TW200817514A - ENA antisense oligonucleotides exhibiting sequence-specification - Google Patents

Abstract

The present invention provides an antisense oligonucleotides which can specifically cut the sequence to be the target. The oligonucleotides of the present invention comprises an antisense oligonucleotides of the following sequence of formula (I) Wing1-R11-Window-R12-Wing2 (I) [wherein, Window represents a deoxyribouncleotide sequence with 5 or6 nucleotides in number, R11 and R12 maybe the sane or different, each represents a ribonucleotides, Wing1 and Wing2 maybe the same or different, each represents a ribonucleotide, a ribonucleotides sequence, a deoxyribonucleotide, a deoxyribonucleotide sequence or a mixed sequence of a ribonucleotide and a deoxyribonucleotide, but when Wing1 represents a deoxyribonucleotide sequence or a mixed sequence of a ribonucleotide and a deoxyribonucleotide, then in that sequence, the deoxyribonucleotide does not continue more than 4, and when Wing2 represents a deoxyribonucleotide sequence or a mixed sequence of a ribonucleotide and a deoxyribonucleotide, then in that sequence, the deoxyribonucleotide does not continue more than 4, in at least one of ribonucleotide which consisting the sequence of Wing1-R11 or R12-Wing2, the 2'-O and 4'-C of the glycon are bridged by C2-4 alkenyl chain), or the pharmaceutically acceptable salt thereof.

Description

200817514 % ^ IX. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD The present invention relates to a chimeric molecule of an RNA-type modified oligonucleotide having a sequence-specific effect and a DNA oligonucleotide, a composition containing the same, and a composition thereof A method of inhibiting the expression of a target RNA. [Prior Art] Various modified oligonucleotides are designed for antisense or gene evaluation, and several modified oligonucleotides are being subjected to clinical trials [Non-Patent Document 1]. An RNA-type modified oligonucleotide having a Ν-type configuration such as 2·- f '〇Me RNA is known to have high binding affinity to RNA having a complementary sequence. The entire length of the oligonucleotide is applied to translational inhibition, splicing control, etc., as in the case of such an RNA-type modified oligonucleotide [Non-Patent Document 1]. However, the two-stranded strand formed between the RNA-modified oligonucleotide and the mRNA does not serve as a substrate for RNase cleavage of the RNA of the DNA/RNA2 strand, so that the RNA-type modified oligonucleotide is not easily utilized for inhibiting mRNA expression. Antisense law. Therefore, the RNA-type modified oligonucleotide and the DNA oligonucleotide are used as chimeric molecules, and the method of RNase Η activity is adopted.

U used. Such a chimeric oligonucleotide is referred to as a "gapmer", and the DNA is placed in the central portion of the chimeric oligonucleotide called "window", and the RNA-type modified oligonucleotide is placed in a so-called "wing". end. When 2'-〇Me RNA is used as the RNA-type modified oligonucleotide, the gapmer arranged in the wing/window/wing is described as 2'-OMe RNA/DNA/2'-OMe RNA. In order to activate human RNase Η, the DNA portion of the window portion must have a minimum of 4 residues, and the RNase 由 from the coliform must have a minimum of 5 residues [Non-Patent Document 2]. If the chain length of the DNA oligonucleotide in the window portion is long, the 200817514 RNase Η reaction is effective, and if the chain length of the RNA-type modified oligonucleotide of the wing portion is long, the affinity with the target mRNA is improved. . In order to design an appropriate antisense molecule, the appropriate balance between the window portion and the wing portion is such that the DNA oligonucleotide of the window portion is longer than 10 residues, and the high activity as a sense molecule is exhibited [Patent Document 1]. As an RNA-type oligonucleotide, the 2'-oxygen atom of the saccharide moiety is cross-linked with a 4'-carbon atom by an extended ethyl chain (2Ά,4·-(:··ethylidene cross-linked nucleic acid), phase The auxiliary chain nucleic acid exhibits high affinity and has excellent stability to nucleases [Non-Patent Document 3 and Patent Document 2]. There is also vascular endothelial growth factor, and an anti-sense oligonucleotide display for an organic anion transporter. Report of antisense activity in cells [Non-Patent Document 4] As one of the problems of antisense, the antisense oligonucleotide is bound to non-target RNA, and antisense oligonucleotide/non-target RNA 2 strands are cleaved in RNase 、, non-target RNA is cleaved. Chimeric oligonucleotides of DNA oligonucleotides and DNA methylphosphonates are known to avoid such non-specific cuts [Non-Patent Document 5] , 6). The gapmer of 2'-OMe RNA and DNA oligonucleotide cannot be avoided by non-specific cut, or its effect is partial. [Non-Patent Document 7, 8]. Non-Patent Document 1 As shown, the method of lengthening the DNA oligonucleotide of the window portion to 10 or more residues has a concern of non-specific cut. Patent Document 3 and Patent Document 9 describes a design of a gapmer in which a 21-oxygen atom of a head and a 4'-carbon atom are crosslinked by a methylene chain, 2, 4, BNA/LNA and a DNA oligonucleotide. The effect of the gapmer is not mentioned, and the specificity of the cleavage reaction is not mentioned. Patent Document 1: International Publication No. WO2006034348, vol. 200817514 Patent Document 2: Patent No. 3,420,984, Patent Document 3: U.S. Patent Application No. 2006 1 28646 Non-Patent Document 1: Kun*eck, (2003) Eur. J. Biochem. 270, 1628-1644. Non-Patent Document 2: Lima, WF·, Crooke, ST (1 997) Biochemistry 36, 3 90-398 Non-Patent Document 3: Morita K., Hasegawa, C., Kaneko, M., T sutsumi, S., S one, J., Ishikaw a, T., Imanishi, T., Koizumi, M. (2002) Bioorg. Med. Chem. Lett. 1 2, 7 3 -76. Non-Patent Document 4: Koizumi, M. (2006) curr. Opin· Mol. Ther. 8, 144-149· Non-Patent Document 5: Giles, RV And Tidd, DM (1 992) Nucleic Acids Res. 20, 763-770. Non-Patent Document 6: Larrouy, BL Blonski, C., Boiziau, C., Stuer, M., Moreeau, S., Shire D., Tou Lme, J.-J. (1 992) Gene 121, 189-194. Non-Patent Document 7: Larrouy, B., Boiziau, C., Sproat, B.,

Toulme, J.-J. (1 995) Nucleic Acids Res. 23, 3434-3440. Non-Patent Document 8: 31^11, B. 乂., 311 (^11^113, £.11., Eight Games ]^\^1,3· ( 1 99 8) Bioorg Med Chem.6, 1 695 - 1 705. Non-Patent Document 9: Frieden M, Christensen SM, Mikkelsen ND, Rosenbohm C, Thrue CA, Westergaard M, Hansen HF , Orum H, Koch T. (2003) Nucleic Acids Res. 3 1, 636 5 - 6372. 200817514 ^ [Summary of the Invention] The antisense oligonucleotides to be solved by the invention are not only labeled but also bound to non-target RNA. The 2 strands of the antisense oligo/non-target RNA are recognized by RNase , and cleaved by non-standard RNA. Antisense oligos with high specificity are expected to overcome such problems. As a result of reviewing antisense oligonucleotides, the present inventors have found that a gapmer composed of a purine nucleus & gluconate and a DNA oligonucleotide can shorten the DNA oligonucleotide portion of the window. The sequence of the target is specifically cut off, and the present invention has finally been completed. The gist of the present invention is as follows: (1) an antisense oligonucleotide of the following sequence (I) or a pharmacologically acceptable salt thereof,

Wing 1-R11-Window-R12-Wing2 (I) (In sequence (I), Window is a deoxyribonucleotide with nucleotide number 5 or 6, and the nucleotide sequence, R11 and R12 are each a ribonucleotide,

Wing1 and Wing2 are each a ribonucleotide, a ribonucleotide sequence, a deoxyribonucleotide, a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides,

When Wing1 is a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides, the sequence of deoxyribonucleotides in the sequence is not more than 4 consecutive.

Wing2 is a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribose 200817514 nucleotides. In the sequence, there are no more than 4 consecutive deoxyribonucleotides, which constitutes Win^-R11 or R12. - At least one ribonucleotide of the sequence of Wing2 is 2'-0 and 4'-C of the sugar moiety is crosslinked by an alkyl chain). (2) An antisense oligonucleotide according to (1) or a pharmacologically acceptable salt thereof, wherein the Cm alkyl group is an ethylenic chain. (3) An antisense oligonucleotide as described in (1) or (2) or a pharmacologically acceptable salt thereof, which is based on an RNA of a disease-associated gene. (4) The disease-associated gene is an antisense oligonucleotide as described in (3) of the PADI4 gene or a pharmacologically acceptable salt thereof. (5) The target RNA is an antisense oligonucleotide having the gene sequence of the gene library number NM_011061.1 or ΝΜ__0 123 87 as described in (4) or a pharmacologically acceptable salt thereof. (6) A composition comprising the antisense oligonucleotide according to (1) or (2) or a pharmacologically acceptable salt thereof. (7) The composition as described in (6) is used as a medicine. (8) The composition described in (6) is used as a reagent. (9) A method for inhibiting the expression of a target RNA, which is an antisense oligonucleotide or a pharmacologically acceptable salt thereof as described in (1) or (2). (10) The antisense oligonucleotide or the pharmacologically acceptable salt thereof according to (1) or (2), wherein the target RNA is used for the prevention and/or treatment of the disease. (11) The use of the antisense oligonucleotide or the pharmacologically acceptable salt thereof according to (1) or (2) for the manufacture of a medicament for preventing and/or treating a target RNA and a disease. In the present specification, an "antisense oligonucleotide" is an oligonucleotide which can regulate (for example, inhibit, increase -10-200817514) as a target specific gene, and has a target RNA (sense strand) phase. Auxiliary sequence. In the present specification, unless otherwise specified, when the nucleotide chain is represented by an abbreviation, the terminal end is 5, the terminal is left, and the T terminal is right. When the peptide is represented by an abbreviation, the amine terminal is left and the carboxyl terminal is right. EFFECTS OF THE INVENTION The antisense oligonucleotide of the present invention and its pharmacologically acceptable salt are sequence-specific cleavage effects of the target RNA, so that the expression of the target rnA can be specifically inhibited, for example, prevention of the target RNA And / or treatment is effective. The contents of the specification and/or the drawings described in Japanese Patent Application No. 2006-242403, which is the basis of the priority of this case, are included in this manual. [Embodiment] The best shaped bear for carrying out the invention The embodiments of the present invention will be described in detail below. The antisense method using synthetic oligonucleotides is not only used as a pharmaceutical product for diseases such as cancer and viruses, but also for evaluation of genes for pharmaceutical development (J. Kurreck, Eur. J. Biochem, 270, 1 628 (2003).; RS Geary, SP Henry,

L. R Grillone., Clin. Phar mac okinet., 4 1, 255 (2002).; P. Kennewell, Curr. 〇pin. Mol. Ther·, 5, 76 (2003)·). The mechanism of action of the oligonucleotide used in the antisense method is as shown in Fig. 1, and it is a basic concept for binding to RNA in vivo, and it is known that (1) hinders the process of translation of proteins synthesized by mRNA, (2) The process of blocking the splicing of Mrna by the mRNA precursor (pre-mRNA), (3) RNase 反 反 反 反 反 反 反 反 寡 寡 寡 mRNA RN RN 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 The function of mRNA, etc. (J. Kurreck, Eur. J. Biochem., 270,

1 628 (2003).). The general user of the three is the one who uses the RNase H of (3). The antisense oligonucleotide used in this case is a DNA type oligonucleotide. However, when a DNA having a natural phosphodiester bond is used, it is rapidly decomposed by the action of a nuclease in the living body. Therefore, when an antisense method is used in a cell, an oligonucleotide in which a DNA type is modified and a phosphate group is modified as shown in Fig. 2 is used. The special phosphorothioate-modified oligonucleotide (3: PS 0 DN) is nuclease-resistant and forms a two-stranded chain with RNA. It is the substrate of RNase Η and has a certain degree of cell permeability and is most commonly used. (J. Kurreck, Eur. J. Biochem., 270, 1 628 (2003).; RS Geary, SP Henry, L. R Grillone., Clin. Pharmacokinet., 41, 2 5 5(2002).; P. Kenne well, Curr. Opin. Mol. Ther. 5, 76 (2003).). On the other hand, compared with natural DNA, PS ODN has a report that the binding strength of RN A- is reduced, the combination of non-specific proteins, the inhibition of blood coagulation in the living body, and the activity of the complement system are reported. As antisense oligonucleotides that overcome the shortcomings of their PS 0DN, there are many reports of artificial nucleic acids designed using RNA backbones. U. Kurreck, Eur. J. Biochem, 270, 1 628 (2003).; SM Freier, Κ· Η.

Altmann, Nucleic Acids Res., 25, 4429 (1 997).). It is known that the furanose ring constituting the sugar of the nucleic acid mainly forms two kinds of wrinkle patterns as shown in Fig. 2, that is, the N-type configuration and the S-type configuration (W·Saenger, Principles of nucleic acids structure, Springer-Verlag, New York. 1 984.). It is known that the hydroxyl group at the 2' position of the ribose constituting all the nucleosides of the RNA backbone is known as the furanose ring of ribose due to the effect of the -12-200817514 gauche effect of the oxygen atom of the 2'-fluorenyl group and the oxygen atom of the 41 position. The stereo configuration has a large proportion in the N-type configuration. The nucleoside with DNA backbone is the hydrogen atom in the 2' position of ribose, so the gauche effect seen by the RNA backbone is not seen, and the A-shaped helix formed by the natural 2-strand RNA is also preferred in the S-type configuration. Nucleosides exist in the N-type configuration. Actually, the stability is determined by the melting temperature (Tm). The stability of the 2 strands formed by RNA and RN is higher than the stability of the 2 strands formed by DNA and RNA. Report (W. Saenger, Principles of nucleic acids structure, Springe r-Verlag, New York. 1 984.). Because of these, it is advantageous to utilize RNA backbones with the N-type configuration as a priority when considering the use of antisense oligonucleotides targeting RNA. The use of natural RNA as an antisense oligonucleotide is used in cell experiments, and is highly practicable because of the high sensitivity of the nuclease (RNase) present in the serum or cells used for cell culture. Therefore, the RNA osteophyte which is resistant to Rnase is used as a basic derivative. Since the 2^0H group of RNA is required for the decomposition reaction of RNase, most of the derivatives of 2乂〇-alkyl nucleosides (4) which are alkylated by this 2'-0H group and do not become the matrix of RNase are prepared. Reported (Figure 3). Among them, YO-methyl is a modification also seen in tRNA, which is often used in the early days of antisense research (H· Inoue, Y. Hayase, A. Imura, S. Iwai, K. Miura, E. Ohtsuka. Nucleic Acids Res. 1 5, 6 1 3 1 (1 9 87).). The affinity of the 2·-0-methyl group to the complementary strand RNA can also be improved (Δ Tm/mod. (Compared with the case of DNA, it indicates the change of Tm値 at one residue, and the case of + with RNA) High affinity) = + 1.5 ° C ), also showed RNase -13 - 200817514 tolerance. Increasing the carbon number of the alkyl group while reducing the affinity with RNA, in the 2'-〇-propyl group, showing the same ΔTm/mod.値 as the DNA, and above ΔTm/mod.値 (EA Lesnik, C [ Guinosso, AM Kawasaki, Η. S asmor, Μ. Zoune s, LL Cummins, DL Ecker, P. Dan Cook, SM Freier, Biochemistry 32, 78 32 (1 99 3 ).). Highly resistant to nucleases in nucleosides having an alkyl group having 5 or more carbon atoms (EA Lesnik, CJ Guinosso, AM Kawasaki, H. Sasmor, Μ. Zounes, L. B. Cummins, DL Ecker, P. Dan Cook, SM Freier, Biochemistry 32, 7832 (1993).; B., P. Monia, EA Lesnik, C. Gonzalez, WF Lima, D. McGee, CJ Guinosso, AM Kawasaki, P. Dan Cook, SM Freier, J. Biol. Chem. 268, 1 45 1 4 (1 993).; BPMonia, JK Johnson, H. Sasmor, LL Cummins, J. Biol. Chem., 271, 1 453 3 (1 996). . Further, as a substituent at the terminal of the alkyl group, a nucleoside (5: -MOE) having a 2'-fluorenyl-(2-methoxyethyl) group is improved with RNA due to the effect of the methoxyethyl gauche. Affinity (ΔTm/mod. = + 2 °C), also nuclease resistance (P: Martin, Helv. Chim. Acta 7 8, 48 6 (1995).; M. Teplova, G. Minasov, V Tereshko, GB Inamati, P. D an Cook, M. Manoharan, M. Egli, Nature Struct· Biol. 6, 5 3 5 (1 999)·). The nucleoside (6: AP) having 2'-fluorene-aminopropyl group has the same degree of affinity (ΔTm/mod^Ot:) as 2·-〇-propyl group, and the ratio of PS-ODN is observed. Nuclease tolerance (RH Griffey, BP Monia, LL Cummins, S. Freier, MJ Greig, CJ Guinosso, E. Lesnik, SM Manalili, V. Mohan, S. Owens, B. -14- 200817514 R. Ross, H. Sasmor, E. Wancewicz, K. Weiler, PD Wheeler, P. Dan Cook, J. Med. Chem. 39, 5 100 (1996). The reason for this is that the amine group of the 2^0-aminopropyl group of AP is located at the binding site of the metal ion of the 3'-5' exonuclease of the Klenow fragment derived from Escherichia coli, and the nuclease is inhibited by the X-ray crystal structure. Catalytic reaction (M. Teplova, ST Wallace, V. Tereshko, G. Minasov, AM Symons, P. Dan Cook, M. Manoharan, M. Egli, Proc. Natl. Acad. Sci. USA. 96, 14240 ( 1 999)·), an oligonucleotide having a nucleoside (7:2'-F) in which a Y-OH group is substituted with YF is formed by preferentially adopting an N-type configuration, and affinity with RNA is also increased. (△ Tm/mod. = + 2.5〇C) (AM Kawasaki, MD Casper, SM Freier, EA Lesnik, MC Zounes, LL Cummins, C. Gonzalez, P. Dan Cook, J. Med. Chem. 36, 831 ( 1993).) However, it is nuclease-resistant by phosphodiester bond, so it has nuclease resistance as a phosphorothioate bond (AM Kawasaki, MD Casper, S. Μ. Freier, EA Lesnik, MC Zounes, LL Cummins, C. Gonzalez, P. Dan Cook, J. Med. Chem. 36, 83 1 (1 993)·), an artificial nucleic acid completely immobilized to the N-type configuration of RNA, with the sugar department 2: Report of 2W-BNA/LNA (8: cross-linked nucleic acid/locking nucleic acid) in which an oxygen atom and a 4'-carbon atom are crosslinked by a methylene chain (S· Obika, D. Nanbu, Y. Hari, K) Morio,. Y. In, T. Ishi da, T. Imanishi, Tetrahedron Lett., 38, 8735 (1997).; S. Obika, D. Nanbu, Y. Hari, J. Andoh, K. Morio, T Doi, T. Imanishi, Tetrahedron Lett., 39, 540 1 (1 998).; AA Koshkin, SK Singh, P.

Nielsen, V. K. Ra jwanshi, R. Kumar, M. Meldgaard, C. E. -15- 200817514

Olsen, J. Wengel, Tetrahedron, 54, 3 607 (1 998).; S. Obika, T. Uneda, T. Sugimoto, D. Nanbu, T. Minami, T. Doi, T. Imanishi, Bioorg. Med. Chem., 9, 1 00 1 (200 1 )·). Thus, the sugar moiety of the nucleic acid is crosslinked by a methylene chain, and the degree of freedom of the stereo configuration is bound, and the oligonucleotide containing 2, 4'-BNA/LNA exhibits ΔTm/mod. = + 5 - 8 ° C and the stability of the amazing 2 strands. The oligonucleotide containing 2\4'-BNA/LNA also improved the stability of the nuclease as compared with DNA. Further, the 2,4'-BNA/LNA methylene chain is crosslinked with a one-carbon extended ethyl chain (9:2' - 0,4'-C-extended ethyl cross-linked nucleic acid) ΔTm/mod. = + 5°C with the same degree as 2\4'-BNA/LNA, and has better stability against nuclease than 2\4'-BNA/LNA (K. Morita, C. Hasegawa, M. Kaneko, S. Tsutsumi, J. S one, T. Ishikawa, T. Imanisni, M, Koizum, Bioorg. Med. Chem. Lett., 1 2, 73 (2002).; K. Morita, M Takagi, C. Hasegawa, M. Kaneko, S. Tsutsumi, J. Sone, T. Ishikawa, T. Imanish, M. Koizumi, Bioorg. Med. Chem., 1 1 221 1 (2003).). When the saccharide moiety 2, the oxygen atom and the 4'-carbon atom are crosslinked to form a propyl chain, the nuclease resistance is improved, but the decrease is ΔTm/mod. = + 2 °C (K. Morita, M Takagi, C. Hasegawa, M. Kaneko, S. Tsutsumi, J. Sone, T. Ishikawa, T. Imanishi, M. Koizumi, Bioorg. Med. Chem., 11, 22 1 1 (2003).

The RNA type modified antisense oligonucleotide is a 2-strand chain which forms a stable relationship with the target rna as described above. The fully-modified oligonucleotides (hereinafter abbreviated as "FMO"), which are all made of RNA-modified nucleotides, are strongly enhanced in binding ability to mRNA. Therefore, as shown in Fig. 1, (1) inhibition by mRNA synthesis The translation process of proteins, (2) the purpose of blocking the splicing process of mRNA from mRNA precursors is very useful. However, the 2 strands of fm〇 and the target RNA-16-200817514 are not substrates of RNase® (H. Inoue, Y. Hayase, S. Iwai, E. Ohtsuka FEBS Lett. 2 1 5, 327 (1 987).; WF Lima, ST Crooke, Biochemistry 36, 390 (1 997).; H. Wu, WF Lima, ST Crooke, J. Biol. Chem. 274, 28270 (1999). Thus, as shown in Fig. 4A, a chimeric oligonucleotide which is modified by an RNA type nucleoside and DNA is used in an antisense method. It is designed such that the RNA-type modified nucleotide called wing is disposed on both sides of the oligonucleotide, and has a "gapmer" called continuous DNA of the window at the center. That is, the gapmer is a structure having a wing-window-wing. For example, when using 2·-〇-methyl nucleoside (2'-〇Me RNA) as an RNA-type modified nucleoside, it becomes a chimeric oligonucleotide of 2'-〇Me RNA-DNA-2,-〇Me RNA. . Because the gapmer has a continuous DNA field in the central part, the two strands of gapmer and the target RNA become the matrix of RNase. The length of the DNA of this window varies depending on the RNase used. When using 2'-OMe RNA in the wing part, it must be 4 nucleotides or more in human RNase HI (H. Wu, W. F. Lima, ST Crooke, J) Biol. Chem. 274, 28270 (1 999).), above 5 nucleotides in Escherichia coli RNase H (WF Lima, ST Crooke, Biochemistry 36, 390 (1 997).). The longer the DNA field of the window part, the easier it becomes the matrix of RNase. The modified nucleotide of the wing portion is intended to increase the affinity with the target RNA, and the longer the length, the more the affinity with the target RNA is increased. As an antisense activity, the balance between window and wing length is necessary, and it is more active than its best (EA Lesnik, CJ Guinosso, AM Kawasaki, Η. Sasmor, Μ. Zounes, L, L. Cummins, DL Ecker , P. Dan Cook, SM Freier, Biochemistry 32, 7832 (1 993).; BP Monia, EA Lesnik, C. Gonzalez, WF Lima, D. McGee, CJ Guinosso, AM Kawasaki, P. Dan -17- 200817514 • Cook, JSM Freier, J. Biol. Chem. 268, 1 45 1 4 (1 993).; BP Monia, JK Johnson, H. Sasmor, LL Cummins, J. Biol. Chem. 271, 14533 (1996)·) (Fig. 4). 2^〇Me RNA has been used for antisense studies since ancient times, but a recent report on the functional analysis of the PTEN gene by gapmer (M. Sternbergei*, A. Schmiedeknecht, A. Kretschmer, F. Gebhardt, F. Leenders , F. Czauderna, I. von Carlowitz, M. Engle, K. Giese, L. B eigelman, A. Klippel, Antisense Nucleic Acids, Drug Dev. 12,1 3 1 (200 2).). At this time, 2'-〇Me RNA consisting of 7 nucleotides is used in the two wing parts, and PS ODN is made up of 9 nucleotides in the window part, and the antisense is made up of 23 nucleotides in chain length. . The stability of exonuclease is further increased by combining tetrahydrofuran derivatives at both ends. Use the 2'-M〇E for most of the antisense of the gapmer contained in the wing (BA Zinker, CM Rondinone, JM Trevillyan, RJ Gum, JE Clampit, JF Waring, N. Xie, D. Wilcox, P. Jacobson , L. Frost, PE Kroeger, RM Reilly, S. Koterski, ' ; TJ Opgenorth, RG Ulrich, S. Crosby, M. Butler, SF

Murray, RA McKay, S. Bhanot, BP Monia, MR Jirousek, Proc. Natl. Acad. Sci. USA, 99, 11357 (2002).; H. Zhang, J. Cook, J. Nickel, R. Yu, K Stecker, K. Myers, NM De an, Nature Biotechnol. 18, 862 (2000).; BZ Carter, RY Wang, WD Schober, M. Milella, D. Chism, M. Andreeff, Cell Cycle. 2,488 ( 2003)·). 2·-Μ〇Ε For the antisense method, all of the phosphate groups use phosphorothioate linkages. Not only for nuclease tolerance, but also for the adaptation in vivo, the phosphorothioate linkage is anti-sense -18- 200817514 ^ The dynamic surface of the body is more favorable (R. S. Geary, T. A. Watanabe, L.

