcare Japan, 17-5214-97) and 8.5 mL of triethylamine were added, and the mixture was shaken at room temperature for 4 days. After the reaction, the resin was filtered off. The obtained resin was washed sequentially with pyridine, methanol, and dichloromethane, and dried under reduced pressure. 200 mL of tetrahydrofuran (anhydrous), 15 mL of acetic anhydride, and 15 mL of 2,6-lutidine were added to the obtained resin, and the mixture was shaken at room temperature for 2 hours. The resin was filtered, washed sequentially with pyridine, methanol, and dichloromethane, and dried under reduced pressure to obtain 26.7 g of the target product. The loading amount of the target substance was determined by measuring the UV absorbance at 409 nm for the molar amount of trityl per gram of resin using a known method. The loading amount of the resin was 129.2 μmol / g. 【0221】 UV measurement conditions Equipment: U-2910 (Hitachi, Ltd.) Solvent: Methanesulfonic acid Wavelength: 409 nm ε value: 45000 【0222】 Preparation of 4-{[(2S,6R)-6-(5-methyl-2,4-dioxopyrimidine-1-yl)-4-tritylmorpholin-2-yl]methoxy}-4-oxobutanoic acid (Compound 2) supported on aminopolystyrene resin The indicated compound was prepared in the same manner as for compound 1. However, instead of N-{1-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholine-2-yl]-2-oxo-1,2-dihydropyrimidine-4-yl}benzamide used in step 1 of the preparation of compound 1, 1-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholine-2-yl]-5-methylpyrimidine-2,4(1H,3H)-dione was used in this step. The loading amount of the target substance was determined by measuring the UV absorbance at 409 nm for the molar amount of trityl per gram of resin using a known method. The loading amount of the resin was 164.0 μmol / g. 【0223】 Preparation of 4-{[(2S,6R)-6-(6-benzamidopurine-9-yl)-4-tritylmorpholin-2-yl]methoxy}-4-oxobutanoic acid (compound 3) supported on aminopolystyrene resin The indicated compound was prepared in the same manner as for compound 1. However, instead of N-{1-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholine-2-yl]-2-oxo-1,2-dihydropyrimidine-4-yl}benzamide used in step 1 of the preparation of compound 1, N-{9-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholine-2-yl]purine-6-yl}benzamide was used in this step. The loading amount of the target substance was determined by measuring the UV absorbance at 409 nm for the molar amount of trityl per gram of resin using a known method. The loading amount of the resin was 185.7 μmol / g. 【0224】 Preparation of 4-{{(2S,6R)-6-{6-(2-cyanoethoxy)-2-[(2-phenoxyacetyl)amino]purine-9-yl}-4-tritylmorpholin-2-yl}methoxy}-4-oxobutanoic acid (compound 4) supported on aminopolystyrene resin The indicated compound was prepared in the same manner as for compound 1. However, instead of N-{1-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholine-2-yl]-2-oxo-1,2-dihydropyrimidine-4-yl}benzamide used in step 1 of the preparation of compound 1, N-{6-(2-cyanoethoxy)-9-[(2R,6S)-6-(hydroxymethyl)-4-tritylmorpholine-2-yl]purine-2-yl}-2-phenoxyacetamide was used in this step. The loading amount of the target substance was determined by measuring the UV absorbance at 409 nm for the molar amount of trityl per gram of resin using a known method. The loading amount of the resin was 164.8 μmol / g. 【0225】 Production of antisense PMOs targeting donor or acceptor regions of the introns of the human dystrophin gene. Following the instructions below, PMOs with the nucleotide sequences (SEQ ID NOs. 1-232) of PMO No. 1-232 in Table 11 were synthesized, targeting the region near the donor or acceptor introns 44-55 of human dystrophin pre-mRNA shown in Table 10. The 5' end of the PMO is the group (3) described below. The synthesized PMOs were dissolved in sterile water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.). [ka] 【0226】 The target nucleotide sequences of the antisense oligomer or suppressor antisense oligomer of the present invention are described as follows: "Ha1_b1-c1", "Ha2_b2-c2_Ha3_b3-c3", or "Ha4_b4-c4_Ha5_b5-c5_Ha6_b6-c6". "Ha1_b1-c1" is the target nucleotide sequence of the antisense oligomer according to the first embodiment of this specification. "Ha1" represents the a-th exon of the human dystrophin gene, "b1" represents the 5' end of the target nucleotide sequence, and "c1" represents the 3' end of the target nucleotide sequence. "Ha1_b1-c1" may have an additional nucleotide at its 3' end, and such a target nucleotide sequence is represented as "Ha1_b1-c1_N" (where "N" represents any nucleotide). When "b1" and "c1" are positive integers, they represent the position of the base in the 