リピート結合分子をプローブとしたトリヌクレオチドリピート病の化学生物学研究

2019 Fiscal Year Annual Research Report

• Project/Area Number: 26000007
• Principal Investigator: 中谷 和彦 大阪大学, 産業科学研究所, 教授
• Project Period (FY): 2014 – 2019
• Keywords: トリヌクレオチドリピート / 低分子 / 神経変性疾患 / 遺伝子疾患 / 分子設計 / スクリーニング / 分子合成

Outline of Annual Research Achievements

本研究では、我々の開発するトリヌクレオチドリピート結合分子を用いて、リピート伸長とリピートRNAの機能を低分子で調節する化学を拓き、トリヌクレオチドリピート病の新しい治療法開発に資する創薬リード化合物の創製を目指して、以下4つの研究項目、1)リピート結合分子の性能向上と創製、2)結合分子のリピート不安定化誘導と分子機構の解明・短縮分子探索、3)結合分子によるToxic RNAの捕捉、4)RAN Translationの分子機構解明と低分子による調節原理導出、を実施する。
令和元年度は、研究項目1)に関して、前年度までの成果をもとにCUGリピート結合分子の構造最適化を進め、筋強直性ジストロフィー1型マウスモデルにおいて、顕著なスプライシング改善を示す化合物JM642の創成を達成した。研究項目2および3)では、CAGヘアピンDNAに結合することが判明している化合物NAに、CAG slipped-strand structureへの結合特異性を検証するため、共同研究者であるChristopher Pearson博士とともに、CAG/CTG slipped-strand structure誘導プラスミドをもちいて、EMSAによる検討をおこなった。この結果、CAGリピートが長いほどNAがslip-outへ結合すること、CTG slip-outへはほとんど結合しないことを証明した。さらに、NAの生体でのCAGリピート短縮作用を検討するため、ハンチントン病マウスモデルR6/2の脳線条体組織へ定位脳手術によりNAの直接投与を行った。NAを投与した左側線条体と、生理食塩水を投与した右側線条体で伸長CAGリピート長を比較すると、NA投与側でCAGリピートの短縮がみられた。NAによるハンチントン病モデルマウスでのCAGリピート短縮は、病態の根底にある脳線条体での異常蛋白凝集体を減少させることも実証した。項目4については、新たな化合物CMBL4が、RAN Translationに起因するGFPの生成を抑制する示唆を得た。
以上の研究をもって、特別推進研究の課題に挙げた各項目に関する研究を無事完了した。

Research Products

• [Int'l Joint Research] Scick Kids Institute, Tronto(カナダ)
  • Country Name: CANADA
  • Counterpart Institution: Scick Kids Institute, Tronto
• [Int'l Joint Research] Polish Academy of Sciences, Poznan/Instiute of Bioorganic Chemistry/Adam Mickiewicz University(ポーランド)
  • Country Name: POLAND
  • Counterpart Institution: Polish Academy of Sciences, Poznan/Instiute of Bioorganic Chemistry/Adam Mickiewicz University
• [Journal Article] The dimeric form of 1, 3 - diaminoisoquinoline derivative rescued the mis - splicing of Atp2a1 and Clcn1 genes in myotonic dystrophy type 1 mouse model (2020)
  • Author(s): Matsumoto, J. ; Nakamori, M. ; Okamoto, T. ; Murata, A. ; Dohno, C. ; Nakatani, K.
  • Journal Title: Chem. Eur. J.
  • DOI: 10.1002/chem.202001572
  • Peer Reviewed / Open Access
• [Journal Article] Assembly of ruthenium complexes on double strand DNA using mismatch binding ligand (2020)
  • Author(s): Ni, L. ; Yamada, T. ; Nakatani, K.
  • Journal Title: Chem. Commun. Volume: 56
  • DOI: 10.1016/j.bbrc.2020.03.107
  • Peer Reviewed
• [Journal Article] Recognition of expanded GGGGCC hexanucleotide repeat by synthetic ligand through interhelical binding (2020)
  • Author(s): Lu, Y. ; Dohno, C. ; Nakatani, K.
  • Journal Title: Biochem. Biophys. Res. Commun.