Truong, S. Freier, E. A. Lesnik, N. B. Sioufi, H. Sasmor, M. Manoharan, A. A. Levin, J. Pharmacol. Exp. Ther. 2 9 6, 890 (200 1).). The 2'-M〇E body formed by the phosphodiester bond is rapidly excreted in the urine, and the 2'-M〇E body formed by the phosphorothioate bond exhibits high blood stability, to the liver, kidney, The migration of the pancreas, bone marrow, etc., beyond the brain. The distribution is better than the PS ODN. The MOE body, which is made up of a phosphorothioate bond, is now clinically developed as an antisense drug against the purpose of anti-tumor, anti-inflammatory, and anti-diabetes. 2,, 4'-BNA/LNA is also used in the same way as 2·-Μ〇Ε, designed as the antisense of gapmer. The antisense of 2W-BNA/LNA containing the total 5 opioid receptor mRNA was designed and injected into the cerebrospinal fluid of mice to observe the inhibition of the pain response of the opioid receptor (C. Wahlestedt, P.

Salmi, L. Good, J. Kela, T. Johnsson, T. Hokfelt, C. Broberger, F. Porreca, J. Lai, K. Ren, M. Ossipov, A. Koshkir i, N. Jakob sen, J. Skouv, H. Oerum, MH Jacobsen, J. Wengel, Proc Natl Acad Sci US A. 97, 5633 (2000).). Furthermore, the FMO antisense of 2',4'-BNA/LNA, which is based on RNA polymerase II, was designed to observe the decrease of RNA polymerase II protein, and the tumor was identified by using the tumor transplantation model in vivo. Proliferation inhibition (K. Fluiter, AL ten Asbroek, Μ. B. de Wissel, Μ. E. Jakobs, M. Wissenbach, H. Olsson, 0. Olsen, H. Oerum, F. Baas, Nucleic Acids Res. 31, 95 3 (2003).). At this time, the phosphate group is not modified, that is, it is used as a phosphodiester bond, and its in vivo dynamics are mainly distributed in the kidney, and it is excreted in the urine. 2, 4^BNA/LNA, which binds strongly to RNA, binds to HIV-1 TAR RNA using a hybrid with 2'-〇Me RNA, showing a function that blocks Tat -19-200817514. This mixture is a complex of a stable chain with a 12-mer, short chain length, a mixture of a polymer and a TAR RNA (A. Arzumanov, AP Walsh, VK Rajwanshi, R. Kumar, J. Wengel, MJ Gait). , Biochemistry 40, 14645 (2001).). In the ENA, the gapmer of the organic anion transporter involved in drug delivery is also shown to be specific to the three subtypes with very similar base sequences (Μ. Takagi, K. Morita, D. Nakai, R. Nakagomi, T. Tokui , M. Koizumi, Biochemistry, 43,4501 (2004)·). The chimeric oligonucleotide made of ENA and 2'-〇Me RNA promotes the atrophy of the gene, and the gene is skipped by the atrophy gene, showing the possibility of treatment (M. Yagi, Y. Takeshima , A. Surono, M. Takagi, M. Koizumi, M. Matsuo, oligo nucleotides, 1 4, 33 (2004).; A. S urono, T. van Khanh, Y. Takeshima, H. Wada, M. Yagi , M. Takag, M. Koizumi, M. Matsuo, Hum. Gene. Ther·, 15, 749 (2004)·). The present invention provides an antisense oligonucleotide having the following sequence (I) or a pharmacologically acceptable salt thereof,

Wingi-RM-Window-R12-Wing2 (I) (In sequence (I), Window is a deoxyribonucleotide sequence of nucleotide number 5 or 6, and R11 and R12 are each a ribonucleotide,

Wing1 and Wing2 are each a ribonucleotide, a ribonucleotide sequence, a deoxyribonucleotide, a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides,

When Wing1 is a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides, the sequence of deoxyribonucleotides in the sequence is not more than four consecutive, -20- 200817514

When Wing2 is a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides, the sequence of deoxyribonucleotides is not more than 4 consecutive, and constitutes Wingl-R11 or R12-Wing2. At least one ribonucleotide of the sequence and at least one ribonucleotide constituting the sequence of R12-Wing2 are each a sugar moiety of 2·-〇 and 4′-C crosslinked by a Cm alkyl chain. The number of bases of the antisense oligonucleotide of the present invention is not limited, and is preferably 8-25, more preferably 10-20, more preferably 12-20. In the sequence (1), the number of nucleotides of Wmg1 and the number of nucleotides of Wing2 are preferably from 0 to 18, preferably from 2 to 13 and from 4 to 3 preferably. In the sequence (I), at least one ribonucleotide constituting the sequence of Wing1-R11 and at least one ribonucleotide constituting the sequence of R 12-Wing 2 are Y-0 and 4'-C of each saccharide moiety. Cm alkyl chain crosslinks. In addition to the ribonucleotides and deoxyribonucleotides constituting the sequence (I), the sugar, the base, the phosphodiester bond, and the terminal phosphoric acid may be modified. Examples of the modification of the sugar may be 2'-oxime-alkylation, 2^0-alkenylation or 2Ά-alkynylation of D-nuclear furanose (for example [Ο-methylation, amine ethylation, 2 , 0-propylation, 2'-0-allylation, 2·-〇-methoxyethylation, 2·-0-butylation, 2′-fluorene-pentylation, 2·-〇 -propargylation, etc.), 2'-oxime of D-nuclear furanose, 4'-oxime alkylation (eg 2, anthracene, 4'-C-extension ethylation, 2, anthracene, 4, C -methyleneation, 2,〇, 4,-C-extension, 2'-〇, 4··C-tetramethylene, etc.), 3'-deoxy-3·-amino group- 2·-deoxy-D-nuclear furanose, 3'-deoxy-3.-amino-2'-deoxy-2 fluorene-D-nuclear furanose, and the like. Examples of the modification of the base may be 5-methylation, 5-fluorination, 5-bromination, 5-iodination, N4-methylation of cytidine, and 5-demethylation of thymidine (uridine). ), 5-fluorination, 5-bromination, 5-iodination, N6-methylation of adenosine, 8-bromination, N2-methylation of uridine, 8-bromo-21-200817514, and the like. Examples of the modification of the phosphodiester bond may be a phosphorothioate bond, a methylphosphonate bond, a methylthiophosphonate bond, a phosphorodithioate bond, an amino phosphate bond or the like. An example of the modification of the terminal phosphoric acid may be esterification of a terminal phosphoric acid or the like. An example of a ribonucleotide in which 2'-0 of a sugar moiety and 4f-C are crosslinked by a Cm alkyl chain are as follows. a compound of the following formula (1) and a salt thereof

[In the formula (1), R1 and R2 are the same or different from each other, a hydrogen atom, a protecting group of a hydroxyl group, a phosphate group, a protected phosphate group or -P(R3)R4 [wherein R3 and R4 are the same or different from each other in a hydroxyl group, Protecting a hydroxyl group, a thiol group, a protected thiol group, an amine group, having a carbon number of 1 to 4 alkoxy groups, a carbon number of 1 to 4 alkylthio groups, a carbon number of 1 to 5 cyanoalkoxy groups or carbon a number of 1 to 4 alkyl-substituted amine groups], A is a C 1 to 4 alkylene group, B is a fluoren-9-yl group, 2-oxo-1,2-dihydropyrimidin-1-yl group, or A substituted inden-9-yl group or a substituted 2-oxo-1,2-dihydropyrimidin-1 -yl group having a substituent selected from the following α group. (α group) · hydroxyl group, protected hydroxyl group, carbon number 1 to 4 alkoxy group, thiol group, protected thiol group, carbon number 1 to 4 alkylthio group, amine group, protected amine group, carbon A number of 1 to 4 alkyl-substituted amine groups, a carbon number of 1 to 4 alkyl groups, and a halogen atom. The "carbon number of 1 to 4 alkylene groups" of the above formula (1) may be, for example, -22-200817514 methylene group, ethylidene group, dimethylene group, tetramethylene group or methylene group. The protecting group of the "protecting group of a hydroxyl group" of R1 and R2 in the above formula (1) and the protecting group of a hydroxy group of R3 and R4 or a group of α may be chemical methods such as hydrogenolysis, hydrolysis, electrolysis, and photodecomposition. A protective group for the cracking of a biological method such as in vivo hydrolysis, such a protecting group may be, for example, a methyl group, an ethyl group, a propyl group, a butyl group, an isobutyl group, a pentamidine group, a pentamidine group, an isovaleryl group, or a hexanyl group. Base, octyl, decyl, decyl, 3-methylindenyl, 8-methylindolyl, 3-ethyloctyl, 3,7-dimethyloctyl, eleventhyl, twelve Sulfhydryl, thirteenth fluorenyl, tetradecyl fluorenyl, fifteen fluorenyl, hexadecanyl, 1-methyl decyl, 14-methyl decyl, 13,13-diyl 14 An alkylcarbonyl group such as anthracenyl group, heptadecyl group, 15-methylhexadecanyl group, octadecyl group, 1-methylheptyl group, 19-mercapto group, 20-mercapto group and 21-mercapto group, a halogenated lower alkylcarbonyl group such as a carboxylic acid, a chloroethylene group, a chloroethane group, a chloroethane group, a trichloroethylene group or a trifluoroethenyl group, such as a dimethyl fluorenyl group, a pentylene group or a hexamethylene group. Low alkoxylower alkane , (Ε)-2-methyl-2-butenyl fluorenyl group, etc., an aliphatic fluorenyl group such as an unsaturated alkylcarbonyl group; an arylcarbonyl group such as a benzinyl group, a naphthylmethyl group or a /3-naphthylmethyl group; a lower alkylated arylcarbonyl group such as a bromobenzylhydrazine group, a 4-chlorobenzyl fluorenyl group, a halogenated carbonyl group, a 2,4,6-trimethylbenzylidene group or a 4-methylbenzyl group, or a 4-methoxybenzyl hydrazide. a carboxylated arylcarbonyl group such as a lower alkoxylated arylcarbonyl group, a 2-carboxybenzyl hydrazino group, a 3-carboxybenzyl fluorenyl group, a 4-carboxybenzyl fluorenyl group, a 4-nitrobenzyl fluorenyl group, a 2-nitrobenzyl hydrazide group "Aromatic fluorenyl group" such as a lower alkoxycarbonylated arylcarbonyl group such as 2-(methoxycarbonyl) fluorenyl group or an arylcarbonyl group such as 4-phenylbenzylidene group; tetrahydrogen; Oral-2,yl-3-bromotetrahydroindole-2-yl, 4-methoxytetrahydroindole-4-yl, tetrahydrothiopyran-2-yl, 4-methoxy Tetrahydrofuran-4-yl or the like tetrahydropyranyl-23-200817514 or tetrahydrothiopyranyl"; tetrahydrofuran-2-yl, tetrahydrofuran-2-yl, etc. "tetrahydrofuranyl or tetrahydrogen Thiofanyl"; trimethyldecyl, triethyldecyl, isopropyldimethylmethyl, tert-butyldimethylalkyl, methyl Tri-lower alkyl fluorenyl group such as isopropyl decyl group, methyl ditributyl decyl alkyl group, triisopropyl decyl alkyl group, diphenylmethyl decyl group, diphenyl butyl decyl group, diphenyl isopropyl group a "decylalkyl group" such as a decyl group, a phenyldiisopropyl decyl group or the like having 1 to 2 aryl groups substituted with a trialkylalkyl group; methoxymethyl, 1,1-dimethyl-1-methoxy "low alkoxymethyl" such as ethoxy group, ethoxymethyl group, isopropoxymethyl group, butoxymethyl group, butoxymethyl group; 2-methoxyethoxymethyl group, etc. Alkoxylated lower alkoxymethyl"; "halo-low alkoxymethyl" such as 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl; 1-ethoxy "Low alkoxyethyl" such as ethyl, 1-(isopropoxy)ethyl; "haloethyl" such as 2,2,2-trichloroethyl; benzyl, α-naphthylmethyl, naphthyl "1 to 3 aryl-substituted methyl groups", such as benzyl, diphenylmethyl, trityl, α-naphthylbenzhydryl, 9-fluorenylmethyl, etc.; 4-methylbenzyl, 2, 4 ,6-trimethylbenzyl, 3,4,5·trimethylbenzyl, 4-methoxybenzyl, 4-methoxyphenyldiphenylmethyl, 4,4^dimethoxytrityl, 2- Nitric acid, 4- Nylbenzyl, 4-chlorobenzyl, 4-bromobenzyl, 4-cyanobenzyl, etc. "The aryl ring is substituted with 1 to 3 lower alkyl, lower alkoxy, halogen, cyano substituted aryl groups. "Methyl"; "low alkoxycarbonyl" such as methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, isobutoxycarbonyl; 2,2,2-trichlorophenoxycarbonyl, 2-trimethyldecylbenzene "low-alkoxycarbonyl group substituted by halogen or tri-lower alkyl fluorenyl group" such as oxycarbonyl; "alkenyloxycarbonyl" such as ethylene oxycarbonyl or aryloxycarbonyl; benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 3, The aryl ring such as 4-dimethoxybenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl or 4-nitrobenzyloxycarbonyl may have 1 to 2 "low-24-200817514 alkoxy or nitro substituted aralkyl" "Oxycarbonyl", "protective group of hydroxyl group" of R1 and R2 is preferably "aliphatic fluorenyl group", "aromatic fluorenyl group", "methyl group which may have 1 to 3 aryl groups substituted", "aryl ring may have 1 to 3 lower alkyl, lower alkoxy, halogen, cyano substituted aryl substituted methyl" or "decyl"" more preferably ethyl, benzinyl, benzyl, methoxybenzidine Base, dimethoxytrityl, monomethyl Trityl or tert-butyl group diphenyl silicon group, R3 and R4 or the α group of "protected hydroxy" should "aliphatic acyl group" or "aromatic acyl group", more suitably acyl benzyl. The protecting group of the "protected phosphate group" of R1 and R2 in the above formula (1) is a protecting group which can be cleaved by a chemical method such as hydrogenolysis, hydrolysis, electrolysis or photolysis or a biological method such as hydrolysis in the human body, and thus protected. The base may be, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, t-butyl, n-pentyl, isopentyl, 2-methylbutyl , neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3- Dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3- "Low alkyl" such as dimethylbutyl or 2-ethylbutyl; "cyanoylated lower alkyl" such as 2-cyanoethyl or 2-cyano-1,1-dimethylethyl; - "Methyl-substituted alkyl", such as methyl diphenyl decyl ethyl, 2-trimethyl decyl ethyl, 2-triphenyl decyl ethyl; 2,2,2-trichloroethyl "halogenated low alkyl" such as 2,2,2-tribromoethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloro-1,1-dimethylethyl; Vinyl, 1-propenyl 2-propenyl, 1-methyl-2-propenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 2-ethyl- 2-propenyl, 1-butenyl, 2-butenyl, benzyl-2-butenyl, 1·25-200817514 methyl-1-butenyl, 3-methyl-2-butenyl, Butyl-2-butenyl, 3-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl, 1-ethyl-3-butenyl, 1-pentyl Candi, 2-pentane, 1·methyl-2-pentyl, 2-methyl-2-pentenyl, 3-pentenyl, 1-methyl-3-pentenyl, 2- Methyl-3-pentenyl, heart pentenyl, 1-methyl-4-pentenyl, 2-methyl-4-pentenyl, hexenyl, 2-hexenyl, 3-hexyl "low alkenyl group" such as alkenyl group, 4-hexenyl group or 5-hexenyl group; cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclobutyl, decyl, adamantyl and the like "Cyanoylated lower alkenyl" such as 2-alkyl; cyanobutenyl; benzyl, α-naphthylmethyl, /3-naphthylmethyl, fluorenylmethyl, phenanthromethyl, fluorenylmethyl, Benzyl, trityl, 1-phenylethyl, 2-phenylethyl, 1-naphthylethyl, 2-naphthylethyl, 1-phenylpropyl, 2-phenylpropyl, 3-phenylpropanoid Base, 1-naphthylpropyl, 2-naphthylpropyl, 3-naphthylpropyl, phenylphenyl, 2-phenylbutyl, 3-phenylbutyl, 4-phenylbutyl, 1-naphthylbutyl, 2 -naphthylbutyl, 3-naphthylbutyl, 4-naphthylbutyl, 1-phenylpentyl, 2-phenylpentyl, 3-phenylpentyl, 4-phenylpentyl, 5-phenylpentyl, bnaphthalene Base, 2-naphthylpentyl, 3-naphthylpentyl, 4-naphthylpentyl, 5-naphthylpentyl, 1-phenylhexyl, 2-phenylhexyl, 3-phenylhexyl, 4-phenylhexyl, 5-phenylhexyl , 6-phenylhexyl, 1-naphthyl, 2-naphthyl, 3-naphthyl, 4-naphthyl, 5-naphthyl, 6-naphthyl, etc. "Aralkyl"; 4-chlorobenzyl, 2- (4-Nitylphenyl)ethyl, o-nitrobenzyl, 4-nitrobenzyl, 2,4-dinitrobenzyl, 4-chloro-2-nitrobenzyl, etc. "The aryl ring has a nitro group and a halogen atom. Substituted aralkyl"; "aryl" such as phenyl, anthracenyl, naphthyl, phenanthryl, anthracenyl; 2-methylphenyl, 2,6-dimethylphenyl, 2-chlorophenyl, 4-chlorobenzene Base, 2,4-dichlorophenyl, 2,5-dichlorophenyl, 2-bromophenyl, 4-nitrophenyl, 4-chloro-2-nitrophenyl, etc. "may have low alkyl, halogen "Atom, nitro substituted aryl", preferably "low alkyl", "cyano substituted lower alkyl", " The aralkyl group or the "aryl-26-200817514 base ring may have a nitro group and a halogen atom-substituted aralkyl group", and more preferably a 2-cyanoethyl group, a 2,2,2-trichloroethyl group or a benzyl group. In the above formula (1), the "carbon number of 1 to 4 oxime groups" of R3 and R4 or the α group is, for example, a methoxy group, an ethoxy group, a n-propoxy group, an isopropoxy group, a n-butoxy group, or a different group. The protecting group of the "protected hydrogenthio group" of R3 and R4 or the α group in the above formula (1) may be a butoxy group, a second butoxy group or a third butoxy group, a methoxy group or an ethoxy group. For example, in addition to the protective group of the above-mentioned hydroxyl group, it may be a "disulfide-forming group" such as an alkylthio group such as a methylthio group, an ethylthio group or a third butylthio group, or an arylthio group such as a benzylthio group. "Alkyl" or "aromatic sulfhydryl" is more suitable for benzidine. The "C1 to 4 alkylthio group" of R3 and R4 or the α group in the above formula (1) may be, for example, a methylthio group, an ethylthio group, a propylthio group, an isopropylthio group, a butylthio group or an isobutyl group. Sulfur, second butylthio, tert-butylthio, methylthio or ethylthio. The protecting group of the "protected amine group" of R3 and R4 or the α group in the above formula (1) may be, for example, a methyl group, an ethyl group, a propyl group, a butyl group, an isobutyl group, a pentyl group or a pentylene group. , isoamyl, hexyl, octyl, decyl, decyl, 3-methylmethyl, 8-methylindenyl, 3-ethyloctyl, 3,7-dimethyloctyl , * mercapto, decyl, thirteenth, thirteenth, fluorenyl, hexadecanyl, hexadecanyl, 1-methyl decyl, 14-methyl decyl, 13, Alkane such as 13-dimethyltetradecyl, heptadecyl, 15-methylhexadecanyl, octadecyl, benzylteenteen, decyl, decyl and decyl a halogenated alkanecarbonyl group such as a carboxyl group, a carbonyl group, a dimethylidene group, a hexamethylene group or a hexamethylene group, a chloroethane group, a dichloroacetinyl group, a trichloroacetic acid group or a trifluoroethylene group; "Alkyl fluorenyl group" such as a lower alkoxy oligocarbonyl -27-200817514 group such as a methoxyethyl group or an unsaturated alkyl carbonyl group such as an (E)-2-methyl-2-butenyl group; Arylcarbonyl such as α-naphthylmethyl, yS-naphthylmethyl, 2-bromobenzyl, 4-chloro a lower alkoxylated arylcarbonyl group such as a haloarylcarbonyl group such as a mercapto group, a lower alkylated arylcarbonyl group such as a 2,4,6-trimethylbenzylidene group or a 4-methylbenzyl group or a 4-methoxybenzyl group; a carboxylated arylcarbonyl group such as a 2-carboxybenzyl fluorenyl group, a 3-carboxybenzyl fluorenyl group, a 4-carboxybenzyl fluorenyl group, a 4-carboxybenzyl fluorenyl group, a 4-nitrobenzyl fluorenyl group or a 2-nitrobenzyl fluorenyl group; "Aromatic fluorenyl group" such as a lower alkoxycarbonylated arylcarbonyl group such as a (methoxycarbonyl)benzylbenzyl group or an arylcarbonyl group such as a 4-phenylbenzylidene group; a methoxycarbonyl group, an ethoxycarbonyl group, and a third butoxy group; a "low alkoxycarbonyl group" such as a carbonyl group or an isobutoxycarbonyl group; a halogen or a tri-low alkyl fluorenyl group-substituted halogenane such as 2,2,2-trichloroethoxycarbonyl or 2-trimethyldecyl phenoxycarbonyl; "Oxocarbonyl"; "alkenyloxycarbonyl" such as vinyloxycarbonyl or aryloxycarbonyl; benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 2-nitrobenzyl The aryl ring such as oxycarbonyl or 4-nitrobenzyloxycarbonyl may have 1 to 2 "low alkoxy or nitro substituted aralkyloxycarbonyl groups", preferably "aliphatic thiol" or "aromatic fluorenyl" ", more suitable benzinyl. In the above formula (1), the "amino group substituted with 1 to 4 alkyl groups" of R3 and R4 or the α group is, for example, a methylamino group, an ethylamino group, a propylamino group, an isopropylamine group, a butylamine group or a different group. Butyryl, second butylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di (second Butyl)amino, di(t-butyl)amino, preferably methylamino, ethylamino, dimethylamino, diethylamino or diisopropylamino. The "C1 to 5 cyanoalkoxy group" of R3 and R4 in the above formula (1) is a group having a cyano group substituted in the above "C1 to 4 alkoxy group", and the group may be, for example, for example, Cyanomethoxy, 2-cyanophenoxy, 3-cyanooxy, 4-cyanobutoxy-28-200817514, 3-cyano-2-mercaptopropoxy, or hydrazine-cyanomethyl-hydrazine , μ dimethyl methoxy, preferably 2-cyanoethoxy. In the above formula (1), the "carbon number of 1 to 4 alkyl groups" of the α group may be, for example, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a second butyl group, or a third group. Butyl, preferably methyl or ethyl. The "halogen atom" of the α group in the above formula (1) may be, for example, a fluorine atom, a chlorine atom, a bromine atom or an iodine atom, a fluorine atom or a chlorine atom. In the above formula (1), all of the appropriate groups of "嘌呤-9-yl" and "substituted -9-yl" are 6-amino -9-yl (also known as adenine D). Amino-protected 6-aminol-9-yl, 2,6-diamine-based -9-yl, 2-amino-6-chloropurha-9-yl, amino group Protected 2-amino-6-chloroindole-9-yl, 2-amino-6-fluorohay-9-yl, amine-protected 2-amino-6-fluoroindole- 9-yl, 2-amino-6-bromoindole-9-yl, amine-protected 2-amino-6-bromoindole-9-yl, 2-amino-6-hydroxyindole-9-yl (also known as guanine), the amine group protected 2_amino-6-hydroxyindole-9-yl, amine group and hydroxyl group protected 2·amino group-6- via quinone-9-yl, 6-Amino-2-methoxyindole-9-yl, 6-amino-2-chloroindol-9-yl, 6-amino-2-fluoroindol-9-yl, 2,6- Dimethoxy 嘌呤9-yl, 2,6-dichloroinden-9-yl or 6-hydrothio-inden-9-yl, more preferably 6-benzylaminomethyl-9-yl, adenyl , 2-Isobutylguanidino-6-hydroxyindole-9-yl or guanine. In the above formula (1), "2-oxy-1,2-a-1" is added to the group - and "substituted 2-oxo-1,2-dihydro-amp; Suitable bases are 2-oxo-4-amino-dihydropyrimidin-1-yl (also known as cytosyl), amine-protected 2-oxo-4-amino-indole, 2-dihydropyrimidine -1 -yl, 2-oxo-4-amino-5-fluoro-1,2-dihydropyrimidin-1-yl, amine-protected 2-oxo-4-amino-5-fluoro-1, 2-dihydropyrimidin-1-yl, 4-amino _ -29- 200817514 2-oxo-5-chloro-1,2-dihydropyrimidin-1-yl, 2-oxo-4-methoxy-1 , 2-dihydropyrimidin-1-yl, 2-oxo-4-hydrothio-1,2-dihydropyrimidine-buyl, 2-oxo-4-hydroxy-1.2-dihydropyrimidin-1-yl (also That is, uracilyl), 2-oxo-4-hydroxy-5-methyl-1, 2-dihydropyrimidin-1-yl (also known as thymidine) or 4-amino-5-methyl-2- Oxy-1.2-dihydropyrimidin-1-yl (also known as 5-methylcytosyl) group, more preferably 2-oxo-4-benzylamino-1,2-dihydropyrimidin-1-yl, Pyrimidinyl, thymidine, uracil, 2-oxo-4-benzylindol-5-methyl-1,2-dihydropyrimidin-1, or 5-methylcytosyl. The antisense oligonucleotide of the present invention is an oligonucleotide-like steroid. "Nucleoside steroid" is a non-natural type of "nucleoside" in which a purine or pyrimidine base is bonded to a sugar. "oligonucleotide steroid" is an unnatural derivative of the same or different "nucleoside" which binds 2 to 50 "oligonucleotides" by a phosphodiester bond, and such a steroid is preferably a sugar moiety. a modified sugar derivative; a thioester derivative in which a phosphodiester bond is partially thiolated; an esterified ester of a terminal phosphoric acid moiety; an amine which is amidated by an amine group on a purine base The saccharide derivative in which the sugar moiety is modified, and the saccharide ester derivative is partially thiolated. The "pharmacologically acceptable salt" is a salt which can be used as a salt by the antisense oligonucleotide of the present invention. Such salts are alkali metal salts such as sodium salt, potassium salt and lithium salt, and alkali metal salts such as salt, magnesium salt, aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts and the like; ammonium salts; And other inorganic salts, third octylamine salt, dibenzylamine salt, morpholine salt, glucosamine salt, phenylglycine alkyl ester salt, ethylenediamine salt, N-methylglucamine salt, barium salt, two Ethylamine salt, triethylamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, chloroprocaine salt, procaine salt, diethanolamine salt, N-benzylphenethylamine An amine salt such as an organic salt such as a salt, a piperidine salt, a tetramethylammonium salt or a tris(hydroxymethyl)aminocarbamate salt; a hydrogenate salt, a hydrochloride salt, a hydrobromide-30-200817514 acid salt, and hydroiodic acid a mineral acid salt such as a salt such as a hydrohalide, a nitrate, a perchlorate, a sulfate or a phosphate; a lower alkanesulfonate such as a methanesulfonate, a trifluoromethanesulfonate or an ethanesulfonate; An acid salt such as an arylsulfonate such as a salt or a p-toluenesulfonate, an acetate, a malate, a fumarate, a succinate, a citrate, a tartrate, an oxalate or a maleate. ; and glycinate, away Amino acid salts such as amine salts, arginine salts, alanine salts, glutamate salts, aspartic acid salts, and the like. A suitable compound of the compound of the formula (1) may be (1) R1 is a hydrogen atom, an aliphatic fluorenyl group, an aromatic fluorenyl group, a methyl group substituted with 1 to 3 aryl groups, a lower alkyl group, a lower alkoxy group. a aryl ring having a halogen or a cyano group substituted with 1 to 3 aryl substituted methyl groups, or a decyl group compound, (2) 111 being a hydrogen atom, an ethyl fluorenyl group, a benzindenyl group, a benzyl group, a p-methoxy group a compound of benzyl, dimethoxytrityl, monomethoxytrityl or tert-butyldiphenylalkyl, (3) R 2 is a hydrogen atom, an aliphatic fluorenyl group, an aromatic fluorenyl group a methyl group, a decyl group or an amino phosphate group having 1 to 3 aryl substituted methyl groups, a lower alkyl group, a lower alkoxy group, an aryl ring having a halogen or a cyano group substituted with 1 to 3 aryl groups. a phosphonium group, a phosphate group or a compound having a protected phosphate group, (4) R 2 is a hydrogen atom, an ethyl fluorenyl group, a fluorenyl group, a benzylidene group, a methoxyheptyl group, a t-butyl diphenyl decyl group. , -P(〇C2H4CN)(N(CH(CH3)2)2), -P(〇CH3)(N(CH(CH3)2)2), phosphonium, or 2-chlorophenyl or 4- a chlorophenyl phosphate group compound, (5) A is a methylene group compound, (6) B is, 6-嘌呤-9-yl (ie adenine), 6-aminopurin-9-yl, 2,6-diaminopurin-9-yl, 2-amino-6-chloro 2-Amino, 6-amino-6-chloroindol-9-yl, 2-amino-6-fluoroindol-9-yl, 2-amino-protected 2-amino-6 -fluoroindole-9-yl, 2-amino-6-bromoindole-9-yl, amine-protected 2-amino-6-bromoindole-9-yl, 2-amino-6-hydroxyindole -9-yl (ie, guanyl-31-200817514), amine-protected 2-amino-6-hydroxyindol-9-yl, amine group and hydroxy-protected 2-amino-6-hydroxyl group嘌呤-9-yl, 6-amino-2-methoxyindole-9-yl, 6-amino-2-chloroindol-9-yl, 6-amino-2-fluoroindol-9-yl, 2,6-dimethoxyfluoren-9-yl, 2,6-dichloropurin-9-yl, 6-hydroxythiofluoren-9-yl, 2-oxo-4-amino-1,2- Dihydropyrimidine-diyl (also known as cytosyl), amine-protected 2-oxo-4-amino-1,2-dihydropyrimidin-1yl, 2-oxo-4-amino-5 -Fluoro-1,2-dihydropyrimidin-1-yl, amino-protected 2-oxo-4-amino-5-fluoro-1,2-dihydropyrimidin-1-yl, 4-amino- 2-oxo-5-chloro-1,2-dihydropyrimidin-1-yl, 2-oxo-4 -Methoxy-1,2-dihydropyridin-1-yl, 2-oxo-4-hydrothio-1,2-hydropyrimidin-1-yl, 2-oxo-4-hydroxy-1 , 2-dihydropyrimidin-1-yl (also known as uracilyl), 2-oxo-4-hydroxy-5-methyl-1,2-dihydropyrimidin-1-yl (also known as thymidine), 4-Amino-5-methyl-2-oxo-1,2-dihydropyrimidin-1-yl (also known as 5-methylcytosyl) group or amine protected 4-amino-5- a compound of methyl-2-oxo-1,2-dihydropyrimidin-1-yl, (7)B is 6-benzylindolyl-9-yl, adenyl, 2-isobutylammonium- 6-Hydroxyindole-9-yl, guanyl, 2-oxo-4-benzylindolyl-1,2-dihydropyrimidin-1-yl, cytosyl, 2-oxo-5-methyl-4 a compound of benzhydryl-1,2-dihydropyrimidin-1-yl, 5-methylcytosyl, uracilidine or thymidine. Further, the above (1) to (2), (3) to (4) or (6) to (7) are more suitable compounds listed as the number is increased. In the formula (1), R1 is from (1) to (2) Arbitrarily selected, R2 is arbitrarily selected from (3) to (4), a is arbitrarily selected by (5), B is arbitrarily selected from (6) to (7), and compounds obtained by any combination thereof are also suitable, and are particularly suitable. A compound selected from the group below. (Compound group) 2, 〇, 4, C-extended ethyl guanosine, ethyl adenosine, 3', 5, di- benzyl benzyl ethyl 6-N-benzyl decyl adenosine, 3 ·, 5·-Di-〇-Benzyl-32- 200817514 • Base-2'-〇, 4'-C-Extended Ethyl-2-N-isobutylguanosine, 5·-〇-Dimethoxytrityl Base 2, hydrazine, 4'-C-extended ethyl-6-indole-benzyl decyl adenosine, 5, fluorenyl-dimethoxytrityl-2f-indole, 4'-C-extended ethyl -2-N-isobutylguanosine, 2-guanidine, 4'-C-extended ethyl-2-1 isobutylguanidinoguanine, 2'-indole, 4, C-extended ethyl-6-N-benzylhydrazine Adenosine, 5'-fluorene-dimethoxytrityl-2, indole, 4, C-extended ethyl-6-N-benzyl decyl adenosine-3^〇-(2-cyanoethyl叱1 diisopropyl)aminophosphoric acid 5'-fluorene-dimethoxytrityl-2'-indole, 4, C-extended ethyl-2-N-isobutylindolino-3.-〇 -(2-cyanoethyl hydrazine, hydrazine-diisopropyl)aminophosphoric acid 2f-indole, 4^C-extended ethyl uridine, 2, hydrazine, 4, C·extended ethyl 5-methyluridine Glycosides, 2, hydrazine, 4, C-extended ethyl cytidine, 2·- ' 〇, 4, C-extended ethyl · 5 · methyl cytidine, di- 〇 - benzyl-2 '- 〇, 4 '-C-Extended ethyl uridine, 5'_〇_dimethoxytrityl-2'- , 4^C-extended ethyl uridine, 3',5, di-indole-benzyl-2'-indole, 4, C-extended ethyl-5-methyluridine, 5·-〇-dimethyl Oxytrityl-2'-indole, 4'-C-extended ethyl-5-methyluridine, di-indolyl-benzyl-2, indole, 4'-C-extended ethyl-4- N-benzyl decyl cytidine, 5'-fluorene-dimethoxytrityl-2, hydrazine, 4, C-extended ethyl-4-N-benzyl cytidine, 3, 5'-two -〇·benzyl-2f-indole, 4, C-extended ethyl-4-N-benzylindol-5-methylcytidine, 5·-〇-dimethoxytrityl-2'- 〇,4,C_Extended ethyl-4-indole-benzylindol-5-methylcytidine, 2·- & 〇,4·-〇 乙基ethyl-4-N-benzyl cytidine, 2·-〇, 4Ά-extended ethyl-4-N-benzylindol-5-methylcytidine, 5'-fluorene-dimethoxytrityl-2, anthracene, 4'-C-stretch Ethyl-uridine-3^0-(2-cyanoethyl N,N-diisopropyl)amino phosphate, 5'-fluorene-dimethoxytrityl-2'-0,4 , (Extended ethyl 5-methyluridine-3L0-(2-cyanoethyl N,N-diisopropyl)amino phosphate, 5·-〇-dimethoxytrityl- 2, 〇, 4'-(:-extended ethyl-4-oxabenzylcytidine-3, 〇-(2-cyanoethyl 1 屮diisopropyl)amino phosphate, and 5'〇- two Oxytrityl-2·-indole, 4·-C-extended ethyl-4-indole benzhydryl-5-methylcytidine-3-0-(2-cyanoethyl hydrazine, hydrazine- Diisopropyl)amino phosphate. -33- 200817514 Specific compounds included in the compound of formula (1) are shown in Tables 1 and 2. But not subject to these restrictions. In Tables 1 and 2, Me is a methyl group, Bn is a benzyl group, Bz is a benzamidine group, PMB is a p-methoxybenzyl group, Tr is a trityl group, and MMTr is a 4-methoxytrityl group ( Monomethoxytrityl), DMTr is 4,4,-dimethoxytrityl (dimethoxytrityl)yl, TMTr is 4,4·,4"-trimethoxy Trityl (trimethoxytrityl) group, TMS is trimethyl decyl, TBDMS is tert-butyl dimethyl decyl, TBDPS is tert-butyl diphenyl decyl, TIPS Is triisopropyldecylalkyl. R3