3' direction relative to the 5' end of the a-th exon, with the 5' end base being the 1st base, respectively. On the other hand, when "b1" and "c1" are negative integers, they represent the position of the base in the 5' direction relative to the (a-1)-th intron, with the 3' end base being the -1st base, respectively. For example, "H45_(-10)-15" means a base sequence in which the 5' end of the target base sequence is the 10th base in the direction from the 3' end to the 5' end of the 44th intron, and the 3' end of the target base sequence is the 15th base in the direction from the 5' end to the 3' end of the 45th exon. "Ha2_b2-c2_Ha3_b3-c3" is the target nucleotide sequence of the antisense oligomer according to the second embodiment described herein. The first part of "Ha2_b2-c2_Ha3_b3-c3", "Ha2_b2-c2", means the target nucleotide sequence of the 3' terminal unit oligomer constituting the antisense oligomer, and the second part, "Ha3_b3-c3", means the target nucleotide sequence of the 5' terminal unit oligomer constituting the antisense oligomer. "Ha3_b3-c3" may have an additional base at its 3' end, and such a target nucleotide sequence is represented as "Ha2_b2-c2_Ha3_b3-c3_N" (where "N" represents any base). Each "Ha2_b2-c2" and "Ha3_b3_-c3" is represented in the same manner as "Ha1_b1-c1" above. Note that if "Ha2" and "Ha3" are the same, the "_Ha3" part can be omitted. For example, "H45_(-5)-5_25-35" or "H45_(-5)-5_H45_25-35" means a nucleotide sequence in which the target nucleotide sequence of the 3' terminal unit oligomer constituting the antisense oligomer is "H45_(-5)-5" and the target nucleotide sequence of the 5' terminal unit oligomer is "H45_25-35". "Ha4_b4-c4_Ha5_b5-c5_Ha6_b6-c6" is the target nucleotide sequence of the antisense oligomer in the third embodiment described herein. The first part of "Ha4_b4-c4_Ha5_b5-c5_Ha6_b6-c6", "Ha4_b4-c4", means the target nucleotide sequence of the 3' terminal unit oligomer constituting the antisense oligomer; the second part, "Ha5_b5-c5", means the target nucleotide sequence of the intermediate unit oligomer constituting the antisense oligomer; and the third part, "Ha6_b6-c6", means the target nucleotide sequence of the 5' terminal unit oligomer constituting the antisense oligomer. "Ha6_b6-c6" may have an additional base at its 3' end, and such a target base sequence is represented as "Ha4_b4-c4_Ha5_b5-c5_Ha6_b6-c6_N" (where "N" represents any base). Each "Ha4_b4-c4", "Ha5_b5_-c5", and "Ha6_b6_-c6" is represented in the same way as "Ha1_b1-c1" above. Note that if "Ha4", "Ha5", and "Ha6" are the same, the "_Ha5" and "_Ha6" parts can be omitted. For example, "H45_(-5)-5_25-35_60-70" or "H45_(-5)-5_H45_25-35_H45_60-70" means that the target base sequence of the 3' terminal unit oligomer constituting the antisense oligomer is "H45_(-5)-5", the target base sequence of the intermediate unit oligomer is "H45_25-35", and the target base sequence of the 5' terminal unit oligomer is "H45_60-70". 【0227】 [Table 10] TIFF2026095457000036.tif254155 【0228】 [Table 11] TIFF2026095457000038.tif255163TIFF2026095457000039.tif255163TIFF2026095457 000040.tif255162TIFF2026095457000041.tif255165TIFF2026095457000042.tif46170 【0229】 Among the PMOs in Table 11, Examples of those targeting R2 in another embodiment include PMO Nos. 22-71, 91-93, and 98-99, and in yet another embodiment, PMO Nos. 22-59, 91-93, and 98-99; Examples of target regions with R4 include PMO Nos. 150-163 in another embodiment, and PMO Nos. 150-160 in yet another embodiment; Examples of those targeting R6 include PMO Nos. 171-177 in another embodiment, and PMO Nos. 171-176 in yet another embodiment; In another embodiment, PMO No. 180 targets R8; in another embodiment, PMO No. 183 targets R10; and in another embodiment, PMO Nos. 186-191 target R12; Examples of those targeting R16 include PMO Nos. 197 and 198 in another embodiment, and PMO No. 197 in yet another embodiment; In another embodiment, PMO Nos. 201-206 target the R18 region; Examples of those targeting R22 include PMO Nos. 217-223 in another embodiment, and PMO Nos. 217-222 in yet another embodiment; In another embodiment, PMO Nos. 128-142 target R24. 