  • DOI: 10.1016/j.bbrc.2020.03.107
  • Peer Reviewed
• [Journal Article] Slipped-CAG DNA binding small molecule induces trinucleotide repeat contractions in vivo (2020)
  • Author(s): Nakamori, M. ; Panigrahi, G. B. ; Lanni, S. ; Gall-Duncan, T. ; Hayakawa, H. ; Tanaka, H. ; Luo, J. ; Otabe, T. ; Li, J. ; Sakata, A. ; Caron, M. C. ; Joshi, N. ; Prasolava, T. ; Chiang, K. ; Masson, J. Y. ; Wold, M. S. ; Wang, X. ; Lee, M. ; Huddleston, J. ; Munson, K. M. ; Davidson, S. ; Layeghifard, M. ; Edward, L. M. ; Gallon, R. ; Santibanez-Koref, M. ; Murata, A. ; Takahashi, M. P. ; Eichler, E. E. ; Shlien, A. ; Nakatani, K. ; Mochizuki, H. ; Pearson, C. E.
  • Journal Title: Nat. Genet. Volume: 52 Pages: 146-159
  • DOI: 10.1038/s41588-019-0575-8
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Novel Naphthyridine Tetramer that Recognizes Tandem G-G Mismatches by Formation of Interhelical Complex (2020)
  • Author(s): Lu, Y. ; Dohno, C. ; Nakatani, K.
  • Journal Title: Chem. Commun. Volume: 56 Pages: 754-757
  • DOI: 10.1039/C9CC08111A
  • Peer Reviewed
• [Journal Article] Structural insights into synthetic ligands targeting A-A pairs in disease-related CAG RNA repeats (2019)
  • Author(s): Mukherjee, S. ; Blaszczyk, L. ; Rypniewski, W. ; Falschlunger, C. ; Micura, R. ; Murata, A. ; Dohno, C. ; Nakatani, K. ; Kiliszek, A.
  • Journal Title: Nucl. Acids Res. Volume: 47 Pages: 10906-10913
  • DOI: 10.1093/nar/gkz832
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] RT-Hpro-PCR : A MicroRNA Detection System Utilizing a Primerwith a DNA Tag (2019)
  • Author(s): Takei, F. ; Akiyama, M. ; Murata, A. ; Sugai, A. ; Nakatani, K. ; Yamashita, I.
  • Journal Title: ChemBioChem Volume: 20
  • DOI: 10.1002/cbic.201900382
  • Peer Reviewed
• [Journal Article] Molecular glue for RNA : Regulating RNA structure and function through a synthetic RNA binding molecule (2019)
  • Author(s): Dohno, C. ; Nakatani, K.
  • Journal Title: ChemBioChem Volume: 20 Pages: 2903-2910
  • DOI: 10.1002/cbic.201900223
  • Peer Reviewed
• [Journal Article] Modulating RNA secondary and tertiary structures by mismatch binding ligands (2019)
  • Author(s): Murata, A. ; Nakamori, M. ; Nakatani, K.
  • Journal Title: Methods Volume: 167 Pages: 78-91
  • DOI: 10.1016/j.ymeth.2019.05.006
  • Peer Reviewed
• [Journal Article] Inhibition of pre-miRNA-136 processing by Dicer with small molecule BzDANP suggested the formation of ternary complex of pre-miR-136-BzDANP-Dicer. (2019)
  • Author(s): Otabe T. ; Nagano K. ; Kawai G. ; Murata A. ; Nakatani K.