(V) (Table 1) Illustrative Compound No. A R1 R2 R3 R4 1-1 CH 2 Η Η Η 1-2 1-2 CH 2 Η Η ΜΗ ΜΗ 2 1 -3 CH 2 Η Η Η 〇Η 1 1 4 CH Η Η 〇Η Η 34- 200817514

1 -5 CH 2 Η Η ΟΗ NH 2 1 -6 CH 2 Η Η ΟΗ OH 1 -7 CH 2 Η Η ΝΗ 2 H 1 -8 CH 2 Η Η ΝΗ 2 NH 2 1-9 CH 2 Η Η ΝΗ 2 Cl 1 -10 CH 2 Η Η ΝΗ 2 F 1 -11 CH 2 Η Η ΝΗ 2 Br 1 -12 CH 2 Η Η ΝΗ 2 OH 1 -13 CH 2 Η Η OMe H 1 -14 CH 2 Η Η 〇Me OMe 1 -15 CH 2 Η Η OMe NH 2 1 -16 CH 2 Η Η Cl H 1-17 CH • 2 Η Η Br H 1 -18 CH 2 Η Η F •H 1-19 CH 2 Η Η Cl Cl 1 - 20 CH 2 Η Η SH. H 1 - 21 CH 2 Βη Η NHBz H 1-22 CH 2 Βη· Η OH NHCOCH(CH)2 1-23 CH 2 Βη Βη NHBz H 1 - 24 CH ϊ Βη Βη OH NHCOCHCCH ) 3 2 1-25 CH 2 ΡΜΒ Η NHBz H 1 - 26 CH 2 ΡΜΒ Η OH NHCOCH(CH3)2 1 - 27 CH 2 ΡΜΒ ΡΜΒ NHBz H 1 - 28 CH 2 ΡΜΒ OH OH NHCOCH(CH) 3 2 1 - 29 CH 2 Τγ Η NHBz H 1-30 CH 2 MMTr Η NHBz H 1 -31 CH 2 DMTr Η . NHBz H 1-32 CH Z TMTr Η NHBz H -35- 200817514

1-33 CH 2 Tr Η OH NHCOCH(CH) 3 2 1-34 CH 2 MMTr H OH NHCOCH(CH ) 3 2 1 - 35 CH 2 DMTr H OH NHCOCH ( CH ) 3 2 1-36 CH 2 TMTr H . OH NHCOCH(CH ) 3 2 1 - 37 CH 2 TMS H NHB2 :H 1-38 CH 2 TBDMS ;H NHBz H 1 - 39 CH 2 TBDPS H NHBz H 1 - 40 CH 2 TIPS H NHBz H 1 - 41 CH 2 TMS H OH NHC〇CH(CH ) 3 2 1 - 42 CH 2 TBDMS H OH NHCOCH(CH ) 3 Σ 1-43 CH 2 TBDPS H OH NHCOCH(CH) 3 2 1~44 CH 2 TIPS H :〇H NHC 〇CH(CH ) 3 2 1-45 (CH) 2 2 HHHH 1 - 46 (CH) 2 2 HHH NR 1-47 (CH) 2 2 HHH OH 1 - 48 (CH) 2 2 HH OH H 1-49 (CH) 2 2 HH OH NH 2 1-50 (CH) 2 2 HH OH OH 1-51 (CH) 2 2 HH NH 2 H 1 - 52 (CH) 2 2 HH NH 2 NH 2 1-53 (CH 2 2 HH NH 2 Cl 1 - 54 (CH) 2 2 HH NH 2 F 1 - 55 (CH) 2 2 HH NH 2 Br 1-56 (CH) 2 2 HH NH 2 OH 1 - 57 (CH) 2 2 HH 〇Me H 1 - 58 (CH) 2 2 HH 〇Me OMe 1 - 59 (CH) 2 2 HH OMe NH 2 1-60 (CH) HH Cl H -36- 200817514 1 - 61 (CH) 2 2 Η Η Br Η 1-62 (CH) 2 2 Η Η F Η 1-63 (CH) 2 2 Η Η Cl C1 1-64 (CH) 2 2 Η Η SH Η 1-65 (CH) 2 2· Βη Η Ν Βζ Η 1 - 66 (CH) 2 2 Βη Η 〇Η NHC〇CH(CH) 3 2 1-67 (CH) 2 2 Βη Βη ΝΗΒζ Η 1-68 (CH) 2 2 Βη Βη 〇Η NHC〇CH( CH) 3 2 1-69 (CH) 2 2 ΡΜΒ Η ΝΗΒζ Η 1-70 (CH) 2 2 ΡΜΒ Η 〇Η NHC〇CH(CH) 3 2 1 - 71 (CH) 2 2 ΡΜΒ ΡΜΒ ΝΗΒζ Η 1- 72 (CH) 2 2 ΡΜΒ ΡΜΒ 〇Η NHCOCHCCH) 3 2 1-73 (CH) 2 2 Τγ Η Η Η 1-74 (CH) 2 2 MMTr Η ΝΗΒζ Η 1-75 (CH) 2 2 DMTr Η ΝΗΒζ Η 1 - 76 (CH) 2 2 TMTr Η ΝΗΒζ Η 1-77 (CH) 2 2 Τγ Η 〇Η NHC〇CH(CH) 3 2 1 - 78 (CH) 2 2 MMTr Η OH NHCOCHCCH) 3 2 1 - 79 (CH) 2 2 DMTr Η OH NHCOCHCCH) 3 2 1 - 80 (CH) 2 2 TMTr Η OH NHCOCHCCH) 3 2 1 -81 (CH) 2 2 TMS Η ΝΗΒζ H 1-82 (CH) 2 2 TBDMS Η ΝΗΒζ H 1 - 83 (CH) 2 2 TBDPS Η ΝΗΒζ H 1 - 84 (CH) 2 2 TIPS Η ΝΗΒζ H 1-85 (CH) 2 2 TMS Η OH NHCOCH(CH) 3 2 卜 86 (CH) 2 2 TBDMS Η OH NHCOCH(CH) 3 ; 1-87 (CH) 2 2 TBDPS Η OH NHCOCHCCH) 3 2 1 -88 (CH) TIPS Η OH NHCOCH(CH) -37 200817514 1-89 (CH) 2 3 HH Η H 1 -90 (CH) 2 3 HH H NH 2 1 - 91 (CH) 2 3 HHH OH 1 - 92 (CH) 2 3 HH OH H 1 - 93 (CH) 2 3 HH OH NH 2 1 - 94 (CH) 2 3 HH OH OH 1-95 (CH) 2 3 HH NH 2 H 1 - 96 (CH) 2 3 HH NH 2 NH 2 1 - 97 (CH) 2 3 HH NH 2 Cl 1 -98 (CH) 2 3 HH NH 2 F 1 -99 ( CH) 2 3 HH NH 2 Br 1 -100 (CH) 2 3 HH NH 2 OH 1 -101 (CH) 2 3 HH 〇Me H 1 -102 (CH) 2 3 HH 〇Me OMe 1 -103 (CH) 2 3 HH OMe NH 2 1-104 (CH) 2 3 HH Cl H 1 -105 (CH) 2 3 HH Br H 1 -106 (CH) 2 3 HHFH 1 -107 (CH) 2 3 HH Cl Cl 1 - 108 (CH) ' 2 3 HH SH H 1 -109 (CH ) 2 3 Bn H NHBz H 1-110 (CH) Λ 2 3 Bn H OH nhcoch(ch3)2 1 -111 (CH) 2 3 Bn Bn NHBz H 1-112 (CH) 2 3 Bn Bn OH nhcoch(ch3)2 1-113 (CH) 2 3 PMB H NHBz H 1-114 (CH) 2 3 PMB H OH NHCOCHCCH) 3 2 1 -115 (CH) 2 3 PMB PMB NHBz H 1 -116 (CH) PMB PMB OH NHC〇CH(CH) -38- 200817514

1-117 (CH) 2 3 Tr H NHBz H 1 -118 (CH) 2 3 MMTr H NHBz H 1 -119 (CH) 2 3 DMTr H NHBz H 1 -120 (CH) 2 3 TMTr H NHBz H 1- 121 (CH) 2 3 Tr H OH NHCOCH(CH ) S 2 1 -122 (CH) 2 3 MMTr H OH NHC〇CH(CH ) 3 2 1-123 (CH) 2 3 DMTr H OH NHC〇CH(CH 3 2 1 -124 (CH) 2 3 TMTr H 〇H NHC〇CH(CH ) 3 2 1 -125 (CH) 2 3 TMS H NHBz H 1-126 (CH) 2 3 TBDMS H NHBz H 1 -127 (CH) 2 3 TBDPS H NHBz H 1-128 (CH) 2 3 TIPS H :NHBz H 1 -129 (CH) 2 3 TMS H OH NHCOCH(CI-I)2 1-130 (CH) 2 3 TBDMS H OH NHCOCH(CH) 1-131 (CH) 2 3 TBDPS H OH NHC〇CH(CH ) 3 2 1-132 (CH) 2 3 TIPS H OH NHC〇CH(CH ) 3 2 1 -133 (CH) 2 4 HHHH 1-134 (CH) 2 4 HHH NH 2. 1 -135 (CH) 2 4 HHH OH 1 -136 (CH) * 2 4 HH OH H 1-137 (CH) .2 4 HH OH NH 2 1 -138 (CH) 2 4 HH OH OH 1-139 (CH) 2 4 HH NH 2 H 1 -140 (CH) 2 4 HH NH 2 NH 2 1 -141 (CH) 2 4 HH NH 2 Cl 1-142 (CH) 2 4 HH NH 2 F 1-143 (CH) 2 4 HH NH 2 Br 1-144 (CH ) HH NH OH 2 4 2 -39- 200817514

1-145 (CH) 2 4 Η H 〇Me H 1-146 (CH) 2 4 Η H OMe 〇Me 1 -147 (CH) 2 4 Η H OMe NH 2 1 -148 (CH) 2 4 Η H Cl H 1 -149 (CH) 24 Η H Br H 1 -150 (CH) 2 4· Η HFH 1 -151 (CH) 2 4 Η H Ci Cl 1-152 (CH) 2 4 Η H SH H 1-153 (CH) 2 4 Bn H NHBz H 1-154 (CH) 2 4 Bn H OH NHCOCH(CH) 3 2 1 -155 (CH) 2 4 Bn Bn NHBz H 1-156 (CH) 2 4 Bn Bn OH NHC 〇CH(CH) 3 2 1-157 (CH) 2 4 PMB H NHBz H 1 -158 (CH) 2 4 PMB H OH NHCOCH(CH) 3 2 1-159 (CH) 2 4 PMB PMB NHBz H 1- 160 (CH) 2 4 PMB PMB OH NHCOCH(CH) 32 1-161 (CH) 2 4 Tr H NHBz H 1 -162 (CH) 2 4 MMTr H NHBz H 1-163 (CH) 2 4 DMTr H NHBz H 1-164 (CH) • 2 4 TMTr H NHBz H 1-165 (CH) 2 4 Tr H OH NHC〇CH(CH) 3 2 1 -166 (CH) . 2 4 MMTr H OH NHC〇CH(CH) 3 2 1 -167 (CH) 2 4 DMTr H OH NHCOCH(CH) 3 2 1 -168 (CH) 2 4 TMTr H OH NHCOCH(CH) 3 2 1 -169 (CH) 2 4 TMS H NHBz H 1 - 170 (CH) 2 4 TBDMS H NHBz H 1 -171 (CH) 2 4 TBDPS H NHBz H 1 -172 (CH) TIPS H NHBz H 2 4 -40 200817514 1-173 (CH) Z 4 TMS H OH NHC〇 CH(CH) 3 2 1-174 (CH) 2 4 TBD MS H OH NHCOCH(CH) 3 : 1-175 (CH) 2 4 TBDPS H OH NHCOCH(CH ) 3 2 1 -176 (CH) 2 4 TIPS H OH NHC〇CH(CH ) 3 2 1-177 CH 2 HH OH NHC〇CH(CH ) 3 2 1 -178 CH 2 HH NHBz H 1 -179 (CH) 2 2 HH OH NHCOCH(CH) 3 2 1-180 (CH) 2 2 HH NHBz H 1-181 (CH) 2 3 HH OH NHC〇CH(CH ) 3 2 1 -182 (CH) 2 3 HH NHBz H 1-183 (CH) 2 4 HH OH NHC〇CH(CH) 3 2 1 -184 (CH) HH ; NHBz H 2 4 1-185 CH DMTr P(N(iPr) )(OC H CN) OH NHCOCH(CH) 3 2