【0230】 4-{[(2S,6R)-6-(4-benzamido-2-oxopyrimidine-1-yl)-4-tritylmorpholine-2-yl]methoxy}-4-oxobutanoic acid (compound 1) supported on aminopolystyrene resin, corresponding to the 5' terminal base, 4-{[(2S,6R)-6-(5-methyl-2,4-dioxopyrimidine-1-yl)-4-tritylmorpholine-2-yl]methoxy}-4-oxobutanoic acid (compound 2) supported on aminopolystyrene resin, supported on aminopolystyrene resin 0.2 g of either 4-{[(2S,6R)-6-(6-benzamidepurine-9-yl)-4-tritylmorpholine-2-yl]methoxy}-4-oxobutanoic acid (compound 3) or 4-{{(2S,6R)-6-{6-(2-cyanoethoxy)-2-[(2-phenoxyacetyl)amino]purine-9-yl}-4-tritylmorpholine-2-yl}methoxy}-4-oxobutanoic acid (compound 4) supported on an aminopolystyrene resin was packed into a filtered column, and the synthesis cycle shown in Table 12 was started using a nucleic acid synthesizer (AKTA Oligopilot 10 plus). The desired morpholino monomer compound was added in each coupling cycle to obtain the base sequence of each PMO listed in Table 11 (see Table 12). 【0231】 [Table 12] 【0232】 For the deblocking solution, a dichloromethane solution containing 3% (w / v) trifluoroacetic acid was used. For the neutralization and washing solution, N,N-diisopropylethylamine was dissolved in a 35% (v / v) dichloromethane solution containing acetonitrile, with N,N-diisopropylethylamine at a concentration of 10% (v / v) and tetrahydrofuran at a concentration of 5% (v / v). For coupling solution A, a morpholino monomer compound was dissolved in tetrahydrofuran at a concentration of 0.10 M. For coupling solution B, N,N-diisopropylethylamine was dissolved in acetonitrile, with N,N-diisopropylethylamine at a concentration of 20% (v / v) and tetrahydrofuran at a concentration of 10% (v / v). For the capping solution, a solution of 20% (v / v) acetic anhydride and 30% (v / v) 2,6-lutidine dissolved in acetonitrile was used. The morpholino monomer compounds used were morpholino monomer compound (A), morpholino monomer compound (C), morpholino monomer compound (T), and morpholino monomer compound (G) as listed in Table 13. 【0233】 [Table 13] 【0234】 The aminopolystyrene resin supported with the PMO synthesized above was recovered from the reaction vessel and dried under reduced pressure at room temperature for at least 2 hours. The PMO supported on the dried aminopolystyrene resin was placed in the reaction vessel, 5 mL of 28% aqueous ammonia-ethanol (1 / 4) was added, and the mixture was stirred at 55°C for 15 hours. The aminopolystyrene resin was filtered off and washed with 1 mL of water-ethanol (1 / 4). The resulting filtrate was concentrated under reduced pressure. The resulting residue was dissolved in 10 mL of a mixed solvent (4 / 1) of 20 mM acetate-triethylamine buffer (TEAA buffer) and acetonitrile, and filtered through a membrane filter. The resulting filtrate was purified by reverse-phase HPLC. The conditions used are shown in Table 14. 【0235】 [Table 14] 【0236】 Each fraction was analyzed, the target product was recovered, and concentrated under reduced pressure. 0.5 mL of 2 M phosphoric acid aqueous solution was added to the concentrated residue and stirred for 15 minutes. Then, 2 mL of 2 M sodium hydroxide aqueous solution was added to make it alkaline, and the mixture was filtered through a membrane filter (0.45 μm). The aqueous solution containing the obtained target product was purified using an anion exchange resin column. The conditions used are shown in Table 15. 【0237】 [Table 15] 【0238】 Each fraction was analyzed (HPLC) to obtain the target product as an aqueous solution. The obtained aqueous solution was neutralized by adding 0.1 M phosphate buffer (pH 6.0). Then, desalting was performed by reverse-phase HPLC under the conditions shown in Table 16. 【0239】 [Table 16] 【0240】 The target substance was recovered and concentrated under reduced pressure. The resulting residue was dissolved in water and freeze-dried to obtain each target compound, PMO, as a white, cotton-like solid. The calculated and measured values ​​from ESI-TOF-MS are shown in Table 17 below. 【0241】 [Table 17] TIFF2026095457000049.tif255149TIFF2026095457000050.tif255146TIFF2026095457000051.tif255146TIFF2026095457000052.tif255152 【0242】 Production of antisense PMOs targeting splicing silencers or splice sites involved in single skipping within the introns of the human dystrophin gene. Similar to PMOs that target regions near the donor or acceptor of an intron, the following PMOs have been developed: (i) a PMO having the nucleotide sequence of PMO No. 233 (SEQ ID NO. 257) that targets the splicing silencer sequence or splice site involved in single skipping of intron 44 of the human dystrophin gene; (ii) a PMO having the nucleotide sequences of PMO Nos. 234-244 (SEQ ID NOs. 258-268) that target the splicing silencer sequence or splice site involved in single skipping of intron 45; (iii) a PMO (concatenated type) having the nucleotide sequences of PMO Nos. 245-249 (SEQ ID NOs. 269-273) that target the region near the acceptor of intron 44 (R2) and the splicing silencer sequence or splice site involved in single skipping of intron 45; and (iv) a PMO that targets the splicing silencer sequence of exon 45. PMOs with base sequences No. 250-251 (sequence numbers 274-275) were synthesized. The 5' end of each PMO is a group (3), similar to PMOs that target regions near intron donors or acceptors. Each PMO is shown in Table 18, and the calculated and measured values ​​from ESI-TOF-MS are shown in Table 19. 