  • Journal Title: Bioorg. Med. Chem. Volume: 27 Pages: 2140-2148
  • DOI: 10.1016/j.bmc.2019.03.031
  • Peer Reviewed
• [Presentation] 光異性化特性を向上させた新規光応答性核酸結合分子の開発 (2020)
  • Author(s): 小林笙太朗、堂野主税、中谷和彦
  • Organizer: 日本化学会第100春季年会
• [Presentation] Assembly of ruthenium complexes on dsDNA using mismatch binding ligand (2020)
  • Author(s): Lu Ni, Takeshi Yamada, Kazuhiko Nakatani
  • Organizer: 日本化学会第100春季年会
• [Presentation] Synthesis and properties of 13C-labeled naphthyridine derivative (2020)
  • Author(s): Shuhei Sakurabayashi, Takeshi Yamada, Gota Kawai, Kazuhiko Nakatani
  • Organizer: 日本化学会第100春季年会
• [Presentation] Dicer切断反応を指標としたRNA結合分子標的配列の網羅的解析 (2020)
  • Author(s): 高島 裕介、村田 亜沙子、中谷 和彦
  • Organizer: 日本化学会第100春季年会
• [Presentation] Synthesis and the properties of NA Dimer (2020)
  • Author(s): Chen Qingwen, Yamada Takeshi, Murata Asako, Nakatani Kazuhiko
  • Organizer: 日本化学会第100春季年会
• [Presentation] Synthesis and nucleic acid binding property of a series of quinoxaline-2, 3-dione derivatives (2020)
  • Author(s): Zilu Xing, Tomonori Shibata, Kazuhiko Nakatani
  • Organizer: 日本化学会第100春季年会
• [Presentation] 2-アミノ-1, 8-ナフチリジン二量体の結合位置異性体の合成と核酸との結合評価 (2020)
  • Author(s): 桑原聡己、Das Bimolendu、中谷和彦
  • Organizer: 日本化学会第100春季年会
• [Presentation] Recognition of Pyrimidine Based Asymmetric Internal Loops in Duplex DNA and RNA by ANP77, a Three-Carbon Atom Linked 2-Amino-1, 8-naphthyridine Dimer (2020)
  • Author(s): Bimolendu Das, Kazuhiko Nakatani
  • Organizer: The 23rd SANKEN INTERNATIONAL SYMPOSIUM
  • Int'l Joint Research
• [Presentation] Controlling -1 ribosomal frameshifting by small molecules in cells (2020)
  • Author(s): Asako Murata, Saki Matsumoto, Neva Caliskan, Marina V. Rodnina, Kazuhiko Nakatani
  • Organizer: The 23rd SANKEN INTERNATIONAL SYMPOSIUM
  • Int'l Joint Research
• [Presentation] Structural analysis and functionalization of naphthyridine dimer derivatives bound to CGG repeats (2020)
  • Author(s): Takeshi Yamada, Shuhei Sakurabayashi, Kyoko Huruitab, Chojiro Kojima, Kazuhiko Nakatani
  • Organizer: The 23rd SANKEN INTERNATIONAL SYMPOSIUM
  • Int'l Joint Research
• [Presentation] Small Molecules targeting Repeat Sequences causing Neurological Disorders (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: IUPAC International Symposium on Bioorganic Chemistry (ISBOC-12)
  • Int'l Joint Research / Invited
• [Presentation] Small Molecules targeting Repeat Sequences causing Neurological Disorders (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: 10th ISAJ Annual Symposium 2019
  • Invited
• [Presentation] 核酸標的低分子創薬に向けて : 何から始める? 何が足りない? (2019)
  • Author(s): 中谷 和彦
  • Organizer: 第37回メディシナルケミストリーシンポジウム
  • Invited
• [Presentation] Small Molecules targeting Repeat Sequences causing Neurological Disorders (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: Symposium on Nucleic Acids Chemistry
  • Int'l Joint Research / Invited
• [Presentation] Computer-aided molecular design for DNA bulge recognition ligands by order-N elongation method (2019)
  • Author(s): Yuuichi Orimoto, Anna Pomogaeva, Keisuke Hisama, Ayaka Yano, Koichi Miyagawa, Kazuhiko Nakatani, Yuriko Aoki
  • Organizer: 第46回国際核酸化学シンポジウム日本核酸化学会第3回年会(ISNAC2019)
  • Int'l Joint Research
• [Presentation] Novel Design Strategy of DNA-Artificial Nucleic Acid Chimeras Toward Enhancement of RNase H Cleavage Activities (2019)
  • Author(s): Masahito Inagaki, Akira Yano, Seiya Ishizawa, Masaki Nishijima, Yasuyuki Arakil, Satoru Ishibashi, Asako Yamayoshi, Kazuhiko Nakatani, Takanori Yokota, Takehiko Wada