1 -186 CH DMTr P(N(iPr) )(OC H CN) NHBz H 1 -187 (CH) 2 2 1 -188 (CH) 2 2 1 -189 (CH) 2 3 1 -190 (CH) 2 3 1-191 (CH) 2 4 1-192 (CH) 2 4 i-193 CH 2 1 -194 CH , 2 1-195 (CH) 2 2 1-196 (CH) 2 2 1-197 (CH) 2 3 1 -198 (CH) 2 3 1-199 (CH) 2 4 1 -200 (CH) 2 4 DMTr P(N(iPr) )(OC H CN) OH NHCOCH(CH )

DMTr P(N(iPr) )(〇C H CN) NHBz H 3 2

DMTr P(N(iPr) )(0C H CN) OH NHCOCH(CH ) 2 2 4 3 2 DMTr P(N(iPr) )(OC H CN) NHBz H

DMTr P(N(iPr)X〇C H CN) OH NHCOCH(CH )

DMTr P(N(iPr)X〇CH CN) NHBz H DMTr P(N(iPr) )(OCH ) OH 2 3 DMTr P{N(iPr) )(OCH) NHBz H 2 3 DMTr P(N(iPr) (OCH) OH 2 3 DMTr P(N(iPr) )(〇CH) NHBz H 2 3 DMTr P(N(iPr) )(OCH ) OH 2 3 DMTr P(N(iPr) )(OCH) NHBz H 2 3 DMTr P(N(iPr) )(〇CH) OH 2 3 DMTr P(N(iPr) )(OCH) NHBz H 2 3 NHCOCH(CH3)2 nhcoch(ch3)2 NHCOCH(CH)2 NHCOCH(CH ) 2 -41 - 200817514 R5

1-201 CH η DMTr P(〇)(〇H)H OH NHCOCH(CM ) 3 2 1 -202 CH 2 DMTr P(0)(0H)H NHBz H 1 -203 (CH) 2 2 DMTr P(0 )(0H)H OH NHCOCH(CH ) 3 2 1 -204 (CH) 2 2 DMTr P(0)(0H)H NHBz H 1 -205 (CH ) 2 3 DMTr P(0)(〇H)H OH NHCOCHCCH ) 3 2 1 -2.06 (CH ) 2 3 DMTr P(0)(0H)H NHBz H 1-207 (CH ) 2 4 DMTr P(〇)(〇H)H OH NHCOCH(CH ) 3 2 1. -208 (CH) DMTr P(0)(0H)H NHBz H 24 (Table 2) i - an example compound

No. A Rl R2 Rr) R6 2-1 CH 2 HH OH H 2-2 CH 2 HH OH CH 2-3 CH HH NH H -42- 200817514 2-4 CH 2 Η Η ΝΗ 2 CH 3 2-5 CH 2 Η Η ΝΗ 2 F 2-6 CH 2 Η Η C1 Η 2-Ί CH 2 Η Η ΗMe Η 2-8 CH 2 Η Η SH Η 2-9 CH 2 Βη Η Η Η 2-10 CH 2 Βπ Βη 〇Η Η 2-11 CH 2 ΡΜΒ Η ΟΗ Η 2 -12 CH 2 ΡΜΒ ΡΜΒ Η Η 2 -13 CH 2 Τ γ Η ΟΗ · Η 2 -14 CH 2 ΜΜΤ γ Η Η Η 2-15 CH 2 DMTr Η * ΟΗ Η 2 -16 CH 2 ΤΜΤγ Η ΟΗ Η 2 -17 CH 2 TMS Η ΟΗ Η 2-18 CH 2 TBDMS Η Η Η 2-19 CH 2 TBDPS Η Η Η 2-20 CH 2 TIPS Η ΟΗ Η 2-21 CH 2 Βη Η ΟΗ CH 3 2-22 CH 2 Βη Βη ΟΗ * CH 3 2-23 CH 2 ΡΜΒ Η ΟΗ CH 3 2-24 CH ΡΜΒ ΟΗ ΟΗ CH 3 2 - 25 CH 2 Τγ Η ΟΗ CH 3 2 - 26 CH 2 ΜΜΤγ Η ΟΗ CH 3 2 - 27 CH 2 DMTr Η ΟΗ CH 3 2-28 CH 2 ΤΜΤγ Η ΟΗ CH 3 2 - 29 CH 2 TMS Η ΟΗ CH 3 2-30 CH 2 TBDMS Η ΟΗ CH 3 2-31 CH 2 TBDPS Η ΟΗ CH 3 -43 200817514

2-32 CH 2 TIPS H OH CH 3 2-33 CH 2 Bn H NHBz H 2-34 CH 2 Bn Bn NHBz H 2-35 CH 2 PMB H NHBz H 2-36 CH 2 PMB PMB NHBz H 2-37 CH 2 Tr H NHBz H 2-38 CH 2 MMTr H NHBz H 2-39 CH 2 DMTr H NHBz H 2-40 CH 2 TMTr H NHBz H 2-41 CH 2 TMS H NHBz H 2-42 CH 2 TBDMS H NHBz H 2-43 CH 2 TBDPS H 丨NHBz H 2-44 CH 2 TIPS H NHBz H 2-45 CH 2 Bn H NHBz CH 3 2-46 CH 2 Bn Bn NHBz CH 3 2-47 CH 2 PMB H NHBz CH 3 2 -48 CH 2 PMB PMB NHBz CH 3 2—49 CH 2 Tr H NHBz CH 3 2-50 CH 2 MMTr H NHBz CH 3 2—51 CH 2 DMTr H NHBz CH 3 2-52 CH 2 TMTr H NHBz CH 3 2 -53 CH , 2 TMS H NHBz CH 3 2-54 CH 2 TBDMS H NHBz CH 3 2-55 CH 2 TBDPS H NHBz CH 3 2 - 56 CH 2 TIPS H NHBz CH 3 2-57 (CH) 2 2 HH OH H 2-58 (CH) 2 2 HH OH CH 3 2-59 (CH) 2 2 HH NH 2 H -44 200817514

2-60 (CH.) 2 2 Η H NH 2 CH 3 2-61 (CH) 2 2 Η H NH 2 F 2-62 (CH) '22 Η H Cl H 2-63 (CH) 2 2 Η H 〇Me H 2-64 (CH) 2 2 Η H SH H 2-65 (CH) 2 2 Bn H OH H 2-66 (CH) 2 2 Bn Bn OH H 2-67 (CH) 2 2 PMB H OH H 2-68 (CH) 2 2 PMB PMB OH H 2-69 (CH) 2 2 Tr H OH H 2-70 (CH) 2 2 MMTr H OH H 2-71 (CH) 2 2 DMTr H ; OH H 2-72 (CH) 2 2 TMTr H OH H 2-73 (CH) 2 2 TMS H OH H 2-74 (CH) 2 2 TBDMS H OH H 2-75 (CH) 2 2 TBDPS H OH H 2- 76 (CH) 2 2 TIPS H OH H 2-77 (CH) 2 2 Bn H OH CH 2-78 (CH) 2 2 Bn Bn OH CH 3 2-79 (CH) . 2 2 PMB H OH CH 3 2 -80 (CH) 2 2 PMB PMB OH CH 3 2-81 (CH ) 2 2 Tr H OH CH 3 2-82 (CH) 2 2 MMTr H OH CH 3 2-83 (CH) 2 2 DMTr H OH CH 3 2-84 (CH) 2 2 TMTr H OH CH 3 2-85 (CH) 2 2 TMS H OH CH 3 2-86 (CH) 2 2 TBDMS H OH CH 3 2-87 (CH) TBDPS H OH CH 2 2 3 -45 200817514

2-88 (CH ) 2 2 TIPS H OH CH 3 2-89 (CH) 2 2 Bn ] H NHBz H 2-90 (CH) 2 2 Bn Bn NHBz H 2-91 (CH) 2 2 PMB H NHBz H 2-92 (CH) 2 2 PMB PMB NHBz H 2-93 (CH) 2 2 Tr H NHBz H 2-94 (CH) 2 2 MMTr H NHBz H 2-95 (CH) 2 2 DMTr H NHBz H 2- 96 (CH) 2 2 TMTr H NHBz H 2-97 (CH) 2 2 TMS H NHBz H 2-98 (CH) 2 2 TBDMS H NHBz H 2-99 (CH) 2 2 TBDPS .H • NHBz H 2- 100 (CH) 2 2 TIPS H NHBz H 2 -101 (CH) 2 2 Bn H NHBz CH 3 2-102 (CH) 2 2 Bn Bn NHBz CH 3 2-103 (CH) 2 2 PMB H NHBz CH 3 2 -104 (CH) 2 2 PMB PMB _z % 2-105 (CH) 2 2 Tr H NHBz CH 3 2-106 (CH) 2 2 MMTr H .NHBz CH 3 2-107 (CH) r 2 2 DMTr H NHBz CH 3 2-108 (CH) 2 2 TMTr H NHBz CH 3 2-109 (CH) „· 2 2 TMS H NHBz CH 3 2-110 (CH) 2 2 TBDMS H NHBz CH 3 2-111 (CH) 2 2 TBDPS H NHBz CH 3 2-112 (CH) 2 2 TIPS H NHBz CH 3 2-113 (CH) 2 3 HH OH H 2-114 (CH) 2 3 HH OH CH 3 2-115 (CH) HH NH H -46 200817514 2-116 (CH) 2 3 Η H NH 2 CH 3 2-117 (CH) 2 3 Η H NH 2 F 2-118 (CH) 2 3 Η H Cl H 2-119 (CH) 2 3 Η H OMe H 2-120 (CH) 2 3 Η HS HH 2 -121 (CH) 2 3 Bn H OH H 2-122 (CH) 2 3 Bn Bn OH H 2-123 (CH) * 2 3 PMB H OH H 2-124 (CH) 2 3 PMB PMB OH H 2-125 (CH) 2 3 Tr H OH · H . 2-126 (CH) 2 3 MMTr H OH H 2-127 (CH) 2 3 DMTr H .OH H 2-128 (CH) 2 3 TMTr H OH H 2-129 (CH) 2 3 TMS H OH H 2-130 (CH) 2 3 TBDMS H OH H 2 -131 (CH) 2 3 TBDPS H OH H 2-132 (CH) 2 3 TIPS H OH H 2 -133 (CH) 2 3 Bn H OH CH 3 2-134 (CH) 2 3 Bn Bn OH CH 3 2-135 (CH) 2 3 PMB H OH CH 3 2-136 (CH ) 2 3 PMB PMB OH CH 3 2-137 (CH) 2 3 Tr H OH CH 3 2-138 (CH> 2 3 MMTr H OH CH 3 2-139 (CH) 2 3 DMTr H OH CH 3 2-140 (CH) 2 3 TMTr H OH CH 3 2-141 (CH) 2 3 TMS H OH CH 3 2-142 (CH) 2 3 TBDMS H OH CH 3 2-143 (CH) 2 3 TBDPS H OH CH 3 -47 200817514

2-144 (CH) 2 3 TIPS H OH CH 3 2-145 (CH) 2 3 Bn H NHBz H 2-146 (CH) 2 3 Bn Bn NHBz H 2-147 (CH) 2 3 PMB H NHBz H 2 -148 (CH) 2 3 PMB PMB NHBz H 2-149 (CH> 2 3 Tr H NHBz H 2-150 (CH) 2 3 MMTr H NHBz H 2 -151 (CH) 2 3 DMTr H NHBz H 2-152 (CH) 2 3 TMTr H NHBz H 2-153 (CH) 2 3 TMS H NHBz H 2-154 (CH) 2 3 TBDMS H NHBz H 2-155 (CH) 2 3 TBDPS H NHBz H 2-156 (CH 2 3 TIPS H NHBz H 2-157 (CH ) 2 3 Bn H NHBz CH 3 2-158 (CH) 2 3 Bn Bn NHBz CH 3 2-159 (CH) 2 3 PMB H NHBz CH 3 2-160 ( CH) 2 3 PMB PMB NHBz CH 3 2 -1:61 (CH) 2 3 Tr H NHBz CH 3 2-162 *(CH) 2 3 MMTr .H NHBz CH 3 2-163 (CH) .· 2 3 DMTr H NHBz CH 3 2-164 (CH) 2 3 TMTr H NHBz CH 3 2-165 (CH) 2 3 TMS H NHBz CH 3 2-166 (CH) 2 3 TBDMS H NHB2 : CH 3 2-167 (CH) 2 3 TBDPS H NHBZ CH3 2-168 (CH) 2 3 TIPS H NHBz CH 3 2-169 (CH) 2 4 HH OH H 2-170 (CH) 2 4 HH OH CH 3 2 -171 (CH) HH NH H -48 200817514 2-172 (CH) 2 4 Η H NH 2 CH 3 2-173 (CH) 2 4 Η H NH 2 F 2-174 (CH) 2 4 Η H . Cl H 2-175 (CH) 2 4 Η H 〇Me H 2-17 6 (CH) 2 4 Η H SH H 2-177 (CH) 2 4 Bn H OH H 2-178 (CH) 2 4 Bn Bn OH H 2-179 (CH) 2 4 PMB H OH H 2-180 ( CH) 2 4 PMB PMB OH H 2 -181 (CH) 2 4 Tr H OH H 2-182 (CH) 2 4 MMTr H OH H 2-183 (CH) 2 4 DMTr H * OH H 2-184 (CH 2 4 TMTr H OH H 2-185 (CH) 2 4 TMS H OH H 2-186 (CH) 2 4 TBDMS H OH H 2-187 (CH) 2 4 TBDPS H OH H 2-188 (CH) 2 4 TIPS H OH H 2-189 (CH) 2 4 Bn H . OH CH 3 2-190 (CH> 2 4 Bn Bn OH CH 3 2-191 (CH) * l 4 PMB H OH CH 3 2-192 ( 2) PFH OH CH 3 2-193 -196 (CH) 2 4 TMTr H OH CH 3 2-197 (CH) 2 4 TMS H OH CH 3 2-198 (CH) 2 4 TBDMS H OH [. CH 3 2-199 (CH) 2 4 TBDPS H OH CH 3 -49 200817514

2-200 (CH) 2 4 TIPS H OH CH 3 2 - 201 (CH) 2 4 Bn H NHBz H 2-202 (CH) 2 4 Bn Bn NHBz H 2-203 (CH) 2 4 PMB H NHBz H 2 -204 (CH) 2 4 PMB PMB NHBz H 2-205 (CH) 2 4 Tr H NHBz H 2-206 (CH) 2 4 MMTr H NHBz H 2-207 (CH) 2 4 DMTr H NHBz H 2-208 (CH) 2 4 TMTr H NHBz H 2-209 (CH) 2 4 TMS H NHBz H 2-210 (CH) 2 4 TBDMS H NHBz H 2 - 211 (CH) 2 4 TBDPS H NHBz H 2-212 (CH 2 4 TIPS H NHBz H 2-213 (CH) 2 4 Bn H NHBz CH 2-214 (CH) 2 4 Bn Bn NHBz CH 3 2-215 (CH) 2 4 PMB H NHBz CH 3 2-216 (CH 2 4 PMB PMB NHBz CH 3 2-217 (CH) 2 4 Tr H NHBz CH 3 2-218 (CH) 2 4 MMTr H NHBz CH 3 2-219 (CH) 2 4 DMTr H NHBz CH 3 2-220 (CH) 2 4 TMTr H NHBz CH 3 2-221 (CH) 2 4 TMS H NHBz CH 3 2-222 (CH) 2 4 TBDMS H NHBz CH 3 2-223 (CH) 2 4 TBDPS H NHBz CH 3 2 -224 (CH) 2 4 TIPS H NHBz CH 3 2-225 CH 2 Η H NHBz H 2-226 CH 2 Η H NHBz CH 3 2-227 (CH) Η H NHBz H 2 2 -50 200817514

2-228 (CH) 2 2 HH NHBz CH 3 2-229 (CH) 2 3 HH NHBz H 2-230 (CH) 2 3 HH NHBz CH 3 2-231 (CH) 2 4 HH NHBz H 2 - 232 ( CH) 2 4 HH NHBz CH 3 2-233 CH 2 DMTr P(N(iPr) )(OC H CN) 2 2 4 OH H 2-234 CH 2 DMTr P(iN(iPr) )(OC H CN) 2 2 4 OH CH 3 2-235 CH 2 DMTr P(N(iPr) )(OC H CN) 2 2 4 NHBz H 2-236 CH 2 DMTr P(N(iPr) )(OC H CN) 2 2 4 NHBz CH 3 2-237 (CH) 2 2 DMTr . P(N(iPr) )(OC H CN) 2 2 4 OH H 2-238 (CH) 2 2 DMTr P(N(iPr) )(OC H CN) 2 2 4 OH CH 3 2-239 (CH) 2 2 DMTr P(N〇Pr) )(〇C;H CN) 2 2 4 NHBz H 2-240 (CH) 2 2 DMTr P(N(iPr) ) (OC H CN) 2 2 4 NHBz CH 3 2-241 (CHA DMTr P(N(iPr) )(OC H CN) 2 2 4 OH H 2-242 (CH2)3 DMTr P(N(iPr) )( OC H CN) 2 2 4 OH CH 3 2-243 (CHA DMTr P(N(iPr) )(OC H CN) 2 2 4 NHBz H 2-244 (CH) 2 3 DMTr P(N(iPr) )( OC H CN) 2 2 4 NHBz CH 3 2-245 (CH) 2 4 DMTr P(N(iPr) )(OC H CN) 2 2 4 OH H 2-246 (CH) 2 4 DMTr P(N(iPr ) ) (OC H CN) 2 2 4 OH CH 3 2-247 (CH) • 2 4 DMTr P(N(iPr) )(OC H CN) 2 2 4 NHBz H 2-248 (CH) 2 4 DMTr P (N(iPr) )(OC H CN) 2 2 4 NHBz CH 2 2-249 CH 2 DMTr P(N(iPr) )(OCH )丨1 3 OH H 2-250 CH 2 DMTr P(N(iPr) )(OCH ) 2 3 OH CH 3 2-251 CH 2 DMTr P(N(iPr) )(OCH ) 2 3 NHBz H 2-252 CH 2 DMTr P(N(iPr) XOCH ) 2 3 NHBz CH 3 2-253 (CH) 2 2 DMTr P(N(iPr) XOCH ) 2 3 OH H 2-254 (CH) 2 2 DMTr P(N(iPr) XOCH ) 2 3 OH CH 3 2-255 (CH) DMTr P(N(iPr) XOCH ) NHBz H 2 2 2 3 -51- 200817514 2-256 (CH) 2 2 DMTr P(N(iPr) )(OCH ) 2 3 NHBz CH 3 2-257 (CH) 2 3 DMTr P(N(iPr) )(OCH ) 2 3 OH H 2- 258 (CH) 2 3 DMTr P(N(iPr) )(OCH ) 2 3 OH CH 3 2-259 (CH) 2 3 DMTr P(N(iPr) )(OCH ) 2 3 NHBz H 2-260 (CH 2 3 DMTr P(N(iPr) )(〇CH ) 2 3 NHBz CH 3 2 - 261 (CH) 2 4 DMTr P(N(iPr) )(OCH ) 2 3 OH H 2-262 (CH) 2 4 DMTr P(N(iPr) )(OCH ) 2 3 OH CH 3 2-263 (CH) 2 4 DMTr P(N(iPr) )(〇CH ) 2 3 NHBz H 2-264 (CH) 2 4 DMTr P(N(iPr) )(OCH ) 2 3 NHBz CH 3 In the above Tables 1 to 2, suitable compounds are (1-5), (1-7), (1-23) (1-24), ( 1-31), (1-35), (1-39), (1 -43), (1-49), (1-51) (1-67), (1-68), (1-75 ), (1-79) , (1 -83), (1-87), (1-93) (1-95), (1-11 1), (1-112), (1-119), (1-123), ( 1- 127), -131), (1-137), (1-139), (1-155), (1-156), (1- 163), (1-167), (1-171) , (1-175), (1-177), (1-178), (1-185), (1-186), (1-193), (1-194), (1-201), ( 1-2〇2), (2-1), (2-2), (2-3), (2-4), (2-10), (2-15), (2-19), ( 2-22), (2-27), (2-31), (2-34), (2·39), (2-43), (2-46), (2-51), (2· 55), (2-57), (2-58), (2-59), (2-60), (2-66), (2-71), (2-75), (2-78) , (2-83), (2-87), (2-90), (2-95), (2-99), (2-102), (2-107), (2-111), ( 2-113), (2-114), (2-115), (2-116), (2-122), (2-127), (2-131), (2-134), (2- 139), (2-143), (2-146), (2-151), (2-155), (2-158), (2-163), (2-167), (2-169) , (2-170), (2-171), (2-172), (2-178), (2-183), (2-187) , (2-190), (2-195), (2-199), (2-202), (2-207), (2- -52- 200817514 211), (2-214), (2- 219), (2-223), (2-225), (2-226), (2-233), (2-234), (2-23 5) or (2·23 6), more suitable for B Guanosine (1-5), 2'-〇, 4|-匕ethyl adenosine (1-7), 3|,5|-di-fluorenyl-benzyl-21-oxime, 4, C- Ethyl-6-N-benzyl decyl adenosine (1-23), 3·,5-di-indole-benzyl-2-indole, 4'-C-extended ethyl-2-indole-isobutyl fluorenyl Guanine (1·24), 5, 〇-dimethoxytrityl-2'-indole, 4'-0-extended ethyl-6-1-benzylbenzyl adenosine (1-31), 5, 〇-dimethoxytrityl- 2·-KC-extended ethyl-2-N-isobutylguanosine guanosine (1-35), 2, 〇, 4Ά-extended ethyl-2-N- Isobutyl guanosine (1-177), 2'-〇, 4, C-extended ethyl-6-1-benzyl decyl adenosine (1-178), 5^〇-dimethoxytrityl- 2, 〇, 4'-0-extended ethyl-2-indoleisoguanosine guanosine-3, 〇-(2-cyanoethyl 1 屮 diisopropyl) amino phosphate (1-185), 5 two Methoxytrityl-oxime:-extended ethyl-6-N-benzyl decyl adenosine 0-(2-cyanoethyl N,N-diisopropyl)amine Phosphate (1-186), 2'-〇, 4, C-extension ethyl uridine (2-1), 2, 〇, 4-C-extended ethyl 5-methyluridine (2-2) , 2, 〇, 4'-C-extended ethyl cytidine (2-3), 2'-〇, 4^-extended ethyl-5-methylcytidine (2-4), 3', 5' -Bis-benzyl-benzyl-2'-indole, 4·-c-extended ethyl uridine (2-1〇), 5Ά-dimethoxytrityl me-extended ethyl uridine (2- 15), 3',5--di-indole-benzyl-2, indole, 4, C-extended ethyl-5-methyluridine (2-22), 5, fluorene-dimethoxytriphenyl Methyl-2, indole, 4'-C-extended ethyl-5-methyluridine (2-27), 3·,5·-di-0-benzyl-r-0,4'-C- Ethyl 4-N-benzyl cytidine (2-34), 5'-fluorenyl-dimethoxytrityl-2·-indole, 4, (:-extended ethyl-4-indole -benzylbenzyl cytidine (2-39), 3\5·-di-indole-3-benzyl-2, anthracene, 4, C-extended ethyl 4-n-benzylindol-5-methylcytidine (2-46), 5'-〇-dimethoxytrityl-2|-indole, 4^0-extended ethyl-4-indolebenzyl-5-methylcytidine (2-51 , 2, 〇, 4^-Extended ethyl-4-N-benzyl cytidine (2-225), 2'-〇, 4'-C-extended ethyl-4-N-benzyl hydrazine-53 - 200817514 - keto-5-methylcytidine (2-226), 5, 〇-dimethoxy Triphenylmethyl-2_-indole, 4^-ethyl-uridine-3'-indole-(2-cyanoethyl N,N-diisopropyl)amino phosphate (2-233), 5'-〇--*methoxydimethyl--2, oxime, 4, C-extended ethyl _5-methyluridine-3·-0-(2-cyanoethyl N,N-di Isopropyl)amino phosphate (2-234), 5·-〇-dimethoxytrityl K, 4, C-extended ethyl-4-N-benzyl cytidine- 3·- 〇-(2-cyanoethyl N,N-diisopropyl)amino phosphate (2-235), or, 5·-〇-dimethoxytrityl 2, 〇, 4·- (:-Extended ethyl-4·Ν-benzylindol-5-methylcytidine-3'-indole-(2-cyanoethylguanidine, fluorene-diisopropyl)amino phosphate (2-23 6). ί $ The compound of the formula (1) can be produced by the method of JP-A-2000-297097. The antisense oligonucleotide of the present invention and a pharmacologically acceptable salt thereof may also be present as a solvate (hydrated), and thus a solvate is also included in the present invention. The RNA to which the antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof is the target is not limited, and for example, RNA of a gene involved in the disease is preferred. The disease can be exemplified as follows. 1. Viral disease Respiratory syncytial virus, giant cells. Virus, hepatitis C virus, hepatitis B virus, herpes simplex virus, papillomavirus, Epstein-Barr virus, influenza virus, lime mosaic virus, Sinai virus, HIV, etc. 2. Inflammatory disease Crohn's disease, ulcerative large intestine Inflammation, chronic joint rheumatism, asthma, dryness, atopic dermatitis 3. Metabolic syndrome such as metabolic related diseases, diabetes, obesity, hyperlipidemia, hypercholesterolemia-54-200817514, hypertriglyceridemia, etc. 4 Cardiovascular disease, familial hypercholesterolemia, non-familial hypercholesterolemia, lipemia, /3 lipoproteinemia, atherosclerosis, coronary artery disease, myocardial infarction, hypertension, carotid artery disease , stroke, peripheral vascular disease, thrombosis or arterial arterial pain, etc. 5 · cancer skin, connective tissue, fat, chest, lung, stomach, pancreas, ovary, neck, uterus, kidney, bladder, large intestine, prostate, Central nervous system (CNS), retinal or circulatory cancer (such as leukemia and lymphoma) 6. Central diseases, Alzheimer's disease, Parkinson's disease, muscle atrophic lateral sclerosis (ALS), etc. As for the genes related to the disease, it can be exemplified by respiratory syncytial virus, cytomegalovirus, hepatitis C virus, hepatitis B virus, herpes simplex virus, papilloma virus, Epstein-Barr virus. , influenza virus, lime mosaic virus, Sinai virus, or HIV gene, PADI4, PTEN, tumor necrosis factor receptor-associated death region (TRADD), glucocorticoid receptor (GCCR), dimercaptoglycerol thiol transferase 2 (DGAT2), ApoB-100, ICAM-1, protein tyrosine phosphatase IB (PTPIB), interleukin 4 receptor (IL4R-a), C-reactive protein (CRP), glucagon (GCGR), VLA_4 (final antigen_4), Clusterin, insulin-like growth factor-I receptor (IGF-1R), survivin, eukaryotic initiation factor-4B (eIF-4E), c-Raf kinase , heat shock protein 27 (Hsp27), Cu/Zn superoxide dismutase (S0D1), telomerase, -55- 200817514 bcl-2, VEGF, VEGF-R, Hif-1 α, H-Ras, N -Ras, K-Ras, TNF_R, ribonucleotide reductase (RNR), and the like. The antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof may also be a nucleic acid molecule encoded by, for example, peptidyl arginine deiminase 4 (hereinafter referred to as PADI4 enzyme) (for example, mature mRNA, mature mRNA precursor, template) DNA, etc.) are the subject. One of the base sequences of the nucleic acid molecule encoded by the PADI4 enzyme is shown, for example, in SEQ ID NOs: 1 and 3. Sequence numbers 1 and 3 each represent the base sequence of mouse and human PADI4 mRNA. The nucleic acid molecule encoded by the PADI4 enzyme may also be a nucleic acid molecule encoded by a protein which hybridizes under stringent conditions to a nucleic acid molecule having a sequence of SEQ ID NO: 1 or 3 and which has a biological activity of PADI4 enzyme. The biological activity of the PADI4 enzyme is to act as an antigen in addition to the activity of the catalyst for the deamidation of the arginine residue in the protein to a citrulline residue in the presence of calcium ions. As the activity of the immunogen. Further, one of the amino acid sequences of the PADI4 enzyme is shown, for example, in SEQ ID NOs: 2 and 4. SEQ ID Nos. 2 and 4 are amino acid sequences encoded by the nucleotide sequences of each of Sequence Nos. 1 and 3. The amino acid sequence of SEQ ID NO: 2 or 4 is obtained by deleting, substituting or adding an amino acid sequence of one or several amino acids, and a protein having a biological activity of PADI4 enzyme is also included in the PADI4 enzyme. The antisense sequence is usually a functional site selected from a plurality of mRNAs, such as a 5'-non-translation site, a start codon, a splicing site, a stop codon, and the like. There are also several methods for determining antisense sequences based on the nucleotide sequence of the target gene. For example, a method of mixing a random short-stranded nucleic acid fragment with an RNA of a target gene, digesting with RNase Η, and using a cleavage site as an antisense sequence (refer to -56-200817514, for example, Lloyd, Β·Η· et al, Nucleic Acids) Research, 2001, 29, P3 664-3673. Ho, SP et al., Nucleic Acids Research, 1996, 24, pl 901 - 1 907. Matveeva, 0·, 1 997, 25, p50 10-501 6.); Mixing a random short-stranded DNA fragment with the rna of the target gene, and selecting the binding site by a gel shift method or a recovery method using an oligo dT vector, and determining the binding site (see, for example, Bruce, TW and L ima, WF) , Biochemistry, 1 997, 36, p5004-501 9. Takagi, M. et al