【0243】 [Table 18] The underlined sequences indicate nucleotide sequences that are expected to correspond to splicing silencer sequences or splice sites involved in single skipping within introns. 【0244】 [Table 19] 【0245】 [Example 2: Multi-exon skipping activity test of antisense oligomers] <Test Example 1> In vitro assay of multi-exon skipping of the human dystrophin gene (1): Induction of multi-exon skipping procedure RD cells (human rhabdomyosarcoma cell line, CCL-136, purchased from ATCC) 3.5 × 10⁻⁶5 For each individual, the antisense oligomers (PMO No. 55, 59, 78, 65, 75, 77, 102, 35, 52, 47, 50, 46, 43, 40, 139, 119, 142, 113, 51, 11, 45, 54, 41, 74, 66, 64, 63, 68, 76, 28, 27, 225, 228, 1, 5, 132, 131, 128, 163, 162, and 161 (Sequence IDs 55, 59, 78, 65, 75, 77, 102, 35, 52, 47, 50, 46, 43, 40, 139, 119, 142, 113, 51, 11, 45, 54, 41, 74, 66, 64, 63, 68, 76, 28, 27, 225, 228, 1, 5, 132, 131, 128, 163, 162, and 161)) were introduced individually or in combinations of 2 or 3 at concentrations of 25-50 μM. The reagents were introduced using the Amaxa Cell Line Nucleofector Kit L with Nucleofector II (Lonza). The program used was T-030. After introduction, the cells were cultured for three nights at 37°C under 5% CO2 conditions in 2 mL of Eagle's minimal essential medium (EMEM) (Sigma-Ace, Inc.) containing 10% fetal bovine serum (FBS) (Invitrogen). After washing the cells once with PBS (Nissui Co., Ltd., the same applies hereafter), 350 μL of Buffer RLT (Qiagen) containing 1% 2-mercaptoethanol (Nacalai Tesque Co., Ltd.) was added to the cells. The cells were left at room temperature for several minutes to lyse, and then collected in a QIAshredder homogenizer (Qiagen). Homogenates were prepared by centrifugation at 15,000 rpm for 2 minutes. Total RNA was extracted according to the protocol provided with the RNeasy Mini Kit (Qiagen). The concentration of the extracted total RNA was measured using NanoDrop ND-1000 (LMS Co., Ltd.). 【0246】 One-step RT-PCR was performed on 800 ng of extracted total RNA using the QIAGEN OneStep RT-PCR Kit (Qiagen). The reaction mixture was prepared according to the protocol provided with the kit. A TaKaRa PCR Thermal Cycler Dice Touch (Takara Bio) was used as the thermal cycler. The RT-PCR program used is as follows: 50°C, 30 minutes: Reverse transcription reaction 95°C for 15 minutes: Polymerase activation, reverse transcriptase inactivation, cDNA thermal denaturation. [94°C, 10 seconds; 57°C, 30 seconds; 72°C, 1 minute] × 33 cycles: PCR amplification 72°C, 10 minutes: Final extension reaction 【0247】 The nucleotide sequences of the forward and reverse primers used in RT-PCR are as follows. Forward primer: 5'-ATTTGACAGATCTGTTGAGAAATGG-3' (SEQ ID NO: 276) Reverse primer 1: 5'-GGCTGTTTTCATCCAGGTTGTG-3' (SEQ ID NO: 277) Reverse primer 2: 5'-AGTTGCTGCTCTTTTCCAGGT-3' (SEQ ID NO: 278) 【0248】 The combination of forward primer and reverse primer 2 can detect transcripts in which multi-exon skipping of exons 45-55 has occurred. Furthermore, the combination of forward primer and reverse primer 1 can detect transcripts in which skipping has not occurred, as well as transcripts in which single exon skipping of exon 45 has occurred. The reaction products of the above PCR were analyzed using Bioanalyzer (Agilent) and MultiNA (Shimadzu Corporation). 【0249】 The polynucleotide amounts "A" of bands in which two or more consecutive exons from exons 45-55 were skipped in numerical order, the polynucleotide amounts "B" of bands in which one exon from exons 45-55 was skipped, and the polynucleotide amounts "C" of bands in which no skipping occurred were measured. Based on these measured values ​​"A", "B", and "C", the skipping efficiency of multi-exon skipping was determined according to the following formula. Skipping efficiency (%) = A / (A + B + C) × 100 【0250】 result The results are shown in Figures 1-16. PMO No. 55, 59, 78, 65, 75, 77, 102, 35, 52, 47, 50, 46, 43, 40, 139, 119, 142, 113, 51, 11, 45, 54, 41, 74, 66, 64, 63, 68, 76, 28, 27, 225, 228, 1, 5, 132, 131, 128, 163, 162, and 161 (Sequence numbers 55, 59, 78, 65, 75, 77, 102, 3 By introducing exons 5, 52, 47, 50, 46, 43, 40, 139, 119, 142, 113, 51, 11, 45, 54, 41, 74, 66, 64, 63, 68, 76, 28, 27, 225, 228, 1, 5, 132, 131, 128, 163, 162, and 161) individually or in combination, multi-exon skipping of exons 45-55 was induced. Therefore, it was found that the antisense oligomers of the present invention described in Table 11, when used alone or in combination, effectively induce or are likely to induce multi-exon skipping. Note that PMO No. 6, 7, 8, 10, 14, 26, 29, 38, 39, 53, 58, 67, 80, 82, 86, 92, 97, 98, 100, 121, 122, 124, 125, 126, 130, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 231, 232 (Sequence numbers 6, 7, 8, 10, 14, 26, 29, 38, 39 It has been confirmed that multi-exon skipping of exons 45-55 can also be induced by introducing (53, 58, 67, 80, 82, 86, 92, 97, 98, 100, 121, 122, 124, 125, 126, 130, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 231, 232). 【0251】 <Test Example 2> In vitro assay of multi-exon skipping of the human dystrophin gene (2): Enhancement of multi-exon skipping procedure RD cells (human rhabdomyosarcoma cell line, CCL-136, purchased from ATCC) 3.5 × 10⁻⁶ 5 For each sample, 25-50 μM of the antisense oligomer (PMO No. 75) from Table 11 was introduced in combination with 25-50 μM of the antisense oligomer (PMO No. 236, 237, or 239) from Table 18, and the assay was performed using the same procedure as in Test Example 1. 【0252】 result The results are shown in Figures 17 and 18. Compared to using PMO No. 75 (SEQ ID NO: 75) alone, the skipping efficiency of multi-exon skipping of exons 45-55 increased when PMO No. 236, 237, or 239 (SEQ ID NO: 260, 261, or 263) was introduced in combination. On the other hand, the skipping efficiency of single-exon skipping of exon 45 decreased. Therefore, it was found that combining the antisense oligomer of the present invention described in Table 11 with the suppressor antisense oligomer of the present invention described in Table 18 promotes, or will promote, multi-exon skipping. 【0253】 <Test Example 3> In vitro assay of human dystrophin gene multi-exon skipping (3): Promotion of overall multi-exon skipping procedure Test Example 3 was performed using the same procedure as Test Example 2, except that the One-Step RT-PCR was performed as follows. One-step RT-PCR was performed on 800 ng of extracted total RNA using the QIAGEN OneStep RT-PCR Kit (Qiagen). The reaction mixture was prepared according to the protocol provided with the kit. A TaKaRa PCR Thermal Cycler Dice Touch (Takara Bio) was used as the thermal cycler. The RT-PCR program used is as follows: 50°C, 30 minutes: Reverse transcription reaction 95°C for 15 minutes: Polymerase activation, reverse transcriptase inactivation, cDNA thermal denaturation. [94°C, 10 seconds; 60°C, 30 seconds; 72°C, 2 minutes] × 40 cycles: PCR amplification 72°C, 10 minutes: Final extension reaction 【0254】 The nucleotide sequences of the forward and reverse primers used in RT-PCR are as follows. Forward Primer 2: 5'-ATTTGACAGATCTGTTGAGAAATGG-3' (SEQ ID NO: 279) Reverse Primer 3: 5'-GGCTCCAATAGTGGTCAGTCC-3' (SEQ ID NO: 280) Forward Primer 3: 5'-CCTGAGAATTGGGAACATGC-3' (SEQ ID NO: 281) Reverse Primer 4: 5'-CCTCCTTCCATGACTCAAGC-3' (SEQ ID NO: 282) 【0255】 Transcripts in which multi-exon skipping has occurred in exons 45-46, 45-47, 45-48, or 45-49 of the human dystrophin gene can be detected using a combination of forward primer 2 and reverse primer 3. On the other hand, transcripts in which multi-exon skipping has occurred in exons 45-46, 45-47, 45-48, 45-49, 45-50, 45-51, or 45-52 can be detected using a combination of forward primer 3 and reverse primer 4. The reaction products of the above PCR were analyzed using Bioanalyzer (Agilent) and MultiNA (Shimadzu Corporation), and the amount of PCR product from each transcript that underwent exon skipping was quantified. 【0256】 result The results are shown in Figures 19-22. Compared to the case of PMO No. 75 (SEQ ID NO: 75) alone, when PMO No. 236 or 239 (SEQ ID NO: 260 or 263) was introduced in combination, the skipping efficiency of multi-exon skipping of exons 45-55 was increased, in addition to the skipping of multi-exon skipping of exons 45-46, exons 45-47, exons 45-48, or exons 45-49, exons 45-50, exons 45-51, and exons 45-52. Therefore, it was found that combining the antisense oligomer of the present invention described in Table 11 with the suppressor antisense oligomer of the present invention described in Table 18 generally promotes, or will promote, multi-exon skipping. [Industrial applicability] 【0257】 The present invention provides antisense oligomers and suppressor antisense oligomers that enable multi-exon skipping of human dystrophin. By using the antisense oligomers and suppressor antisense oligomers of the present invention, novel DMD therapeutic agents and treatment methods are provided.