Biochemistry, 2004, 43, 450 1)), an oligonucleotide having an antisense sequence of 20 to the extent of immobilization on a glass plate or the like to prepare a matrix for hybridization with RNA of a target gene identified by a radioisotope or the like, A method for determining the sequence of RNA binding (see, for example, Sohail, M. et al., Nucleic Acids Research, 200 1, 29, ρ2041 - 2051·) and the like. There are also reports of methods for determining antisense sequences using computer programs (see, for example, Scherr, Μ., Nuc 1 eic Acids Research, 2000, 28, p2455-2461. Patzel et al, Nucleic Acids Research, 1 999, 27, p4328 -4334.). In this way, the complex high-order structure of mRNA can be used to find out the part of the strand domain in which the antisense molecule is easily bound. The base sequence of the antisense oligonucleotide of the present invention which is based on PADI4 mRNA is shown, for example, in SEQ ID NOs: 3 and 4. The nucleotide sequences of SEQ ID Nos. 3 and 4 are sequences complementary to nucleotide numbers 564-581 and 867-887 of each gene library number NM_01106 1.1. The antisense oligonucleotide of the present invention which is based on PADI4 mRNA is suitable as exemplified below. (1) HOTe2-ΟΤβ2-T-5Ce2-G-Te2-G-OT-TA-Ge2-G-Ge2-T-5Ce2-Ae2t-H (AS-1-2 57- 200817514 (2) HO-Te2-5Ce2- T-Te2-C_Ge2-T_G-CTTA-Ge2-G-Ge2-T-5Ce2~Ae2t-H (AS-Bu 1) (3) H〇-Ae2-C-Ge2-TC-Ae2-C-.A62-CTGTC- Te2-T-Ge2-GA-Ae2-CA must-H (A S-2-3)

(4) H〇-Te2-C-Te2-T-5Ce2_G-T(GCTTA-Ge2-C"Ge2-T-5Ce2t-H (5) HO-Te2-C-Te2-丁-5Ce2-G-Te2- GCTTAG°2~G-Ge2-f -H (6) H〇-T32-OTe2-丁-5Ce2-G-Te2—G—OT-TA-Ga2-G-Ge2t-H (7) HO-

(8) H〇-T'°2-T-5Cf2-G-Te2·-GC-TTA-Ge2-G-Ge2—T-5Ce2-A must-H (9) HOT-5C°2-G-T32-G-OT -TA-Ge2-C Guang Ge2-T-5Cf2-/V2t-H

(10) H〇-5C°2-G-Te2-G-OT-T-A-C, 2-G-G^T-5Ce2-As2t-H

(11) H〇-C-Te2-T-5Ce2-G-Te2-G-C-T-_T-A-Ge2-G-Ge2-T-5Ce2T•一H

(12) HO-Te2-T-5Ce2-G-Te2-G-C-T-T-A-G^-G-G dip-f-H

(13) H〇_T~5Ce2-G-Te2-G-C-T-T-A-Ge2-G-G must-H

(13) H〇-5C"2-G-Ta2-G»C~T-T-A-Ge2-G-Gfi2t-~H

(14) H〇-TG2-5Cg2-T-Te2-OGe2-T-G-C-T-T-A-G02-G-Ge2-T-5C, 21-H

(15) HO-Te2-5C(T-Te2-C-Ge2-T-G-C-T-T-A-G°2-G_Ge2-Tl-H

(16) H〇-T'2-5Ce2-T-Te2-C-G°2-T-G-OT-T-A-Ge2-G-Ge2t-H

(17) HO—5Ce2-T-Te2-C-Ge2-T-G—C-T-T-A—Gh2-G-Ge2-丁-5Ce2-Ae21—H

(18) H〇-T-Te2-OC/2-T-G-OT-T-A-Ge2-G-Ge2-T-5Ce2—A must-H

(19) H〇-Te2-OGe2-T-G-OT-T-A-Ge2-CrGe2-T-5Ce2-Ae2t-H

(20) H〇-5CG2-T-Te2-C*-G〇2--T--G--C--T--T-A~Cxe2--G-Ge2-T-5Ce2l~H

(21) H〇-T-Te2-C-Ge2—T-G—C-T-T-A-Ge2-G-Ge2-f-H

(21) HO-Te2-C-Ge2-TGCTTA-Ge2-G-Ge2-H (22) H〇—A02—C—Ge2-T—C—Ae2-OAa2-OT-GT-C, Te2-butyl- Ge2-GA-A02-CH-1

(23) H 0-Ae2-OGe2-T-C-Ae2-C-Aa2-C-T-G-T-C-T°2-T-Ge2-G-A-Ae2i-H

(24) H〇—Ae2-C~Ge2-T-C-Ae2-OAs2—C-T-G-T-OTe2-T-Ge2-CrAl-H-58- 200817514

- (25)H〇-Ae-C-Ge2-T~C-Ae2~C-Ae2-CTGT~C-Te2~T-Ge2-Gt~H (26) HOAe2-OGe2-TC-Ae2-C-Ae2 -CT-GTC-Te2-T-Ge2t-Η

(27) H〇-OGe2-T-C-Ae2-〇Ae2-OT-G-T-〇Te2-T-Ge2-G-A-Ae2-C-Ae2t-H

(28) H〇-Ge2—T-〇Ae2—C—Ae2-〇TGT-〇Te2—T-Ge2-GA-Ae2-〇Ae2t-H (29) H〇-TC-Ae2-C-Ae2-ΟT -GTC-Te2-T-Ge2-GA-Ae2-〇Ae2t-Η

(30) HOOAe2-〇Ae2-CTGT-〇Te2-T-Ge2-GA-Ae2-C-Ae2i-H (31>H〇-Ae2-〇Ae2-ΟΤ-GT-ΟΤ02-T-Ge2-GA-Ae2 -C-Ae2t~H

(32) H〇~OG32-T-C-Ae2- OA°2 - C—T a G-T-〇Te2-T - Ge2-G-A- Ae2 - C^H

(33) HOGe2-T-C-Ae2-C-Ae2-C-T-G-T-C-Te2-T-Ge2-G-A-Ae2t-H

(34) HOT-OAe2-OAe2-CTG-丁-OTe2-T-Ge2-GA^-H (35) H〇-C-Ae2-C-Ae2 - CTGTC-r2 - T-G"2-σ-Αι -Η (36) HO-Ae2- OAe2-CT - GT -

(37) HOAe2-〇Ae2-OT-G~T-〇Te2-T-Ge2t-H In this manual, A, G, C, 5C, T, A\ G\ C\ 5C\ As, Gs, (T, 5CS, Ts, Ae2, Ge2, 5Ce2, Te2, Ae2t, Ge2t, 5Ce2t, Ae2s, Ge2s, 5Ce2s, Te2s, Ael, Gel, Gelt, 5Ce1, Tel, Aels, Gels, 5C' and Teis are each of the following formula (8), (G), (C), (5C), (T), (A\ (G), (Cl (5C1 (Tl (As), (Gs)n {C\ (5Cs), (Ts)n (Ae2 )N (Ge2)N (5Ce2), (Te2)N (Ae2t)N (Ge2t) N (5Ce2\ (Ae2s)N (Ge2s), (5Ce2s), (Te2s), (Ael), (Gel)N ( Gel\ (5Cel), (Τθ1), (Aels), (Gels), (5Cels) and (Teis), -59- 200817514

-60- 200817514[化5]

°χ

St〇H °N ο

(A"2)

(Ge2) (A·28) 0

(G must)

(5Ce2) (SC4»28)

0

o

(r2) (r2s) -61 - 200817514 [Chem. 6]

(A«1)

S-P-OH (Ae1s) Ο

Ο

〇=Ρ-〇Η Ο

(G·1” (Get)

(5Cd1) (5C·1" Ο

Ο

S=P~OH (r1) (r1s) -62- 200817514 The antisense oligonucleotide of the present invention and the pharmacologically acceptable salt thereof have high binding affinity to RNA, high tolerance to nucleases, and can be caused by RNase The decomposition of mRN A can be specifically accepted in sequence. Therefore, the antisense oligonucleotide of the present invention and its pharmacologically acceptable salt can inhibit the expression of the target RNA. The antisense oligonucleotide of the present invention and its pharmacologically acceptable salt are effective for the treatment and/or prevention of the target RNA and the disease. The antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof can be used to produce a medicament for preventing and/or treating a target RNA and a disease. The antisense oligonucleotide of the present invention and its pharmacologically acceptable salt can be used for medicine or a drug. The antisense oligonucleotide of the present invention and its pharmacologically acceptable salt can be used for the treatment and/or prevention of a target RNA for diseases and diseases. Further, the antisense oligonucleotide of the present invention and its pharmacologically acceptable salt can be used in a test tube, in vivo or ex vivo. The antisense oligonucleotide of the present invention can be used in a commercially available synthesizer (e.g., model 392 of the Amino Phosphate Method of Barking Emma), and the like (Nucleic Acids Research, 12, 4539 (1 984)). Method synthesis. The amino phosphate reagent used at this time is a natural type of nucleoside and Y-0-methyl nucleoside (ie, 2'-〇-methylguanosine, 2'-〇-methyladenosine, 2Ά-A A cytosine, 2'-indole-methyl uridine, a commercially available reagent can be used. The carbon number of the alkyl group is 2-6 2^0-alkylguanosine, adenosine, cytidine and uridine as follows. 2·-〇-aminoethylguanosine, adenosine, cytidine, uridine can be synthesized according to the literature (Blommers et al Biochemistry (1 998), 37, 1 77 14-1 7725.). -63- 200817514 2'-0-propylguanosine, adenosine, cytidine, uridine can be found in the literature (Lesnik, Ε·Α· et al. Biochemistry (1 993), 3 2, 7 832-78 3 8 ·)synthesis. 2 allyl guanosine, adenosine, cytidine, uridine can be used as a commercially available reagent. 2^0-methoxyethylguanosine, adenosine, cytidine, uridine can be patented (US626 1 840) or literature (Martin, P. Helv. Chim. Acta. (1 995) 7 8, 486- 504.) Synthesis. 2'-〇-butylguanosine, adenosine, cytidine, uridine can be synthesized according to the literature (LeSnik, E.A. et al. Biochemistry (1 993), 32, 7 8 3 2-7 8 3 8.). Pentoguanosine, adenosine, cytidine, and uridine can be synthesized according to the literature (LeSink, E.A. et al. Biochemistry (1 993), 32, 783 2-783 8.). 2 Propargyl guanosine, adenosine, cytidine, uridine can be used as a commercially available reagent. 2'-〇-allyl guanosine, adenosine, cytidine, uridine can be used as a commercially available reagent. 2'-〇, 4'-C-methylene guanosine, adenosine, 5-methylcytidine and thymidine can be according to the method of WO 99/14226, the carbon number of the alkyl group is 2-5 Y- 〇, 4, CM alkyl guanosine, adenosine, 5-methyl cytidine and thymidine can be produced according to the method of WOOO/475 99. When the phosphoric acid group is thiolated, it can be reacted with trivalent phosphorous acid to form a sulfuric acid ester-based reagent, sulfur, tetraethyl thiuram disulfide (TETD, Applied Biosystems), Beaucage reagent (Glen Research), gasified xanthan gum, etc., obtained by the method of Tetrahedron Letters, 32, 3005 (1991) J. Am. Chem. Soc., 112, 1253 (1990) Ester derivatives. The control pore glass (CPG) used in the synthesizer is a combination of 2·-0-methyl nucleosides and is commercially available. Also methylene guanosine, adenosine, 5-methylcytidine-64 - 200817514 and thymidine can be according to the method of W099/14226, the carbon number of the alkyl group is 2-5 2·-〇, 4\C - Alkyl guanosine, adenosine, 5-methylcytidine and thymidine can be produced according to the method of WOOO/47599, according to the literature (Oligonucleotide Synthesis, Edited by MJ Gait, Oxford University Press, 1 984) Combined with CPG. Using a modified CPG (described in Example 1 2 b of JP-A-7-87982), an oligonucleotide in which a 2-ethyl group is bonded to the 3' end can be synthesized. Also using 3'-amino-modifier C3 CPG, 3'-amino-modifier C7 CPG, glyceryl CPG (Glen Research), 3,-specer C3 SynBase CPG 1000, 3'-specer C9 SynBase CPG 1000 (link tech η o 1 ogies), can be synthesized at the 3' end of the combination of a trans-gate phosphate group or an amine alkyl phosphate group. When the antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof is used as a therapeutic and prophylactic agent for the treatment of a disease, the antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof may be used as it is or The pharmacologically acceptable excipients, diluents, and the like are mixed, for example, in the form of a lozenge, a capsule, a granule, a powder, or a syrup, or an injection, a sacrificial agent, a patch, or an external preparation, etc., rather than oral. . These preparations may be excipients (for example, sugar derivatives such as lactose, white sugar, glucose, mannitol, sorbitol; starch derivatives such as corn starch, potato starch, alpha starch, dextrin; cellulose derivatives such as crystalline cellulose; ; gum arabic; polydextrose; organic excipients such as trimeric glucose; and phthalic acid derivatives such as light phthalic anhydride, synthetic aluminum citrate, calcium citrate, magnesium metasilicate aluminate; a salt; a carbonate such as calcium carbonate; an inorganic excipient such as a sulfate such as calcium sulfate; or a slip agent (for example, a stearic acid metal salt such as stearic acid, calcium stearate or hard-65-200817514 magnesium sulphate; Talc; colloidal vermiculite; propolis such as propolis, cetyl wax; boric acid; adipic acid; sulfate such as sodium sulfate; ethylene glycol; fumaric acid; sodium benzoate; DL leucine; fatty acid sodium salt; Sodium sulfate, dodecyl sulfate such as dodecyl magnesium sulfate; phthalic anhydride such as phthalic anhydride or citric acid hydrate; and the above-mentioned starch derivative), a binding agent (for example, hydroxypropylcellulose, hydroxypropylmethylcellulose) , polyvinylpyrrolidone, polyethylene glycol, and before Excipients of the same compound), clumping agents (for example, low-substituted hydroxypropylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium, internal cross-linked carboxymethylcellulose sodium, cellulose derivatives; carboxy Methyl hydrazine powder, sodium carboxymethyl powder, cross-linked polyvinylpyrrolidone and other chemically modified starches and celluloses, emulsifiers (for example, bentonite, colloidal clay such as magnesium aluminum silicate; magnesium hydroxide) , metal hydroxide such as aluminum hydroxide; anionic surfactant such as sodium dodecyl sulfate or calcium stearate; cationic surfactant such as benzyl chloride ammonium chloride; polyoxyethylene alkyl, polyoxyethylene sorbitan fat a non-ionic surfactant such as an acid ester or a sucrose fatty acid ester, or a stabilizer (parabens such as methylparaben or propylparaben; chlorobutanol, benzyl alcohol, phenylethyl alcohol, etc.) Alcohols; benzalkonium chloride; phenols such as phenol and cresol; thimerosal; dehydroacetic acid; and sorbic acid), bridge flavoring agents (such as commonly used sweeteners, sours, spices, etc.), diluents, etc. The additive is produced by a known method. The therapeutic and prophylactic agent of the present invention preferably contains 0.05-5//mole/mi of the antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof, 0.02-10% w/v of a carbohydrate or a polyvalent alcohol, and 0.01-0.4 %w/v pharmacologically acceptable surfactant. A more suitable range of the content of the antisense oligonucleotide of the present invention or its pharmacologically acceptable salt is 0.1-1 //mol/ml. -66 - 200817514 The above carbohydrates are particularly preferred as monosaccharides and/or 2 sugars. Examples of such carbohydrates and polyvalent alcohols may be glucose, galactose, mannose, lactose, maltose, mannitol and sorbitol. These can be used individually or in combination. Further, examples of the surfactant may be polyoxyethylene sorbitan mono-tri-ester, alkyl phenyl polyoxyethylene, sodium taurocholate, sodium cholate, and polyvalent alcohol ester. Among them are polyoxyethylene sorbitan mono-tri-esters, among which ester-specific oleic acid esters, terephthalates, stearates and palmitic acid esters. These can be used individually or in combination. Further, the therapeutic and prophylactic agent of the present invention preferably contains 0.03-0.09 Μ a pharmacologically acceptable neutral salt such as sodium chloride, potassium chloride and/or calcium chloride. Further, the therapeutic and prophylactic agent of the present invention preferably contains 0.002-0.05 of a pharmaceutically acceptable buffer. Suitable examples of buffers are sodium citrate, sodium glycinate, sodium phosphate, and hydroxymethylaminomethane. These buffers can be used singly or in combination. The therapeutic and prophylactic agent of the present invention can also be supplied in a solution state. However, in order to stabilize the antisense oligonucleotide and prevent the therapeutic effect from being lowered, it is usually preferred to freeze the antisense oligonucleotide. In this case, the solution is used as a solution (for injection). Distilled water, etc., is reconstructed, that is, it can be used in a liquid state that can be administered. Therefore, the therapeutic and prophylactic agent of the present invention also includes a lyophilized state which is used to reconstitute the components in a predetermined concentration range. In order to promote the solubility of the lyophilizate, an amino acid such as a protein or a glycine may be further contained. When the antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof is administered to a human, it can be administered, for example, in an adult dose of about 1 to 100 mg/kg (body weight), preferably 1 to 50 mg/kg (body weight). In one or more times, oral administration or intravenous injection, but the amount of administration and -67-200817514 can be changed according to the type, symptoms, age, and administration method of the disease. The antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof can be administered to a patient suffering from rheumatoid arthritis as follows. The antisense oligonucleotide of the present invention or a pharmacologically acceptable salt thereof is also produced by a method known per se, and this is sterilized by a usual method, for example, to prepare an injection solution of 1200 // g/ml. This solution is administered intravenously to the patient so that the administration amount of the antisense oligonucleotide is, for example, 20 mg per kg body weight, for example, in the form of an infusion. The administration is repeated, for example, four times at intervals of one week, and thereafter, the treatment effect is confirmed by using clinical symptoms and tissue as an indicator, and the treatment is appropriately repeated. If there is a therapeutic effect without obvious side effects, continue treatment. EXAMPLES Hereinafter, the present invention will be specifically described by way of examples, reference examples, test examples and formulation examples. These examples and the like are illustrative of the invention and are not intended to limit the scope of the invention. [Example 1] A nucleic acid automatic synthesizer for synthesizing HO-Te2-C-Te2-T-5Ce2-G-Te2-GCTTA-Ge2-G-Ge2-T-5Ce2-Ae2t-H(AS-1-2) (ABI model 394 DNA/RNA synthesizer manufactured by Bajin Emma Co., Ltd.) was carried out according to a procedure of 40 nmol scale. The concentration of the solvent, the reagent, and the amino phosphate in each synthesis cycle is the same as that of the natural oligodeoxynucleotide. Non-natural type of amino phosphate is described in Example 14 of Patent No. 3,420,984 (5·-0-dimethoxytrityl-2, anthracene, 4'-C-extended ethyl-6-N-benzylhydrazine Adenosine-3·-〇-(2-cyanoethyl N,N-diisopropyl)amino phosphate), Example 27 (5·-〇-dimethoxytrityl-68- 200817514 Ethyl 2-N-isobutyl-guanosine-guanosine- 3·-indole-(2-cyanoethyl N,N-diisopropyl)amino phosphate), Example 22 (5'-〇-dimethyl Oxytrityl l-HC-extended ethyl 4-N-benzylidene-5-methylcytidine-3^0-(2-cyanoethyl N,N-diisopropyl)amino Phosphate), Example 9 (5·-〇-dimethoxytrityl-2·-indole, 4'-(:-extended ethyl-5-methyluridine-3^〇-(2) -Cyanoethyl hydrazide small diisopropyl) Amino phosphate. The solid phase carrier was synthesized with Universal-Q-CPG (manufactured by Glen Research) 0.2//mol to synthesize the title compound. The phosphate coupling time is 15 minutes. ^ The protected oligonucleotide having the desired sequence is treated with concentrated aqueous ammonia to cut the oligomer from the support, and the cyanoethyl group with the protecting group of the phosphate group Removal of nucleobases The base is removed. The solvent is evaporated under reduced pressure, and the residual residue is reversed 1^1^ (1^-1 (^?, manufactured by Shimadzu Corporation), column (^^"1^, 〇111'〇111〇1 Discussion 11 Performance RP-18e (4.6xl00 mm)), with A solution: 5% acetonitrile, 0.1 Μ acetic acid triethylamine aqueous solution (TEAA), pH 7.0, B solution: acetonitrile, Β%: 10%-> 45% (8 Fraction, linear gradient); 60 ° 〇; 21111 / min; 26 〇 11111) refined, collect the peak of the target material with dimethoxytrityl group. Add water and distill off under reduced pressure to remove hydrazine. Add 80% acetic acid Aqueous solution (500 / ζ 1), place 20 minutes to deprotect the dimethoxytrityl group. After distilling off the solvent, the residue is dissolved in 500 // 1 water, with a 0.45 // m filter (MILLIPORE, Ultrafree-MC) was filtered to obtain the desired oligonucleotide. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6 x 100 mm)), solution A: 5% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA); ρΗ7·0, B dissolved: 25% acetonitrile, 0·1Μ acetic acid triethylamine aqueous solution (TEAA), ρΗ7.0, Β%··20%-> 80% ( 8 points, -69- 200817514 linear gradient); 60 ° C 2ml/min; 260nm) analysis 'dissolved at 5.22 points (2.62 A26〇 unit), and the compound was identified by negative ion ESI mass analysis (calculation 値: 5 8 6 0 · 9 6, measured 値: 5 8 6 1 · 01). The base sequence of the present compound is a sequence complementary to nucleotide number 564-58 1 of (GenBank No. NM_0 11 06 1.1). [Example 2] The synthesis of HO-Te2-5Ce2-T-Te2-C-Ge2-TGCTTA-Ge2-G-Ge2-T-5Ce2-Ae2t-H(AS-ll) was synthesized in the same manner as the compound of Example 1. The compound of Example 2 of the sequence of interest. This compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)) 'A solution: 5% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (TEAA), ρΗ7· 0, Β Solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (ΤΕΑΑ), ρΗ7·0, Β%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm) For analysis, it was dissolved at 5.22 minutes (2,63 A26 units), and the compound was identified by negative ion ESI mass analysis (calculation 586: 5609.96, measured 値: 5 86 1.0 1 ). The base sequence of the present compound is a sequence complementary to nucleotide number 564-5 8 1 of (Genebank Genebank No. NM.01 1061.1). [Example 3] Synthesis and Example 1 of HO-Ae2-C-Ce2-TC-Ae2-C-Ae2-CTGTC-Te2-T-Ge2-GA-Ae2-CA e2t-H (AS-2-3) The compound of Example 3 was also synthesized as the compound. After deprotection, reverse phase HPLC (LC-70-200817514 10VP, manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6 x 100 mm)) 'A solution: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7.0, B solution: acetonitrile, 45% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm) refined, collected peaks with dimethoxytrityl . Add water and distill off under reduced pressure to remove TEAA. Add 80% acetic acid in water (500 //1) and place for 20 minutes to remove the dimethoxytrityl group. After distilling off the solvent, it was dissolved in 500 // 1 water, washed with ethyl acetate, and filtered through a 0.45/zm filter (MILLIPORE, Ultrafree-MC) # to obtain the desired oligonucleotide. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7.0. , B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), pH 7. 〇, 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm) analysis, then at 5.41 points Dissolution (1.49 Α 26 units), and the compound was identified by negative ion ESI mass analysis (calculation 672: 6726.5 1, determination 値: 6726.27). The base sequence of the present compound is a sequence complementary to nucleotide number 867 -8 87 of (gene pool number ΝΜ_0 11 06 1.1). [Reference Example 1] Synthesis of HO-Te2-5Ce2-Te2-Te2-CGTGCTTAGG-Ge2-Te2-5Ce2l-Ae2t-H(AS-1) The compound of Reference Example 1 having the desired sequence was synthesized in the same manner as the compound of Example 1. . This compound was subjected to reverse phase HPLC (LC-10VP, manufactured by Shimadzu Corporation).