Claims

[Claim 1] An antisense oligomer, a pharmaceutically acceptable salt thereof, or a hydrate thereof, which simultaneously skips two or more consecutive exons selected from the group consisting of exons 45 to 55 in human dystrophin pre-mRNA, The aforementioned antisense oligomer is The region Rn (where n is an odd number from 1 to 23) of the human dystrophin pre-mRNA, consisting of a sequence of NX bases from the 3' end to the 5' end of the NA-th exon and a sequence of NY bases from the 5' end to the 3' end of the NB-th intron, and The region Rn (where n is an even number from 2 to 24) of the human dystrophin pre-mRNA, consisting of an NX base sequence from the 3' end to the 5' end of the NA intron and an NY base sequence from the 5' end to the 3' end of the NB exon. The sequence includes a nucleotide sequence that is complementary to the nucleotide sequence of at least one region selected from the group consisting of regions R1 to R24, which are shown as R1, R24, and R24, respectively. Here, When n=1, are NA=44, NB=44, NX=20, and NY=400? When n=2, are NA=44, NB=45, NX=600, and NY=50? When n=3, are NA=45, NB=45, NX=20, and NY=400? When n=4, are NA=45, NB=46, NX=400, and NY=50? When n=5, are NA=46, NB=46, NX=20, and NY=400? When n=6, are NA=46, NB=47, NX=400, and NY=50? When n=7, are NA=47, NB=47, NX=20, and NY=400? When n=8, are NA=47, NB=48, NX=400, and NY=50? When n=9, are NA=48, NB=48, NX=20, and NY=400? When n=10, are NA=48, NB=49, NX=400, and NY=50? When n=11, are NA=49, NB=49, NX=20, and NY=400? When n=12, are NA=49, NB=50, NX=400, and NY=50? When n=13, are NA=50, NB=50, NX=20, and NY=400? When n=14, are NA=50, NB=51, NX=400, and NY=50? When n=15, are NA=51, NB=51, NX=20, and NY=400? When n=16, are NA=51, NB=52, NX=400, and NY=50? When n=17, are NA=52, NB=52, NX=20, and NY=400? When n=18, are NA=52, NB=53, NX=400, and NY=50? When n=19, are NA=53, NB=53, NX=20, and NY=400? When n=20, are NA=53, NB=54, NX=400, and NY=50? When n=21, are NA=54, NB=54, NX=20, and NY=400? When n=22, are NA=54, NB=55, NX=400, and NY=50? When n=23, NA=55, NB=55, NX=20, and NY=400, or The antisense oligomer, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein when n = 24, NA = 55, NB = 56, NX = 400, and NY = 50. [Claim 2] Region R1 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 44th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 44th intron. Region R2 is a region consisting of a 600-base sequence from the 3' end to the 5' end of the 44th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 45th exon. Region R3 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 45th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 45th intron. Region R4 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 45th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 46th exon. Region R5 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 46th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 46th intron. Region R6 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 46th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 47th exon. Region R7 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 47th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 47th intron. Region R8 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 47th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 48th exon. Region R9 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 48th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 48th intron. Region R10 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 48th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 49th exon. Region R11 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 49th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 49th intron. Region R12 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 49th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 50th exon. Region R13 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 50th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 50th intron. Region R14 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 50th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 51st exon. Region R15 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 51st exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 51st intron. Region R16 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 51st intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 52nd exon. Region R17 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 52nd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 52nd intron. Region R18 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 52nd intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 53rd exon. Region R19 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 53rd exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 53rd intron. Region R20 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 53rd intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 54th exon. Region R21 is a region consisting of a 20-base sequence in the direction from the 3' end to the 5' end of the 54th exon of the human dystrophin pre-mRNA and a 400-base sequence in the direction from the 5' end to the 3' end of the 54th intron. Region R22 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 54th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 55th exon. Region R23 is a region consisting of a 20-base sequence from the 3' end to the 5' end of the 55th exon of the human dystrophin pre-mRNA and a 400-base sequence from the 5' end to the 3' end of the 55th intron, or The antisense oligomer according to claim 1, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein region R24 is a region consisting of a 400-base sequence from the 3' end to the 5' end of the 55th intron of the human dystrophin pre-mRNA and a 50-base sequence from the 5' end to the 3' end of the 56th exon. [Claim 3] The aforementioned antisense oligomer (a) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, (b) A nucleotide sequence that hybridizes under stringent conditions with a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, (c) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, and having a length within ±15% of the length of the selected nucleotide sequence, or (d) A partial base sequence of any one base sequence selected from the group consisting of (a), (b), and (c) above. An antisense oligomer according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising a complementary base sequence. [Claim 4] The antisense oligomer is an antisense oligomer formed by linking two or more unit oligomers, Each of the aforementioned unit oligomers contains a nucleotide sequence complementary to the nucleotide sequence of any one region selected from the group consisting of regions R1 to R24, or a nucleotide sequence of a part thereof, and the nucleotide sequences of each unit oligomer are not continuous or overlapping with each other. An antisense oligomer according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 5] The antisense oligomer is an antisense oligomer formed by linking two or more unit oligomers, and each of the unit oligomers is (a) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, (b) A nucleotide sequence that hybridizes under stringent conditions with a nucleotide sequence complementary to any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, (c) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 233-256, 341-369, and 385-389, and having a length within ±15% of the length of the selected nucleotide sequence, or (d) an antisense oligomer according to any one of claims 1 to 3, comprising a base sequence complementary to a portion of a base sequence selected from the group consisting of (a), (b), and (c), and wherein the base sequences of each unit oligomer are not continuous or overlapping with each other. [Claim 6] The antisense oligomer according to claim 4 or 5, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein each of the unit oligomers comprises a base sequence complementary to a continuous base sequence of 5 to 20 base lengths in the region. [Claim 7] (1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, or (2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-111, 113-177, 179, 180, 182, 183, 185-193, 195-198, 200-223, and 225-232, and having a length within ±15% of the length of the selected nucleotide sequence. An antisense oligomer according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 8] Sequence numbers 1, 5, 6, 7, 8, 10, 11, 14, 26, 27, 28, 29, 35, 38, 39, 40, 41, 43, 45, 46, 47, 50, 51, 52, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 68, 74, 75, 76, 77, 78, 80, 82, 86, 92, 97, 98, 100, 102, 113, 119, 121, 122, 124, 125, 126, 128, 130, 13 An antisense oligomer according to any one of claims 1 to 3 and 7, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one base sequence selected from the group consisting of 1, 132, 139, 142, 144, 146, 147, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 225, 228, 231, and 232. [Claim 9] An antisense oligomer according to any one of claims 1 to 3 and 7 to 8, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one base sequence selected from the group consisting of SEQ ID NOs: 1, 5, 11, 27, 28, 35, 40, 41, 43, 45, 46, 47, 50, 51, 52, 54, 55, 59, 63, 64, 65, 66, 68, 74, 75, 76, 77, 78, 102, 113, 119, 128, 131, 132, 139, 142, 161, 162, 163, 225, and 228. [Claim 10] An oligonucleotide, an antisense oligomer according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 11] The antisense oligomer according to claim 10, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the sugar moiety and / or phosphate bond moiety of at least one nucleotide constituting the oligonucleotide is modified. [Claim 12] The sugar portion of at least one nucleotide constituting the oligonucleotide has a -OH group at the 2' position that is -OR, -R, -R'OR, -SH, -SR, -NH ２ -NHR, -NR ２ , -N ３ The antisense oligomer according to claim 10 or 11, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, which is ribose substituted with any group selected from the group consisting of -CN, -F, -Cl, -Br, and -I. (The R represents an alkyl or aryl compound, and the R' represents an alkylene compound.) [Claim 13] The antisense oligomer according to any one of claims 10 to 12, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the phosphate bond of at least one nucleotide constituting the oligonucleotide is one selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond, and a boranophosphate bond. [Claim 14] A morpholino oligomer, the antisense oligomer according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 15] The antisense oligomer according to claim 14, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, which is a phosphorodiamidate morpholino oligomer. [Claim 16] The antisense oligomer according to claim 14 or 15, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the 5' end is a group of any of the following chemical formulas (1) to (3). 【Chemistry 15】 [Claim 17] A suppressor antisense oligomer, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, that inhibits single skipping of any one exon selected from the group consisting of exons 45 to 55 of human dystrophin pre-mRNA, The suppressor antisense oligomer is (a) Any one of the nucleotide sequences selected from the group consisting of SEQ ID NOs: 370 to 384, or (b) A nucleotide sequence that has 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of sequence numbers 370 to 384, and has a length within ±15% of the length of the selected nucleotide sequence, or (c) A part of the base sequence of (a) or (b) above Contains a complementary base sequence, The suppressor antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 18] (1) Any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 257 to 275, or (2) A nucleotide sequence having 85% or more sequence identity with any one nucleotide sequence selected from the group consisting of SEQ ID NOs. 257 to 275, and having a length within ±15% of the length of the selected nucleotide sequence. A suppressor antisense oligomer according to claim 17, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising the above. [Claim 19] A suppressor antisense oligomer according to claim 17 or 18, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, comprising any one base sequence selected from the group consisting of SEQ ID NOs: 260, 261, and 263. [Claim 20] An oligonucleotide, the suppressor antisense oligomer according to any one of claims 17 to 19, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 21] The suppressor