Column (Merck, Chromolith Performance RP-18e (4.6xl00mm)), A -71- 200817514 Solution: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7.0, B solution: 25% acetonitrile, hydrazine 1Μ aqueous solution of triethylamine acetate (ΤΕΑΑ), ρΗ7.0, Β%: 20%-> 80% (8 points, linear gradient); 60°C; 2ml/min; 260 nm) analysis, dissolution at 5.52 (3.19 A26〇 unit) 'The compound was identified by negative ion ESI mass spectrometry (calculation 値: 5 860.88 'measured 値: 5 860.3). The base sequence of the present compound is a sequence complementary to nucleotide number 564-5 8 1 of (gene pool number ΝΜ_0 11 06 1.1). [Reference Example 2] Synthesis of HO-Ae2-C-Ge2-T-5Ce2-A-5Ce2-ACTGTCT-Te2-G-Ge2A-Ae2-C-Ae2t-H (AS-2-2) and Example 1 The compound was also synthesized as a compound of Reference Example 2 having the desired sequence. After deprotection, reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6 x 100 mm)), solution A: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7 .0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), ρΗ7·0, hydrazine solution: acetonitrile, Β%: 10% - 45% (8 points, linear gradient); 60 ° C; 2 ml/min; 260 nm) refined, and the peak of the target having dimethoxytrityl group was collected. Water was added to remove TEAA under reduced pressure. A solution of 80% acetic acid in water (500 μl) was placed for 20 minutes to effect deprotection of dimethoxytrityl. After distilling off the solvent, the residue was dissolved in 500 // 1 water, and filtered through a 0.45 // m filter (MILLIPORE, Ultrafree MC) to obtain the desired oligonucleotide. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6 x 100 mm)), A solution: 5% -72-200817514 acetonitrile, 0.1% acetic acid triethylamine aqueous solution (TEAA) ), pH 7.0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (ΤΕΑΑ), ρΗ7·0, Β%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm) analysis, dissolved at 5.23 (2.14 A26 〇 unit), and the compound was identified by negative ion ESI mass analysis (calculation 67: 67 54.57, measured 値: 6754.2 1). The base sequence of the present compound is a sequence complementary to nucleotide number 867-8 87 of (gene pool number ΝΜ_0 11 06 1.1). [Reference Example 3] Synthesis and Example 1 of H〇-Te2-C-Te2-TC-Ge2-TG-5Ce2-Te2-TA-Ge2-GG-Te2-C-Ae2t-H(AS-1-3) The compound of Reference Example 3 having the desired sequence was also synthesized. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-1 8e (4.6 x 1 00 mm)), solution A: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA) , pH 7.0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), pH 7.0, B%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; For the 260 nm) analysis, it was dissolved at 5.28 (9.22 A26〇 units) and the compound was identified by negative ion ESI mass analysis (calculation 5: 5 846.94, measured 値: 5 84 6.97). The base sequence of the present compound is a sequence complementary to nucleotide number 564-5 8 1 of (GenBank No. NM.01 1061.1). [Reference Example 4] Synthesis of HO-Te2-Ge2-Ae2-5Ce2-CCTAAGCACG-Ae2-Ae2-G-Ae2lH(S1) Compound of Reference Example 4 having a sequence not labeled with PADI4 mRNA-73-200817514 The compound of Example 1 was also synthesized. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7. 0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), ρ Η 7 · 0, 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm) analysis, then at 7.77 Dissolution (8.33 A26 units), and the compound was identified by negative ion ESI mass analysis (calculation 値: 5 85 1.90, determination 値J 852.9). The base sequence of the present compound is the sequence of nucleotide number 564-5 8 1 (gene bank code ΝΜ_0 11 06 1.1). [Reference Example 5] Synthesis of H〇-Te2-Ge2A-5Ce2-C-5Ce2-TAAGCA-5Ce2-G-Ae2-A-Ge2-Ae2t-H(S11) - Reference to a sequence not labeled with PADI4 mRNA The compound of Example 5 was synthesized in the same manner as the compound of Example 1. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (TEAA), pH 7.0. , Β solution: 25% acetonitrile, 0.1M acetic acid triethylamine aqueous solution (ΤΕΑΑ), ρΗ7·0, B%: 20% - 80% (8 points, linear gradient); 60 ° C; 2ml / min; 260nm) analysis Then, it was dissolved at 4.99 minutes (4.84A26〇 unit), and the compound was identified by negative ion ESI mass analysis (calculation 588: 5880.03, determination 値: 5880.04). The base sequence of the present compound is the sequence of nucleotide number 564-5 8 1 (genome number ΝΜ-0 11 061.1). -74- 200817514 [Reference Example 6] The synthesis of HO-Te2-G-Ae2-C-5Ce2-C-Te2-AAGCA-5Ce2-G-Ae2-A-Ge2-Ae2t-H(Sl-2) The compound of Reference Example 6 in which the PADI4 mRNA is the target sequence was synthesized in the same manner as the compound of Example 1. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 0.1% acetic acid triethylamine aqueous solution (TEAA), ρΗ7 ·0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (ΤΕΑΑ), ρΗ7·0, B%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm The analysis was carried out at 5.20 minutes (6.72 A26〇 units), and the compound was identified by negative ion ESI mass analysis (calculation 5: 5 866.00, measured 値: 5 866.06). The base sequence of the present compound is the sequence of nucleotide number 564-5 8 1 (genome number ΝΜ_0 11 06 1.1). [Reference Example 7] (S-1-3) synthesis of H〇-Te2-G-Ae2-CC-5Ce2-TA-Ae2-Ge2-C-5Ce2-GA-Ae2-G-Ae2l-H has not been PADI4 The compound of Reference Example 7 in which the mRNA was the target sequence was synthesized in the same manner as the compound of Example 1. This compound is a reverse phase Η PLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6xl00mm)), A solution: 5% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (TEAA), ρΗ7 .0, Β solution: 25% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (ΤΕΑΑ), ρΗ7·0, Β%: 20% - 80% (8 points, linear gradient); 60 ° C; 2ml / min; 2 60nm) analysis, dissolution at 5.33 (8·56 Α26. -75- 200817514 • unit), and the compound was identified by negative ion ESI mass analysis (calculation 値: 5 8 6 6.0 0, determination 値: 5 8 6 6.0 4). The base sequence of the present compound is the sequence of nucleotide number 564-5 8 1 (genbank number NM_01 1061.1). [Reference Example 8] Synthesis of HO-Ae2-5Ce2-Ge2-Te2-CACACTGTCTTGG-Ae2-Ae2-5Ce2-Ae2t-H (AS-2) The same as the compound of Example 1, the reference Example 8 f having the desired sequence was synthesized. Compound. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7. 0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), ρΗ7·0, Β%: 20%-> 80% (10 points, linear gradient); 60 ° C; 2 ml / min; The 260 nm) analysis was eluted at 6.96 minutes (8.40 A26 units), and the compound was identified by negative ion ESI mass analysis (calculation 値:6754.57, measured 値: 675 5.9). The base sequence of the present compound is a sequence complementary to the nucleotide number 867 -8 87 of (Genebank No. NM_01106 1.1). [Reference Example 9] The synthesis of H〇-Ae2-5Ce2-GT-5Ce2-Ae2-CACTGTCTT-Ge2-Ge2-AA-5Ce2-Ae2-H (AS-2-l) has the same sequence of interest as the compound of Example 1. The compound of Reference Example 9 was synthesized in the same manner as the compound of Example 1. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6 x 100 mm)), solution A: 5% acetonitrile, 0.1 Μ • 76-200817514 aqueous solution of triethylamine acetate (TEAA) , ρΗ7·0, B solution: 25% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (TEAA), ρΗ7.0, Β%: 20%-> 80% (8 points, linear gradient); 60 ° C; 2 ml/min; 260 nm) was eluted at 5.27 minutes (3.95 A26 units), and the compound was identified by negative ion ESI mass analysis (calculation 676:6768.60, measured 値:6768.20). The base sequence of the present compound is a sequence complementary to nucleotide number 867-887 of (gene bank code ΝΜ_0 11061.1). [Reference Example 10] Synthesis and Examples of HO-Ae2-CG-Te2-CA-5Ce2-AC-Te2-G-Te2-CT-Te2-GG-Ae2-AC-Ae2t-H (AS-2-4) The compound of Example 1 which has the same sequence of interest as the compound of Example 1 was synthesized in the same manner as the compound of Example 1. This compound was subjected to reverse phase HPLC (LC-10VP, manufactured by Shimadzu Corporation, column (Merck, Chromolith)

Performance RP-18e (4.6xl00mm)), A solution: 5% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (TEAA), ρΗ7·0, hydrazine solution: 25% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (ΤΕΑΑ ), ρΗ7·0, Β%: 20%-> 80% (8 points, linear gradient); 60°C; 2ml/min; 260nm) analysis, dissolution at 5.23 minutes (5.31 A26〇 unit), compound It was identified by negative ion ESI mass analysis (calculation 値: 6740.54, measured 値: 6740.39). The base sequence of the present compound is a sequence complementary to nucleotide number 867-887 of (gene bank code ΝΜ_〇11061.1). [Reference Example 11] Synthesis of HO-Te2-Ge2-Te2-Te2-CCAAGACAGTGTG-Ae2-5Ce2-Ge2-Te2t-H(S-2) -77- 200817514 Reference Example 11 having a sequence not labeled with PADI4 mRNA The compound was synthesized in the same manner as the compound of Example 1. However, the use of the literature Bioorg. Med. Chem. (2003) 11, 221 pp 22 26 describes 5·-〇-dimethoxytrityl-2'-indole, 4'-C-extended ethyl-5 - Methyl uridine combined with CPG. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6 x 100 mm)), solution A: 5% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH 7. 0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), pH 7.0, B%: 20%-> 100% (8 points, linear gradient); 60 ° C; 2 ml / min; The 260 nm) analysis was eluted at 6.71 minutes (5.78 A26 〇 unit), and the compound was identified by negative ion ESI mass analysis (calculation 68: 68 1 1.40, measured 値: 6811.9). The base sequence of the present compound is the sequence of nucleotide number 867-887 of (gene bank code NM_01 1061.1). [Reference Example 1 2] The synthesis of H〇-Te2-Ge2-TT-5Ce2-5Ce2_A-AGACAGTG-Te2-Ge2-AC-Ge2-Te2l-H(S2-l) has a sequence not labeled with PAD 14 mRNA The compound of Reference Example 12 was synthesized in the same manner as the compound of Example 1. This compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith)

Performance RP-18e (4.6x100mm)), A solution: 5% acetonitrile, 0.1M aqueous solution of triethylamine acetate (TEAA), pH 7.0, solution B: 25% acetonitrile, 0.1 M aqueous solution of triethylamine acetate (TEAA), pH7.0, B%: 20% - 80% (8 points, linear gradient); 60 ° (:; 21111 / min; 2 6 〇 11111) analysis, then dissolved at 4.98 (1.38 A26. units), and compounds It was identified by negative ion ESI mass spectrometry (calculation -78-200817514 calculus: 6825.60, determination enthalpy: 6825.23). The base sequence of the present compound is the sequence of the nucleotide number 867-8 87 of (gene bank code ΝΜ_01106 1.1). [Reference Example 13] The synthesis of HO-Te2-G-Te2-T-5Ce2-C-Ae2-AGACAGT-Ge2-T-Ge2-A-5Ce2-G-Te2t-H(S-2-2) The compound of Reference Example 12 in which the PADI4 mRNA was the target sequence was synthesized in the same manner as the compound of Example 1. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 0.1% acetic acid triethylamine aqueous solution (TEAA), Η 7.0, Β solution: 25% acetonitrile, 0.1 三 acetic acid triethylamine aqueous solution (TEAA), ρ Η 7.0, Β%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; The 260 nm) analysis was eluted at 5.18 (1.45 A26 〇 unit), and the compound was identified by negative ion ESI mass analysis (calculation 682: 6825.60, measured 値: 6825.20). The base sequence of the present compound is the sequence of nucleotide number 867-8 87 of (Genebank No. NM_01 106 1.1). [Reference Example 14] The synthesis of HO-Te2-G-Te2-TC-5Ce2-A-Ae2-GACAG-Te2-G-Te2-GA-5Ce2-G-Te2t-H(S-2-3) The compound of Reference Example 14 in which the PADI4 mRNA was the target sequence was synthesized in the same manner as the compound of Example 1. This compound was subjected to reverse phase HPLC (LC-10VP, manufactured by Shimadzu Corporation, column (Merck, Chromolith)

Performance RP-18e (4.6x100 mm)), A solution: 5% acetonitrile, 〇·1Μ -79- 200817514 acetic acid triethylamine aqueous solution (TEAA), pH 7.0, B solution: 25% acetonitrile, 0.1 M acetic acid triethyl Amine aqueous solution (TEAA), pH 7.0, B%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm) analysis, dissolution at 5.23 (2.76 Ah. units) Further, the compound was identified by negative ion ESI mass analysis (calculation 682: 6825.60, measurement 値: 6825.49). The base sequence of the present compound is the sequence of nucleotide number 867 -8 87 of (Genebank No. NMJ1 106 1.1). [Reference Example 15] The synthesis of HO-Te2-GT-Te2-CC-Ae2-AG-Ae2-C-Ae2-GT-Ge2-TG-Ae2-CG-Te2t-H (S-2-4) has The compound of Reference Example 15 in which the PADI4 mRNA was the target sequence was synthesized in the same manner as the compound of Example 1. The compound was subjected to reverse phase HPLC (LC-10VP manufactured by Shimadzu Corporation, column (Merck, Chromolith Performance RP-18e (4.6×100 mm)), solution A: 5% acetonitrile, 〇·1Μ acetic acid triethylamine aqueous solution (TEAA), pH 7 .0, B solution: 25% acetonitrile, 0.1 M acetic acid triethylamine aqueous solution (TEAA), pH 7.0, B%: 20% - 80% (8 points, linear gradient); 60 ° C; 2 ml / min; 260 nm The analysis was carried out at 5.40 minutes (2.62 A26〇 unit), and the compound was identified by negative ion ESI mass analysis (calculation 67: 67 9 7.55, determination 値: 6794.46). The base sequence of the present compound is the sequence of nucleotide number 867-887 (gene bank number ΝΜ_0 11061.1). [Reference Example 1 6] Construction of PADI4 gene expression vector of mouse a) First strand cDNA synthesis -80- 200817514 The first strand of cDNA was synthesized by SuperScriptTM II RNase® reverse transcriptase (In vitro gen) as follows. Using 500 ng of pancreatic polya+ RNA (Clontech) as a template, 500 //g/ml oligo p(dT)12-18 cDNA synthesis primer (Roche Diagnostic) 1 was dissolved from 1 in DNase and RNase free water ( Sigma), with a final capacity of 15//1 and a 10 minute hold at 70 °C. It was then transferred to ice to quench, and after standing for 5 minutes, 5 x of the first buffer (added in SuperScriptTM II) 5 μl, 100 mM DTT (added in SuperScripTM II) 2.5/M, 25 mM dNTPs (25 mM each dATP, dCTP, dGTP, dTTP) (Invitrogen) l / / l, 40 units / μ l RN ase inhibitor (Toyobo Co., Ltd.) 0 · 5 // 1, 2 0 0 units / # 1 SuperScriptTM II RNase Η · reverse transcriptase (Invitrogen l//l to modulate 25 // 1 reaction solution. The reaction solution was incubated at 70 ° C for 90 minutes, and after incubated at 70 ° C for 10 minutes, DNase and RNase free water were added to make the total amount 50//1, which was the first cDNA solution. b) Acquisition of the mouse 'd4' cDNA

The first cDNA obtained in a) is the starting material, and the cDNA having the ORF of the PADI4 nucleotide sequence of the mouse according to the following method is used, and the forward primer (5·-TATTAAGCTTCCCTGCTGCCCGAGGATGGCCCAGG-3, (Sequence 歹丨J No. 7), reverse The scorpion scorpion (5,-TATTGCGGCCGCAAGGGAGGCTGCC TGGGGTAGTG-3') (SEQ ID NO: 8) was obtained by using K〇D DNA polymerase (Toyobo Co., Ltd.) according to the protocol to which it was attached. The target cDNA was 1% agarose. The gel was electrophoresed to confirm the increase of the target cDNA (about 2 kbp), and the QIAquick PCR purification kit (QIAGEN) was purified according to the protocol attached thereto. The purified DNA fragment was zero Blunt TOPO PCR -81- 200817514

The pCR-Bluntll-TOPO vector of the Cloning kit (InviUogen) was inserted according to the protocol to which it was added, and the coliform flora containing the plasmid was formed on the acacia medium by the host coliform. The strains were isolated to extract the plasmid, and the plasmid with the DNA insert of about 2 kbp was isolated (Padi4/pCR-Bluntll). The vector for the expression of Padi4 animal cells in mice is continued with forward scorpion scorpion (5: GGGAAGCTTGCCACCATGCATCACCATCACCA τοΑΟοαοοΑΟΟοτοοσστσΑταοΑΟΟτο-β'κ 歹 fj bud squaring number 9), reverse scorpion scorpion (5, CCCGGATCCTCAGTCAGTCAGGGCACCATG 00 0 eight eight eight (:1^八八00丁0八八000(:丁1^(:-3,)(序歹"扁号10), Pfx DNA polymerase (Invitrogen) is attached by Padi4/pCR-BluntII The protocol was used to construct. In the obtained PCR reaction, a DNA fragment of about 2 kbp was specifically amplified by agarose electrophoresis, and the DNA fragment was purified by the QIAquick Gel Extraction Kit (QIAGEN). The DNA fragment was digested with Hind III and Bam HI restriction enzyme in the reaction solution of the added buffer. 'The plasmid vector pCR3.1 (Invhrogen) digested with Hind III and Bam HI was inserted, and the host coliform was formed on the agar medium. Plasmid coliform bacteria. The plasmid is isolated from these floras, and the plasmid having the DNA insert of about 2 kbp is isolated. The whole core of the cDNA inserted into the obtained plasmid. The nucleotide sequence was analyzed by ABI 3700 DNA Sequencer (Applied Bio System) and found to be the following sequence. -82- 200817514

AAGCTTGCCACCATGCATCACCATCACCATCACGCCCAGGGTGCGGTGATC

CACGTGGCCCCCGAGCAGCCCACTCACGCCGTGTGTGTGGTGGGCACCGC

GACCCCGCTGGATGTCCGCGGTTCTGCTCCTAAGGGCTACACAACCTTCAG

CATCACAGCCTCCCCAGGAGTCATCGTAGATGTCACCCATGGTCCTCCAGT

CAAGAAGAGTACCATGGGGGCCTCCAAATGGCCTCTGGACCCTGAGCTGGA

GGTGACCCTACAGGTGAAAGCAGCCAGCAGCAGAACAGATGACGAAAAGGT

TCGAGTTTCATACTATGGACTCAAGACCTCCCCAGTCCAAGCCCTGATCTA

CATCACTGGGGTGGAACTGTCCCTGAGCGCAGATGTCACCCGCACTGGCAG

AGTGAAGCCAGCCCAAGCCGGGAAGGATCAGAGCACCTGGACCTGGGGCC

CGGGCGGCCGTGGCGCCATCCTGTTGGTGAACTGTGACAAAGAGGACCCTC

AGGCCTCCGGAATGGACTTTGAGGATGACAAGATCTTGGACAACAAAGACC

TGCAGGACATGTCTCCAATGACCCTAAGCACGAAGACGCCCAAAGACTTCT

TTGAAAAGTATCAGCTGGTGTTGGAGGTGCCCAAGGCCAAGATGAACAGAG

TGAGAGTCTTCCGGGCCACACGGGGCAAACTGCCGTCCCGGTACAAGGTGG

CCCTGGGACCACAACAGTTCTCGTATTGCCTGGAGCTGCCCGGCGGCCAGC

ACAGCACAGACTTCTATGTGGAAGGCCTTGCTTTCCCAGACGCAGACTTCA

AAGGGCTCATTCCCCTCACCATCTCCCTGCTGGACAAGTCTAACCCGGAGC

TCCCCGAGGCCCTGGTGTTCCAAGACAGTGTGACGTTCCGTGTGGCCCCCT

GGATCATGACCCCCAACACTCAGCCCCCCCAGGAGGTGTACGTGTGCAGGG

TrTCTGACAATGAAGACTTCCTAAAGTCACTAGCTACTCTGACCAAGAAAGC

CAAGTGCAAGCTGACTGTGTGCCCCGAGGAGGAGAATATAGATGACCAATG

GATGCAGGACGAAATGGAGATTGGCTACATCCAGGCCCCACACAAGACGCT

GCCTGTGGTCTTTGACTCCCCGAGGGACAGAGGCCTGAAGGATTTTCCTGT

CAAGCGAGTTATGGGTCCAAATTTTGGCTATGTGACCCGAAAGCTCTATATG

TCAGAGCTCACTGGGCTGGATGCCTTTGGGAACCTGGAGGTGAGTCCCCCA

GTCACTGTCAGAGGGAAGGAGTACCCACTGGGCAGAATTCTCATCGGGAAT

AGCGGTTACTCCAGCAGCGAGAGCCGGGACATGCACCAGGCCCTGCAGGA -83- 200817514

CTTCCTGAGCGCCCAGCAGGTGCAGGCCCCCGTGAGGCTCTTCTCCGATTG

GCTCTTTGTGGGTCACGTGGATGAGTTCTTGAGCTTTGTCCCAGTGAGGGA

CAAGCAGGGTTTTCGGCTGCTGCTGTCCAGCCCCAGAGCTTGCTATCAGCT

GTTCCAGGAGCTACAGAGCCAGGGCCACGGGGAGGCGACACTGTTCGAAG

GACTCAAGAGGAAAAGGCAGACAATCAATGAAATTCTGTCCAACAAGAAAC

TAAGAGACCAGAATGCCTATGTGGAGAGCTGTATCGACTGGAACCGGGCGG

TGCTGAAGCGGGAGCTGGGCCTGGCGGAGGGTGACATCATCGACATCCCG

CAGCTCTTCAAGCTCGTGGGGAACTCCAGAGGGAACTCTAAGGCCCAGGCC

TTCTTCCCAAACATGGTGAACATGCTGGTCCTGGGCAAGTACCTGGGCATC

CCCAAGCCCTTTGGGCCCATCATCGATGGCCGCTGCTGCCTGCAGGAGGAG

GTGCGTTCCCACCTGGAGCCACTGGGTCTGCACTGCACCTTCATCAACGAC

TTCTACACCTACCACGTGTACAACGGGGAGGTTCACTGTGGCACCAATGTG

CGCAGGAAGCCCTTCACCTTCAAGTGGTGGCACATGGTGCCCTGACTGACT GAGGATCC (SEQ ID NO: 11) [Reference Example 17] Preparation of pADI4 cRNA The plasmid (680 // g/mL, 10 β L) having the PADI4 gene of the reference sample prepared in Reference Example 16 was used as a scorpion (GAATTCTAATACGACTCACTATA GGGAGAC ( Preface J No. 12) 10 // M, TGCTGG AT ATCTGC AG AA TTCGGCT (Serial [J No. 13) 10// M) 50// L, with Ex Moq // L (Baosheng Technology Co., Ltd.) ), sterilized water 140 / L, with PCR thermal cycler PERSONAL (Baosheng Technology Co., Ltd.) to increase. After the reaction was incubated at 94 ° C for 10 minutes, the reaction was repeated at 94 ° C for 1 minute, at 63 ° C for 1 minute, and at 72 ° C for 1 minute. After the reaction solution 500 was added with chloroform 25 0 /z L and phenol 25 (L), the mixture was centrifuged to recover an aqueous layer. After ethanol precipitation, it was washed with diethyl ether and dissolved in sterilized water to about 1 ml to be ultrafiltered. The membrane (Microcon, YM-50) was centrifuged, and the filtrate was added with sterilized water 3 50 // L, and then filtered. This operation was repeated 3 times -84-200817514, and the solution in the upper part of the filter was collected in increments of 20 μ of sterilized water. 1. More 4 Μ ammonium acetate aqueous solution 200 / z L, cooled ethanol 1 mL, ethanol precipitation, to prepare a DNA fragment of 0.302 / / g / / / l concentration 100 / / 1 (0.603 OD). From the preparation of DNA fragments with AmpliScdbe The T7 transcript kit modulates cRNA. The DNA fragment solution described above is 6.63 // L 卩 10xT7 reaction buffer 4.0 // L, 100mm ΑΤΡ 3.0 / z L, 100 mM CTP3.0 /z L, lOOmM GTP3.0#L , 100 mM UTP3.0 / / L, 10 mM DTT4.0 / / L, Ampliscribe T7 enzyme solution 4.0 / L, DEPC treated water to make a total amount of 40.0 / z L, after heating for 37t 2 hours, ethanol precipitation. The UV was measured to quantify, and a cRNA of a concentration of 7.33/M was prepared. The cRNA was stored in a freezer and appropriately dissolved for the next reaction. [Test Example 1] Reaction of antisense oligonucleotides with RNase(R) of PADI4 cRNA with about 18 pmol of PADI4 cRNA and 100 pmol of the oligonucleotide synthesized in the examples or reference examples in RNase buffer (40 mM Tns-HCl pH 7.7, 4 mM MgCh) , ImM DTT, 40% glycerol, 0.03% BSA, Baosheng Technology Co., Ltd.) 3.4 / L total, 1U coli from RNase H (1.0 / L, Baosheng Technology Co., Ltd.), 0.1U human placenta origin RNase inhibition 15 minutes at 37 ° C in the presence of a dose (0.1 // L, Bioscientific). Add 5x methotrexate gel buffer (0.1 M MOPS ρΗ7·0, 40 mM sodium acetate, 5 mM ED ΤΑ) 2 / L, 37% formaldehyde 3.5 / L, methotrex 15 / L, heated at 65 ° C for 15 minutes to stop. More loaded solution (50% glycerol, ImM EDTA pH 8.0, 0.25% bromine Blue, 0.25% xylene cyanol FF) 2.0 / / L. Decomposition of cRNA by RNase 溴 bromination with 2% agarose gel containing formaldehyde -85-200817514 (lx MOPS-EDTA, 70V, 2 hours) Ethidium staining, LRNA is resolved by chain length separation. The size of the RNA was used to identify Perfect RNAtm Markers, 0.1-1 kb, manufactured by Novagen Corporation, containing 1000, 800, 600, 400, 300, 200, 100 bases of RNA. The stained gel was visualized using a molecular image FX fluorescence imaging system (manufactured by Bio-Rad), and quantified using Quantity One software (manufactured by Bi〇-Rad). Oligonucleotides for administration in Table 3 (AS-2, AS-2-1, AS-2-2, AS-2-3, AS-24, S-2, S-2-1, S- The sequence of 2-2, S-2-3, S-2-4) (the continuous DNA portion is represented by the 2nd lower line), and the continuous DNA number is integrated. Figure 5 shows the results of gel electrophoresis of the reaction of the oligonucleotide with the RNase 2 of the 2 strands of PADI4 cRNA. The continuous DNA was 13, and AS-2 having an antisense sequence caused cleavage at the main site of interest, and a cut piece of about 1230 bases and 920 bases was observed. Further, a part of the target portion was cut. In the case of a continuous DNA of 13, and having a sense sequence which is not complementary to the PADI4 complementary sequence, a plurality of cut fragments were observed. In the case of AS-2-1, AS-2-2, and AS-2-3 with a continuous DNA number reduced to 9, 7 or 5, the cuts other than the target site were reduced. In the series of S-2 having a sense sequence, in S-2-1, S-2-2, and S-2-3 in which the number of consecutive DNAs is reduced to 9, 7 or 5, the number of consecutive DNAs is not seen. The fragment was cut, especially for continuous S-2-3 with a DNA number of 5, and no cut was observed. In the AS-2-4 and S-2-4 with a continuous DNA number of 2 or less, no cleavage due to RNase was observed. -86- 200817514 Table 3

Name consecutive DNA sequence (5'-3') AS-2 Ί 3 ACG: ^CA CAC TGT CTT GGA ACm AS-2-1 Ό ACG- TGK: CAC TGT CTT GGK ACA AS-2-2 r ' 7 CG CG TCA CAC TGT ςτΓ GGK ACA. AS-2-3 r 5 A C0 TCA CSC TGT C^T gga ACM AS-2-edge ho gap ACG , CA CAC TGT CTT GGA ACA S-2 r ' 13 TGT TCC AAG AC A GTG TGA; COT S - 2M FGT TCC AAG ACA GT〇.TGA COT : S-2-2 Η TG? TCC A A<? ACA gtg TGA CGT:: S-2-3 5 : TGT' TCC AAG AGA QTG •TGA: :CGT - S - 2-4 no gap TCC ACA GTS TGA CGT

MM : ΕΝΑ Non-italic: DNA The oligonucleotides supplied to the experiment in Table 4 (AS-1, AS-ll, AS-1-2, AS-Bu 3, Sl, Sl-1, Sl-2, S The sequence of -2-3) (the continuous DNA portion is represented by the 2-fold lower line), and the continuous DNA number is integrated. Figure 6 shows the results of gel electrophoresis of the reaction of the oligonucleotide with the RNase 2 of the 2 strands of PADI4 cRNA. The continuous DNA was 10, and AS-1 having an antisense sequence caused cleavage at the main target site, and a cut fragment of about 1 540 bases and 610 bases was observed. A part of the cutting except for the target site was observed. A plurality of cut fragments were observed in S1 with a continuous DNA of 10 and having a sense sequence that is not complementary to the PADI4. In the case of AS-1-1, AS-1-2, and the target site where the number of consecutive DNAs was reduced to 6 or 5, the cutting was reduced. In the series of S-2 having a sense sequence, S-1-1 in which the number of consecutive DNAs was reduced to 6 or 5, and sputum in S-1-2 was not observed. The number of consecutive DNAs -87- 200817514 is not observed in AS-2-4 or S-2-4 of 2 or less due to RNase 切断. Table 4 Name Continuous DNA number Sequence σ-3)

AS-1 Ίο TCT TQQ TGC TTA QQG TCA AS-b 1 r 6 TCT TCG TGC TTA GGG TCA AS-1-2 r 5 :TCT TCG TQQ TTA ggg TCA AS-1_3 no _ WCT TCG TGC TTA GQG Τ〇Ά S -1 :'r:: 10 TGA ecc TAA GCA QQA aga: S-1 - 1 kW -6 ::!FGA ccc. TAA GCA AGA: S-1 - 2 r 5 ;;ccc: TAA GCA CGA: .. ASA S-1 a 3 no gap Tm ccc ΤΚΆ GCA CGA AGA

Italic: ΕΝΑ Non-italic: DNA Known from these, when the number of consecutive DNAs is 5 or 6, 'The specific sequence of the target site identified in the oligonucleotide with the antisense sequence' has a sense sequence Non-specific cut-off of total sputum in the oligonucleotide. [Test Example 2] The expression inhibition of PADI4 mRNA by the antisense oligonucleotide was evaluated in NIH3T3 of the fibroblast cell line derived from the fetal mouse, and the transient cells were introduced into the reference sample 16 to have a white mouse PAD. The plasmid of 14 gene, and the antisense oligonucleotide of the example of mouse PADI4Mrna 'to quantify the intracellular mouse PADI4 mRNA' was evaluated by the PA D14 m RNA expression of the antisense oligonucleotide of the example. The amount of inhibition effect is -88-200817514 The plasmid having the PADI4 gene of the mouse, and the introduction of the oligonucleotide of the example or the test example into the cell are carried out by using Superfect Transfection Reagent (QIAGEN) according to the attached protocol. Whole RNA was extracted using the RNeasy 96 kit (QIAGEN) from the cells 24 hours after the introduction, and then the DNase-free kit (AMBION) was used to remove the genomic DNA from the obtained whole RNA. Each is implemented in accordance with the attached protocol. The expression of PADI4 mRNA in mice was quantified using TaqMan (Applied Biotechnology System; Analytical ID: Mm〇〇47 8086_ml), and standardized for PADI4 mRNA expression in mice, using TaqMan ribosomal RNA control reagent (application biotechnology system) To quantify 18S ribosomal ENA (rRNA). The RT-PCR reaction was carried out using a MicroAmp optical 96-well reaction plate (application biotechnology system), and the composition of the reaction solution was as follows. Total RNA solution 2 // 1, 2x — Step RT-PCR Master Mix 25 // 1, 40x Multiscfibe & RNase Inhibitor Mix 1.25// 1. TaqMan Probe 2.5// 1, RNase free water 19.25 μ 1. For the purpose of the calibration curve, only the whole RNA solution from which the PADI4 gene was transiently introduced into the mouse was pre-modulated, and the 5-fold dilution series was repeated, and the dilution series of 625, 125, 25, 1, 0 was inexpensively prepared. Reaction and detection using the ΑΒΙ 7900ΗΤ sequence detection system (application biotechnology system), after reacting at 30 ° C for 30 minutes and 95 ° C for 10 minutes, repeat the reaction at 95 ° C for 10 seconds and 60 ° C for 1 minute. Back, the amount of luminescence of the reporter pigment was measured every one cycle. The amplification curve of PADI4 and rRNA in mice was prepared from the amount of luminescence of the reporter pigment in each cycle. The increase curve of the dilution series of the whole RNA solution is prepared by the calibration line, and the sensitivity is obtained on the horizontal axis. The number of cycles is taken as the calibration curve on the 縦 axis, and each table is 89·200817514. The sample for quantitative use is arbitrarily set in the logarithmic amplification period. The number of cycles exceeding a certain amount of illuminance is determined on the calibration line, and the relative amount of performance is calculated. The performance of PADI4 in mice is corrected by the amount of expression of rRNA in the word-sample. In the introduction of the PADI4 gene and the oligonucleotides (AS-l-3, S11, Sl-2, Sl-3) that are thought to not inhibit the expression of PADI4, the expression of PADI4 mRNA in mice was higher than that of the PADI4 gene alone. Improve performance. The improvement in the amount of expression was compared with the PAD 14 gene alone, and when the oligonucleotide was added, it was considered that the ratio of the cation of the amino group of the r' % dyeing reagent to the anion of the phosphate group of the nucleic acid was changed. Then, the amount of PADI4 mRNA expression when each oligonucleotide was used was calculated by using 100% of the oligonucleotide S-1-3 which does not inhibit the expression of PADI4 mRNA, and the results are shown in Fig. 7. Inhibition of PADI4 mRNA expression was observed in AS-1 with 10 consecutive strands of DNA and an antisense sequence. Inhibition of PADI4 mRNA expression was also observed for S-1 in which the DNA was consecutive 10 and had a sense sequence that was not complementary to PADI4. In contrast, when the number of consecutive DNAs is reduced to 6 or (5, and AS_1-1, AS-1-2 having an antisense sequence, there is inhibition of PADI4 mRNA expression, but the number of consecutive DNA is reduced to 6 or 5, and In S-1-1 and S-1-2 which are not in the sense sequence with the complementary sequence of PAD 14, no inhibition of PADI4 mRNA expression is observed. From these, it is known that when the number of consecutive DN A is 5 or 6, it is reversed. The expression of PADI4 mRNA is inhibited in the oligonucleotide of the sense sequence, and the inhibition of PADI4 mRNA expression is not observed in the oligonucleotide of the control sequence, and the antisense oligonucleotide with high specificity can be designed. 200817514 [Test Example 3] The melting temperature measurement test order has anti-sense oligonucleotides (AS-2, AS-2-1, AS-2-2, AS-2-3, AS-2-4), and The sense oligonucleotide is a complementary sequence (5'-UGU UCC AAG ACA GUG UGA CGU-3·) (Order [J No. 14) RNA oligonucleotide, and the solution for melting temperature (Tm) measurement ( 12.5 mM phosphate buffer (pH 6.8) was dissolved to a final concentration of 0.33 // 调制 to prepare. Add the two-chain solution (3 mL) at 90 ° C for 5 minutes, then cool to room temperature. Circular dichroism dispersion meter (Japan Spectroscopic J-720 type) The sample was placed in a small cell (small cell thickness of 10 mm), and the temperature was raised to 20 ° C to 80 ° C in a Peltier-type thermostat, and the argon roundness at 260 nm was measured at intervals of 0.1 ° C. The oligonucleotide (all DNA: 5, -ACG TCA CAC TGT CTT GGA ACA-3') (SEQ ID NO: 6) having the same sequence as the antisense oligonucleotide is compared with the temperature rise. The midpoint of the transfer of the molar ratio is taken as the melting temperature (Tm). Antisense oligonucleotides (AS-2, AS-2-1, AS-2-2, AS-2-3, AS-2-4) Compared with All DNA, all showed high Tm値. This is shown by introducing 2'-0,4-extended ethyl cross-linked nucleoside into the antisense oligonucleotide to form a stable relationship with the complementary strand RNA. 2 strands. Since the characteristic DNA is not present in AS-2-3 with a continuous DNA number of 5, the sequence specificity of R Nase in AS-2-3 and PADI4 cRNA observed in Test Example 1 is cut off. The reaction is affected by the stability of the 2 strands, which is more important than the number of consecutive DNAs. -91- 200817514 Table 5 Number of consecutive DNAs of antisense oligonucleotides Tm(°C) All DNA 21 49 AS-2 13 64 AS- 2-1 9 69 AS-2-2 7 70 AS-2-3 5 68 AS-2-4 Μ j\\\ 71 (Formulation Example 1) A soft capsule is prepared by adding a mixture of the compound of Example 1 in a digestive oil such as soybean oil, cottonseed oil or olive oil, and is injected into gelatin with a positive pressure pump to obtain 100 mg of the active ingredient. The soft capsules are washed and dried. (Formulation Example 2) Tablets Manufactured by the usual method, 100 mg of the compound of Example 1, 0.2 mg of colloidal cerium oxide, 5 mg of magnesium stearate, 275 mg of microcrystalline cellulose, llmg of starch and 98.8 mg of lactose were used. Apply the coating skin as expected. (Formulation Example 3) The suspension was prepared to contain 100 mg of the compound of Example 1 in 10 mL, 1 mg of sodium carboxymethylcellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol solution (曰本药局) and 〇.〇25mL vanillin. -92-200817514 (Formulation Example 4) Injection 1.5 parts by weight of the compound of Example 1 was stirred in 1% by volume of propylene glycol, and the mixture was made into a predetermined volume with water for injection, followed by sterilization. (Formulation Example 5) Injection of Ν-(α-trimethylammonium ethenyl)-didodecyl-D-glutamic acid quinone chloride (final concentration 30 nmol/mL), dodecylphospholipid choline (final concentration: 60 nmol/mL), dioleyl phospholipid, ethanolamine (final concentration: 60 nmol/mL), and 1.5% by weight of the compound of Example 1 were stirred in 1% by volume of propylene glycol, and then sterilized by using water for injection. To manufacture. All publications, patents, and patent applications cited in this specification are hereby incorporated by reference. Industrial Applicability The present invention provides an antisense oligonucleotide which can specifically cleave the target sequence. The expression of the target RNA can be inhibited by the antisense oligonucleotide of the present invention. The antisense oligonucleotide of the present invention is useful as a medicine for preventing and/or treating a target RNA-related disease. [Simple description of the diagram] Figure 1 shows the action sequence of the antisense oligonucleotide. Figure 2 shows the structure of DNA, RNA, PS ODN and the N and S configuration. Figure 3 is a diagram showing the construction of an RNA-type modified nucleotide used in the antisense method. Figure 4 shows the design of the antisense of the modified nucleotide of the RNA type. Figure 5 shows the results of gel electrophoresis of the reaction of the oligonucleotide with the RNase Η -93- 200817514 of the 2 strands of PADI4 cRNA. Figure 6 shows the results of gel electrophoresis of the reaction of the oligonucleotide with the RNase 2 of the 2 strands of PADI4 cRNA. Figure 7 shows the results of inhibition of PADI4 mRNA expression by the mouse of the oligonucleotide. Sequence Listing <SEQ ID NO: 1 > SEQ ID NO: 1 is the base sequence of mouse PADI4 mRNA (registered in EMBL/Genebank No. NM_011061). <SEQ ID NO: 2 > SEQ ID NO: 2 is an amino acid sequence encoded by the mouse PADI4 gene (encoded by the nucleotide sequence of SEQ ID NO: 1). <SEQ ID NO: 3> SEQ ID NO: 3 is the nucleotide sequence of human PADI4 mRNA (registered in EMBL/Genebank No. NMJ12387). <SEQ ID NO: 4> SEQ ID NO: 4 is an amino acid sequence (encoded by the nucleotide sequence of SEQ ID NO: 3) encoded by the human PADI4 gene. <SEQ ID NO: 5 > SEQ ID NO: 5 is a sequence complementary to nucleotide number 564-58 1 of the base sequence of mouse PADI4 mRNA (registered in EMBL/Genebank No. NM_011061). <SEQ ID NO: 6 > SEQ ID NO: 6 is a sequence complementary to nucleotide number 867-887 of the base sequence of mouse PADI4 mRNA (registered in -94-200817514 EMBL/Genebank No. NM-111061). <SEQ ID NO: 7> SEQ ID NO: 7 is a sequence in which the primer was used before the reference example 16. <SEQ ID NO: 8 > SEQ ID NO: 8 is the sequence of the reverse primer used in Reference Example 16. <SEQ ID NO: 9> SEQ ID NO: 9 is a sequence in which the reference primer was used before. <SEQ ID NO: 1> The sequence number 10 is the sequence of the reverse primer used in Reference Example 16. <SEQ ID NO: 11> - SEQ ID NO: 1 1 is the sequence 歹 ij of the mouse Padi4 cDNA obtained in Reference Example 16. <SEQ ID NO: 1 2 > SEQ ID NO: 1 2 is the sequence of the primer used in Reference Example 17. <SEQ ID NO: 13> Sequence number 13 is the sequence of the primer used in Reference Example 17. <SEQ ID NO: 1 4 > SEQ ID NO: 14 is the sequence of the RN A oligonucleotide used in Test Example 3 -95-

Claims (1)