antisense oligomer according to claim 20, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the sugar moiety and / or phosphate bond moiety of at least one nucleotide constituting the oligonucleotide is modified. [Claim 22] The sugar portion of at least one nucleotide constituting the oligonucleotide has a -OH group at the 2' position that is -OR, -R, -R'OR, -SH, -SR, -NH ２ -NHR, -NR ２ , -N ３ A suppressor antisense oligomer according to claim 20 or 21, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, which is a ribose substituted with any group selected from the group consisting of -CN, -F, -Cl, -Br, and -I. (The aforementioned R represents alkyl or aryl, and the aforementioned R' represents alkylene.) [Claim 23] A suppressor antisense oligomer according to any one of claims 20 to 22, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the phosphate bond of at least one nucleotide constituting the oligonucleotide is one selected from the group consisting of a phosphorothioate bond, a phosphorodithioate bond, an alkylphosphonate bond, a phosphoramidate bond, and a boranophosphate bond. [Claim 24] A suppressor antisense oligomer according to any one of claims 17 to 19, which is a morpholino oligomer, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 25] A suppressor antisense oligomer according to claim 24, which is a phosphorodiamidate morpholino oligomer, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 26] A suppressor antisense oligomer according to claim 24 or 25, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein the 5' end is a group of any of the following chemical formulas (1) to (3). 【Chemistry 16】 [Claim 27] A pharmaceutical composition comprising an antisense oligomer according to any one of claims 1 to 16, a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 28] The pharmaceutical composition according to claim 27, further comprising a suppressor antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, according to any one of claims 17 to 26. [Claim 29] A pharmaceutical composition comprising an antisense oligomer according to any one of claims 1 to 16, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and a suppressor antisense oligomer according to any one of claims 17 to 26, a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 30] (1) The antisense oligomer is an oligomer corresponding to Sequence ID No. 75, and the suppressor antisense oligomer is an oligomer corresponding to Sequence ID No. 260, (2) The antisense oligomer is an oligomer corresponding to SEQ ID NO: 75, and the suppressor antisense oligomer is an oligomer corresponding to SEQ ID NO: 261, or (3) The antisense oligomer is the oligomer corresponding to Sequence ID No. 75, and the suppressor antisense oligomer is the oligomer corresponding to Sequence ID No.

263. The pharmaceutical composition according to claim 28 or 29. [Claim 31] The pharmaceutical composition according to any one of claims 27 to 30, further comprising a pharmaceutically acceptable carrier. [Claim 32] A pharmaceutical composition according to any one of claims 27 to 31 for the treatment of muscular dystrophy. [Claim 33] A pharmaceutical composition according to any one of claims 27 to 32, for administration to a human patient. [Claim 34] A method for treating muscular dystrophy, comprising the step of administering to a patient with muscular dystrophy an antisense oligomer according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, or a pharmaceutical composition according to any one of claims 27 to 33. [Claim 35] The treatment method according to claim 34, wherein the muscular dystrophy patient is a patient who has a mutation in the dystrophin gene that is subject to exon 45-55 skipping. [Claim 36] The treatment method according to claim 34 or 35, wherein the patient is a human being. [Claim 37] Use of an antisense oligomer according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, or a pharmaceutical composition according to any one of claims 27 to 33, in the manufacture of a pharmaceutical for the treatment of muscular dystrophy. [Claim 38] An antisense oligomer according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, for use in the treatment of muscular dystrophy, or a pharmaceutical composition according to any one of claims 27 to 33. [Claim 39] The antisense oligomer, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a pharmaceutical composition according to claim 38, wherein the treatment involves skipping two or more consecutive exons selected from the group consisting of the 45th to the 55th exons of human dystrophin pre-mRNA in numerical order. [Claim 40] The antisense oligomer, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a pharmaceutical composition according to claim 38 or 39, wherein the muscular dystrophy patient is a human being in the aforementioned treatment. [Claim 41] The process includes a step of inhibiting the splicing silencer sequence, splice site sequence, or branch site sequence of the target pre-mRNA when skipping two or more consecutively numbered exons from the target pre-mRNA. A method for increasing the efficiency of skipping two or more consecutively numbered exons. [Claim 42] The method according to claim 41, wherein the splicing silencer sequence is a recognition sequence for heterogeneous nuclear ribbonoprotein A1 (hnRNPA1). [Claim 43] The method according to claim 41 or 42, wherein the target pre-mRNA is human dystrophin pre-mRNA. [Claim 44] The method according to any one of claims 41 to 43, wherein the two or more consecutively numbered exons are selected from the group consisting of the 45th to the 55th exons of human dystrophin pre-mRNA. [Claim 45] The method according to claim 44, wherein skipping of two or more sequentially numbered exons of the target pre-mRNA is performed using an antisense oligomer or a pharmaceutically acceptable salt thereof, or a hydrate thereof, as described in any one of claims 1 to 16. [Claim 46] The method according to claim 44 or 45, wherein the step of specifically inhibiting the splicing silencer sequence, splice site sequence, or branch site sequence is performed using the suppressor antisense oligomer or a pharmaceutically acceptable salt thereof or a hydrate thereof according to any one of claims 17 to 26.

Images