200817514 X. Patent application scope: 1. An antisense oligonucleotide of the following sequence (I) or a pharmacologically acceptable salt thereof, Wing1-R11-Window-R12-Wing2 (I) (in sequence (I), Window is a nucleus a deoxyribonucleotide sequence of 5 or 6 nucleotides, R11 and R12 are each a ribonucleotide, and Wing1 and Wing2 are each a ribonucleotide, a ribonucleotide sequence, a deoxyribonucleotide, and an oxygen scavenging a ribonucleotide sequence or a ribonucleotide and (&lt; a mixed sequence of deoxyribonucleotides, Wing1 is a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides, The sequence of deoxyribonucleotides is not more than 4 consecutive, Wing2 is a deoxyribonucleotide sequence or a mixed sequence of ribonucleotides and deoxyribonucleotides, in the sequence, deoxyribonucleoside The acid is not more than 4 consecutive, # constituting the sequence of Wing^R&quot; or R12-Wing2, at least one ribonucleotide V is the saccharide 2'-0 and 4'-C is crosslinked by Cm alkyl chain 2. The antisense oligonucleotide of claim 1 or the pharmacologically acceptable salt thereof, the Cm alkyl chain 3. An antisense oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1 or 2, wherein the RNA of the disease-associated gene is the subject of the patent. An antisense oligonucleotide or a pharmacologically acceptable salt thereof, wherein the disease-associated gene is a PADI4 gene. 5. The antisense oligonucleotide of claim 4 or a pharmacologically acceptable salt thereof, 96-200817514, wherein the target RNA is A base sequence 歹 ij having the gene bank number NM_011061.1 or NM_0 1 23 87. 6. A composition comprising the antisense oligonucleotide of claim 1 or 2 or a pharmacologically acceptable salt thereof. The composition of claim 6 is used as a medicine. 8. The composition of claim 6 is used as a reagent. 9. A method for inhibiting the performance of the target RNA is used as An antisense oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1 or 2. 10. A method for preventing and/or treating a diseased RNA and using the method as claimed in claim 1 or 2 Antisense oligonucleotide or pharmacologically acceptable salt thereof 1 1 · The use of the antisense oligonucleotide of claim 1 or 2 or the pharmacologically acceptable salt thereof is used to manufacture a medicament for the prevention and/or treatment of the target and the disease. -97- 200817514 Sequence Listing &lt;110&gt; First Tri-Company (&lt;120> antisense oligonucleotide with sequence specificity <130> FP-093PCT &lt;150&gt; JP Ρ2006-242403 &lt;151> 2006-09-07 &lt;160&lt;17&gt;&lt;17&gt; Patent version 3.1 &lt;210&gt; 1 &lt;211> 2276 &lt;212&gt; RNA &lt;213> mouse&lt;400&gt; 1 tccctgctgc ccgaggatgg ccoagggtgc ggtgatcaac gtggcccccg agcagcccac tcacgccgtg tgtgtggtgg gcacagcgac cccgctggat gtccgcggtt ctgctcctaa gggctacaca accttcagca tcacagcctc tccaggagtc atcgtagatg tGatccatgg tcctccagtc aagaagagta ccatgggggc ctccaaatgg cctctggacc ctgagctgga ggtgacccta caggtgaaag cagccagcag cagaacagat gaccaaaagg ttcgagtttc atactatgga cccaagacct ccccagtcca agccctgatc tacatcactg gggtggaaot gtccctgagc gcagatgtca occgcactgg cagagtgaag ccagccccag ocgggaagga tcagagcacc tggacctggg gcccgggcgg ccgtggcgca atcctgtt gg tgaactgtga caaagaggac cctcaggcct ccggaatgga ctttgaggat gacaagatct tggacaacaa agacctgcag gacatgtctc caatgaccct aagcacgaag acgcccaaag acttctttga aaagtatcag ctggtgttgg aggtgcccaa ggccaagatg aacaaagtga gagtcttccg ggccacacgg ggcaaactgc tgtcccggta caaggtggcc ctgggaccac aacagttctc gtattgcctg gagctgcccg gcggccagca cagcacagac ttctatgtgg aaggtcttgc tttcccagac gcagacttca aagggctcat tcccctcacc atctccctgc tggacaagtc 60 120 180 240 300 360 420 480 540 600 660 720 780 840 taacccggag ctccccgagg ccctggtgtt ccaagacagt gtgacgttcG gtgtggcccc ctggatcatg acccccaaca ctcagccccc ccaggaggtg tacgtgtgca gggtttctga caatgaagac ttcctaaagt cactagctac tctgaccaag aaagccaagt gcaagctgac tgtgtgcccc gaggaggaga atatagatga ccaatggatg caggacgaaa tggagattgg ctacatGcag gccccacaca agacgctgcc tgtggtcttt gactccccga gggacagagg cctgaaggat ttccctgtca agcgagttat gggtccaaat tttggctatg tgacccgaaa gctctatatg tcagagctca ctgggctgga tgcctttggg aacctggagg tgagtccccc agtcactgtc agagggaagg agtacccact gggcagaatt ctcatcggga atagcggtta Ctc cagcagc gagagccggg acatgcacca ggccctgcag gacttcctga gcgcccagca ggtgcaggcc cccgtgaggc tcttctccga ttggctcttt gtgggtcacg tggatgagtt cttgagcttt gtcccagtga gggacaagca gggttttcgg ctgctgotgt ccagccccag agcttgctat oagctgttcc aggagctaca gagccagggc cacggggagg cgacaotgtt cgaaggactc aagaggaaaa ggcagacaat caatgaaatt ctgtccaaca agaaactaag agaocagaat gcctatgtgg agagctgtat cgactggaac cgggcggtgc tgaagcggga gctgggcctg gcggagggtg aaatcatcga catcocgcag ctcttcaagc tcgtggggaa ctccagaggg aactctaagg cccaggcctt cttcccaaac atggtgaaca tgctggtoct gggcaagtac ctgggcatcc ccaagccctt tgggcccatc atcgatggcc gctgctgcct ggaggaggag gtgcgttccc acotggagco actgggtctg oactgcaGct tcatcaacga cttotacacc taccacgtgt acaacgggga ggttcactgt ggcaccaatg tgcgcaggaa gcccttcacc ttcaagtggt ggcacatggt gccctgagtc actaccccag gcagcctccc ttgaggactg tcctggccag aggtggctgc cccgaccccc accccacccc ccgcagcgag gtgcagcaag agatctgggg atggttgggc ttccacagag gtggccaGtg tccctgctgt gacatgtttg cttcacccac atctggctcc tgtcccatct ctgtgtgtcc tcaaatggtc cttgcatagc a Agctcagct ctgcttagag atgctgcaat aaagatgtag ttttgt 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2276 200817514 &lt;210> 2 &lt;211> 666 &lt;212&gt; PRT &lt;213> small White Rat &lt;400&gt; 2 Met Ala Gin Gly Ala Val Me His Val Ala Pro Giu Gin Pro Thr His 1 5 10 15 Ala Val Cys Val Val Qly Thr Ala Thr Pro Leu Asp Val Arg Gly Ser 20 25 30 Ala Pro Lys Gly Tyr Thr Thr Phe Ser Me Thr Ala Ser Pro Gly Val 35 40 45 f lie Val Asp Val Ile His Qly Pro Pro Val Lys Lys Ser Thr Met Gly 50 55 60 Ala Ser Lys Trp Pro Leu Asp Pro 6lu Leu Glu Val Thr Leu Gin Val 65 70 75 80 Lys Ala Ala Ser Ser Arg Thr Asp Asp Gin Lys Val Arg Val Ser Tyr 85 90 95 Tyr Gly Pro Lys Thr Ser Pro Vat Gin Ala Leu lie Tyr He Thr Gly 100 105 110 Val Glu Leu Ser Leu Ser Ala Asp Val Thr Arg Thr Gly Arg Val Lys 115 120 125 Pro Ala Pro Ala Gly Lys Asp Gin Ser Thr Trp Thr Trp Sly Pro Gly 130 135 140 Gly Arg Giy Ala Me Leu Leu Val Asn Cys Asp Lys Glu Asp Pro G In 145 150 155 160 200817514 Ala Ser Gly Met Asp Phe Glu Asp Asp Lys Me Leu Asp Asn Lys Asp 165 170 175 Leu Gin Asp Met Ser Pro Met Thr Leu Ser Thr Lys Thr Pro Lys Asp 180 185 190 Phe Phe Glu Lys Tyr Gin Leu Val Leu Glu Val Pro Lys Ala Lys Met 195 200 205 Asn Lys Val Arg Va! Phe Arg Ala Thr Arg Qly Lys Leu Leu Ser Arg 210 215 220 Tyr Lys Val Ala Leu Qly Pro Gin Gin Phe Ser Tyr Cys Leu Qlu Leu 225 230 235 240 Pro Gly Gly Gin His Ser Thr Asp Phe Tyr Val Glu Gly Leu Ala Phe 245 250 255 Pro Asp Ala Asp Phe Lys Sly Leu He Pro Leu Thr He Ser Leu Leu 260 265 270 Asp Lys Ser Asn Pro Glu Leu Pro Glu Ala Leu Val Phe Gin Asp Ser 275 280 285 Val Thr Phe Arg Val Ala Pro Ding lie Met Thr Pro Asn Thr Gin Pro 290 295 300 Pro Gin Glu Val Tyr Val Cys Arg Val Ser Asp Asn Glu Asp Phe Leu 305 310 315 320 Lys Ser Leu Ala Thr Leu Thr Lys Lys Ala Lys Cys Lys Leu Thr Val 325 330 335 Cys Pro 6iu Glu Glu Asn He Asp Asp Gin Trp Met Gin Asp 6lu Met 340 345 350 200817514 Glu Ile Gly Tyr Ile Gin Ala Pro His Lys Thr Leu Pro Val Val Phe 355 360 365 Asp Ser Pro Arg Asp Arg Gly Leu Lys Asp Phe Pro Va! Lys Arg Val 370 375 380 Met Qly Pro Asn Phe Gly Tyr Val Thr Arg Lys Leu Tyr Met Ser Qlu 385 390 395 400 Leu Thr Gly Leu Asp Ala Phe Gly Asn Leu Glu Val Ser Pro Pro Val 405 410 415 Thr Val Arg Gly Lys Glu Tyr Pro Leu Gly Arg lie Leu lie Gly Asn 420 425 430 Ser Gly Tyr Ser Ser Ser Glu Ser Arg Asp Met His Gin Ala Leu Gin 435 440 445 Asp Phe Leu Ser Ala Gin 6ln Val Gin Ala Pro Val Arg Leu Phe Ser 450 455 460 Asp Trp Leu Phe Val Gly His Val Asp Glu Phe Leu Ser Phe Val Pro 465 470 475 480 Val Arg Asp Lys 6ln Giy Phe Arg Leu Leu Leu Ser Ser Pro Arg Ala 4B5 490 495 Cys Tyr Gin Leu Phe Gin Glu Leu Gin Ser 6ln Gly His Gly Glu Ala 500 505 510 Thr Leu Phe Glu Gly Leu Lys Arg Lys Arg Gin Thr lie Asn Glu He 515 520 525 525 Leu Ser Asn Lys Lys Leu Arg Asp Qln Asn Ala Tyr Val Glu Ser Cys 530 535 540 200817514 He Asp Trp Asn Arg Ala Val Leu Lys Arg Slu Leu Gly Leu Ala Slu 545 550 555 560 Gly A Sp Me Me Asp He Pro Gin Leu Phe Lys Leu Val Gly Asn Ser 565 570 575 Arg Gly Asn Ser Lys Ala Gin Ala Phe Phe Pro Asn Met Val Asn Met 580 585 590 Leu Val Leu Gly Lys Tyr Leu Gly lie Pro Lys Pro Phe Gly Pro lie 595 600 60S lie Asp Gly Arg Cys Cys Leu Glu Glu Glu Vai Arg Ser His Leu Glu f 610 615 620 \ , Pro Leu Gly Leu His Cys Thr Phe Me Asn Asp Phe Tyr Thr Tyr His 625 630 635 640 Val Tyr Asn Qly Glu Val His Cys Gly Thr Asn Val Arg Arg Lys Pro 645 650 655 Phe Thr Phe Lys Trp Trp His Met Val Pro 660 665 &lt;210&gt; 3 &lt;211&gt; 2263 &lt;212>RNA. &Lt; 4O0 &gt; 3 agccagaggg acgagctagc ccgacgatgg cocaggggac attgatccgt gtgaccccag agcagcccac ccatgccgtg tgtgtgctgg gcaccttgac tcagcttgac atctgcagot ctgcccctga ggactgcacg tccttcagca tcaacgcctc cccaggggtg gtcgtggata ttgcccacag ccctccagcc aagaagaaat ccacaggttc ctccacatgg cccctggaGO 300 ctggggtaga ggtgaccctg acgatgaaag cggccagtgg tagcacaggG gaccagaagg 200817514 ttcagatttc atactacgga cccaagactc caccagtcaa agctctactc tacctcaccg 360 cggtggaaat ctcoctgtgc gcagacatca cccgcaccgg caaagtgaag ccaaccagag 420 ctgtgaaaga tcagaggacc tggacctggg gcccttgtgg acagggtgcc atcctgctgg 480 tgaactgtga cagagacaat ctcgaatctt ctgccatgga ctgcgaggat gatgaagtgc 540 ttgacagcga agacctgcag gacatgtcgc tgatgaccct gagcacgaag acccccaagg 600 acttcttcac aaaccataca ctggtgctcc acgtggccag gtctgagatg gacaaagtga 660 gggtgtttca ggccacacgg ggcaaactgt octccaagtg cagcgtagtc ttgggtccca 720 agtggccctc tcactacctg atggtccccg gtggaaagoa caacatggac ttctacgtgg 780 aggocctcgc tttcccggac accgacttcc cggggctGat taccctcacc atctccctgc 840 tggacacgtc caacGtggag ctccccgagg ctgtggtgtt ccaagacagc gtggtcttcc 900 gcgtggcgcc ctggatcatg acccccaaca cccagccccc gcaggaggtg tacgcgtgca 960 gtatttttga aaatgaggac ttcctgaagt cagtgactac tctggccatg aaagccaagt 1020 gcaagctgac catctgccct gaggaggaga acatggatga ccagtggatg caggatgaaa 1080 tggagatcgg ctacatccaa gccccacaca aaacgctgca cgtggtcttc gactctccaa 1140 ggaacagagg cctgaaggag tttcccatca aacgagtgat gggtccagat tttggctatg 1200 taactcgagg gccccaaaca gggggtatca gtggactgga ctcctttggg aacctggaag 1260 tgagcccccc agtc acagtc aggggcaagg aatacccgct gggcaggatt ctcttcgggg 1320 acagctgtta tcccagcaat gacagccggc agatgcacca ggccctgcag gacttcctca 1380 gtgcccagca ggtgcaggcc cctgtgaagc tctattctga ctggctgtcc gtgggccacg 1440 tggacgagtt cctgagcttt gtgccagcac ccgacaggaa gggcttccgg ctgctcctgg 1500 ccagccccag gtcctgctac aaactgttcc aggagcagca gaatgagggc oacggggagg 1560 ccctgctgtt cgaagggatc aagaaaaaaa aacagcagaa aataaagaac attctgtcaa 1620 acaagacatt gagagaacat aattcatttg tggagagatg catcgactgg aaccgcgagc 1680 tgctgaagcg ggagctgggG ctggccgaga gtgacatcat tgacatcccg cagctcttca 1740 200817514 1800 1860 1920 1980 2040 2100 2160 2220 2263 agctcaaaga gttctctaag gcggaagctt ttttccccaa catggtgaac atgctggtgc tagggaagca cctgggcatc cccaagccct catcaacggc cgctgctgcc tggaggagaa ggtgtgttcc ctgctggagc cactgggcct ccagtgcacc ttcatcaacg acttcttcac ctaccacatc aggcatgggg aggtgcactg oggcaccaac gtgcgcagaa agcccttctc cttGaagtgg tggaacatgg tgccctgagc ccatcttccc tggcgtcctc tccctcctgg ccagatgtcg ctgggtcctc tgcagtgtgg caagcaagag tcgggcccgt Ctt tgtgaa tattgtggct ccctgggggc ggccagccct cccagcagtg gcttgcttto ttctcctgtg atgtcccagt ttcccactct gaagatccca acatggtcct agcactgcac actcagttct gctctaagaa gctgcaataa agttttttta agtcactttg tac (·, &lt; 210 &gt; 4 &lt; 211 &gt; 663 &lt; 212 &gt; PRT &lt; 213> human &lt; 400 &gt; 4 Met Ala Gin Gly Thr Leu Ile Arg Val Thr Pro 6lu Gin Pro Thr His 1 5 10 15 Ala Val Cys Val Leu Qly Thr Leu Thr Qln Leu Asp He Cys Ser Ser 20 25 30 Ala Pro 6lu Asp Cys Thr Ser Phe Ser lie Asn Ala Ser Pro Gly Val 35 40 45 Val Val Asp lie A!a His Ser Pro Pro Ala Lys Lys Lys Ser Thr Gly 50 55 60 Ser Ser Thr Trp Pro Leu Asp Pro Sty Val Glu Vai Thr Leu Thr Met 65 70 75 80 Lys Ala Ala Ser Gly Ser Thr Gly Asp Gin Lys Val Gin lie Ser Tyr 200817514 85 90 95 Tyr Sly Pro Lys Thr Pro Pro Val Lys Ala Leu Leu Tyr Leu Thr Ala 100 105 110 Val Glu Ile Ser Leu Cys Ala Asp Ile Thr Arg Thr Gly Lys Va! Lys 115 120 125 Pro Thr Arg Ala Val Lys Asp 6ln Arg Thr Trp Thr Trp Gly Pro Cys 130 135 140 Gly G!n Gly Ala lie Leu Leu Val Asn Cy s Asp Arg Asp Asn Leu Glu 145 150 155 160 Ser Ser Ala Met Asp Cys Glu Asp Asp Glu Val Leu Asp Ser Glu Asp 165 . 170 175 Leu Gin Asp Met Ser Leu Met Thr Leu Ser Thr Lys Thr Pro Lys Asp 180 185 190 Phe Phe Thr Asn His Thr Leu Val Leu His Val Ala Arg Ser Glu Met 195 200 205 Asp Lys Val Arg Val Phe Gin Ala Thr Arg Sly Lys Leu Ser Ser Lys 210 215 220 Cys Ser Val· Val Leu Gly Pro Lys Trp Pro Ser His Tyr Leu Met Va! 225 230 235 240 Pro Gly Gly Lys His Asn Met Asp Phe Tyr Val Giu Ala Leu Ala Phe 245 250 255 Pro Asp Thr Asp Phe Pro Gly Leu Me Thr Leu Thr Me Ser Leu Leu 260 265 270 Asp Thr Ser Asn Leu Giu Leu Pro Glu Ala Vai Val Phe Gin Asp Ser 200817514 275 280 285 Vai Val Phe Arg Val Ala Pro Trp lie Met Thr Pro Asn Thr Gin Pro 290 295 300 Pro 6ln Slu Val Tyr Ala Cys Ser He Phe Glu Asn Glu Asp Phe Leu 305 310 315 320 Lys Ser Val Thr Thr Leu Ala Met Lys Ala Lys Cys Lys Leu Thr He 325 330 335 Cys Pro Glu Glu Glu Asn Met Asp Asp Gin Trp Met Gin Asp Glu Met 340 345 350 Glu Ile Gly T Yr Ile 6!n Ala Pro His Lys Thr Leu Pro Val Val Phe 355 360 365 Asp Ser Pro Arg Asn Arg Gly Leu Lys Glu Phe Pro lie Lys Arg Val 370 375 380 Met Gly Pro Asp Phe 6ly Tyr Val Thr Arg Gly Pro Gin Thr Gly Gly 385 390 395 400 lie Ser Gly Leu Asp Ser Phe Gly Asn Leu Gtu Val Ser Pro Pro Val 405 410 415 Thr Val Arg Gly Ly$ Glu Tyr Pro Leu Gly Arg Leu Phe G!y Asp 420 425 430 Ser Cys Tyr Pro Ser Asn Asp Ser Arg Gin Met His Gin Ala Leu Gin 435 440 445 Asp Phe Leu Ser Ala Gin Gin Vin Gin Ala Pro Val Lys Leu Tyr Ser 450 455 460 Asp Trp Leu Ser Val Gly His Val Asp Glu Phe Leu Ser Phe Val Pro -10- 200817514 465 470 475 480 Ala Pro Asp Arg Lys 6ly Phe Arg Leu Leu Leu Ala Ser Pro Arg Ser 485 490 495 Gys Tyr Lys Leu Phe Gin 6lu Gin Gin Asn Glu Gly His Gly Glu Ala 500 505 510 Leu Leu Phe Glu Gly Me Lys Lys Lys Lys Gin 6ln Lys lie Lys Asn 515 520 525 Me Leu Ser Asn Lys Thr Leu Arg Glu His Asn Ser Phe Val Glu Arg 530 535 540 Cys lie Asp Trp Asn Arg Glu Leu Leu Lys Arg Glu Leu Gly Leu Ala 5 45 550 555 560 Glu Ser Asp lie He Asp He Pro Gin Leu Phe Lys Leu Lys Glu Phe 565 570 575 Ser Lys Ala Glu Ala Phe Phe Pro Asn Met Val Asn Met Leu Val Leu 580 585 590 Gly Lys His Leu Gly lie Pro Lys Pro Phe Gly Pro Va! lie Asn Gly 595 600 605 Arg Cys Cys Leu Glu Glu Lys Val Cys Ser Leu Leu Qlu Pro Leu Giy 610 615 620 Leu Sin Cys Thr Phe Ile Asn Asp Phe Phe Thr Tyr His Ile Arg His 625 630 635 640 Gly Glu Val His Cys Gly Thr Asn Val Arg Arg Lys Pro Phe Ser Phe 645 650 655 Lys Trp Trp Asn Met Val Pro -11- 660 200817514 &lt;210&gt; 5 &lt;211&gt; 18 &lt;212&gt; DNA/RNA &lt;213>Manual&lt;220&gt; Antisense oligonucleotide marker mouse PAD 14 mRNA &lt;400> 5 · tcttcgtgct tagggtca &lt;210> 6 &lt;211> 21 &lt;212&gt; DNA/RNA &lt;213>manual&lt;220&gt; 〉Antisense oligodeoxynucleotide flag PAD 14 mRNA &lt;400> 6 21 acgtcacact gtcttggaac a siAX &lt;210&gt;&lt;211&gt;&lt;212〉&lt;213〉&lt;220>&lt;223>Introduction&lt;223&gt;;_&gt; 7 , tattaagctt ccctgctgcc cgaggatggc Gcagg &lt;210〉 8 , &lt;211> 35 &lt;212> DNA &lt;213&gt; Labor &lt;220&gt;&lt;223> Introduction &lt;400> 8 -12· 200817514 tattgcggcc gcaagggagg Ctgcctgggg tagtg 35 &lt;210> 9 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213 Labor &lt;220> &lt;223>Introduction&lt;400> 9 gggaagcttg ccaccatgca tcaccatcac catcacgccc agggtgcggt gatccacgtg 60 &lt;210&gt; 10 &lt;211&gt; 58 &lt;212> DNA &lt;213>Manual&lt;220&gt;&lt;223&gt; primer &lt; 400 &gt; 10 cccggatcct cagtcagtca gggcaccatg tgccaccact tgaaggtgaa gggcttcc 58 &lt; 210> 11 &lt; 2t1 &gt; 2045 &lt; 2t2 &gt; DNA &lt; 213> mouse &lt; 400 &gt; 11 aagcttgcca ccatgcatca ccatcaccat cacgcccagg gtgoggtgat coacgtggcc 60 cccgagcagc ccactcacgc cgtgtgtgtg gtgggcaccg cgaccccgct ggatgtccgc 120 ggttctgctc ctaagggcta cacaaocttc agcatcacag cctccccagg agtcatcgta 180 gatgtcacco atggtcctcc agtcaagaag agtaocatgg gggcctccaa atggcctctg 240 gaccctgagc tggaggtgao cctacaggtg aaagcagcca gcagcagaac agatgacgaa 300 aaggttcgag tttcatacta tggactcaag acctccccag tccaagccct gatctacatc 360 actggggtgg aactgtccct gagcgcagat gtcacccgca ctggcagagt gaagccagcc 420 caagccggga aggatcagag caoctggacc tggggcccgg gcggcogtgg cgccatcatg 480 -13- 200817514 ttggtgaact Gtgaoaaaga ggaccctcag gcctccggaa tggactttga ggatgacaag 540 atcttggaca acaaagacct gcaggacatg tctccaatga ccctaagcac gaagacgccc 600 aaagacttct ttgaaaagta tcagctggtg ttggaggtgc ccaaggccaa gatgaacaga 660 gtgagagtct tccgggocac acggggcaaa ctgccgtccc ggtacaaggt ggccctggga 720 ocacaacagt tctcgtattg Gctggagctg cccggcggco agcacagcac agacttctat 780 gtggaaggcc ttgctttccc agacgcagac ttcaaagggc tcattccoct caccatctcc 840 otgotggaca agtotaaocc ggagotoccc gaggccctgg tgttccaaga cagtgtgacg 900 ttccgtgtgg ccccctggat catgaccccc aacactcagc ccccccagga ggtgtacgtg 960 tgoagggttt ctgacaatga agacttccta aagtcactag ctactctgac caagaaagcc 1020 C aagtgcaago tgactgtgtg ccccgaggag gagaatatag atgaccaatg gatgcaggac 1080 gaaatggaga ttggctaoat ccaggcccca cacaagacgc tgcctgtggt ctttgactcc 1140 ccgagggaca gaggcctgaa ggattttcct gtcaagcgag ttatgggtcc aaattttggc 1200 tatgtgaccc gaaagctcta tatgtcagag ctcaotgggc tggatgcott tgggaacctg 1260 gaggtgagtc ccccagtcac tgtcagaggg aaggagtacc cactgggcag aattctcatc 1320 gggaatagcg gttactccag cagcgagagc cgggac atgc accaggccct gcaggacttc 1380 ctgagcgccc agcaggtgca ggcccccgtg aggotcttct ccgattggct ctttgtgggt 1440 cacgtggatg agttcttgag ctttgtccca gtgagggaca agcagggttt tcggctgctg 1500 ctgtccagcc ccagagcttg ctatcagctg ttccaggagc tacagagcca gggccacggg 1560 u gaggcgacac tgttcgaagg actcaagagg aaaaggcaga caatcaatga aattctgtcc 1620 aacaagaaac taagagacca gaatgcctat gtggagagct gtatcgactg gaaccgggcg 1680 gtgctgaagc gggagGtggg cctggcggag ggtgacatca tcgacatocc gcagctcttc 1740 aagctcgtgg ggaactccag agggaactct aaggcccagg ccttottcoc aaacatggtg 1800 aacatgotgg tcctgggcaa gtacctgggo atccccaagc cctttgggcc catcatcgat 1860 ggccgctgct gcctggagga ggaggtgcgt tcccacctgg agccactggg tctgcactgc 1920 -14- 200817514 1980 2040 2045 accttcatca acgacttcta cacctaccac gtgtacaacg gggaggttca ctgtggcacc aatgtgcgca ggaagccctt caccttcaag tggtggcaca tggtgccctg actgactgag gatcc &lt; 210 &gt; 12 &lt; 211> 30 &lt;212&gt; DNA &lt;213&gt;Labor&lt;220&gt;&lt;223&gt;Introduction&lt;400&gt; 12 30 gaattctaat acgactcact atagggagac &lt;210&gt; 13 &lt;211> 25 &lt;212&gt; DNA &lt;213&gt; Labor &lt;220&gt;&lt;223&gt; Introduction &lt;400> 13 tgctggatat ctgcagaatt cggct 25 &lt;210> 14 &lt;211&gt; 21 &lt;212&gt; RNA &lt;213&gt; Labor &lt;220&gt;&lt;223&gt; RNA oligonucleotide &lt;400&gt; Η uguuccaaga cagugugaog u 21 -15-