  • Organizer: 第46回国際核酸化学シンポジウム日本核酸化学会第3回年会(ISNAC2019)
  • Int'l Joint Research
• [Presentation] DNAミスマッチ部位に結合する小分子に関する分子動力学および量子化学計算による研究 (2019)
  • Author(s): 宮川 晃一, 庄司 光男, 中谷 和彦, 重田 育照, 山口 兆
  • Organizer: 第13回分子科学討論会
• [Presentation] Dimeric Form of 1, 3-diaminoisoquinoline Derivatives Improved Alternative Splicing in Myotonic Dystrophy Type I Mouse Model (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: Asian 3 Roundtable on Nucleic Acids (A3RONA 2019)
  • Int'l Joint Research / Invited
• [Presentation] Small Molecules targeting Repeat Sequences causing Neurological Disorders (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: 7th Symposium RIKEN - Max Planck Joint Research Center for Systems Chemical Biology
  • Int'l Joint Research / Invited
• [Presentation] ANP77 : A Three-carbon Atom Linked 2-Amino-1, 8-naphthyridine Dimer that Recognizes Cytosine Rich Bulge-mismatched Sequences of Duplex DNA and RNA (2019)
  • Author(s): Bimolendu Das, Kazuhiko Nakatani
  • Organizer: 27th International Society of Heterocyclic Chemistry Congress
  • Int'l Joint Research
• [Presentation] Modulation of ribozyme activity by conformational changes induced by a synthetic RNA binding molecule (2019)
  • Author(s): Chikara Dohno, Maki Kimura, Kazuhiko Nakatani
  • Organizer: 27th International Society of Heterocyclic Chemistry Congress
  • Int'l Joint Research
• [Presentation] RNA binding small molecule that mitigates disease phenotype in spinocerebellar ataxia type 31 (2019)
  • Author(s): Tomonori Shibata, Konami Nagano, Morio Ueyama, Yoshitaka Nagai, Kinya Ishikawa, Gota Kawai, and Kazuhiko Nakatani
  • Organizer: The Commemorative International Symposium of the Japan Society of Nucleic Acids Chemistry (CISNAC 2019)
  • Int'l Joint Research
• [Presentation] Kinetic analysis of the inhibitory effect of BzDANP on Dicer cleavage of pre-miR-136 (2019)
  • Author(s): Asako Murata, Takahiro Otabe, Konami Nagano, Gota Kawai, Ayako Sugai, Kazuhiko Nakatani
  • Organizer: The Commemorative International Symposium of the Japan Society of Nucleic Acids Chemistry (CISNAC 2019)
  • Int'l Joint Research
• [Presentation] Regulation of RNA cleaving activity of hammerhead ribozyme by a synthetic RNA binding molecule (2019)
  • Author(s): Chikara Dohno, Maki Kimura, Kazuhiko Nakatani
  • Organizer: The Commemorative International Symposium of the Japan Society of Nucleic Acids Chemistry (CISNAC 2019)
  • Int'l Joint Research
• [Presentation] The methylation effect for binding of dimeric 2-amino-1, 8-naphthyridine derivatives to CGG/CGG triad in DNA (2019)
  • Author(s): Takeshi Yamada, Shuhei Sakurabayashi, Kazuhiko Nakatani
  • Organizer: The Commemorative International Symposium of the Japan Society of Nucleic Acids Chemistry (CISNAC 2019)
  • Int'l Joint Research
• [Presentation] Small Molecules targeting Repeat Sequences causing Neurological Disorders (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: The Commemorative International Symposium of the Japan Society of Nucleic Acids Chemistry (CISNAC 2019)
  • Int'l Joint Research / Invited
• [Presentation] 核酸標的低分子創薬を支えるゲノム化学 (2019)
  • Author(s): 中谷 和彦
  • Organizer: 2019 前期 有機合成化学講習会
  • Invited
• [Presentation] Structural insights into synthetic ligands targeting A-A pairs in disease-related CAG RNA repeats (2019)
  • Author(s): Sanjukta Mukherjee, Leszek Blaszczyk, Wojciech Rypniewski, Christoph Falschlunger, Ronald Micura, Asako Murata, Chikara Dohno, Kazuhiko Nakatani, Agnieszka Kiliszek
  • Organizer: The 24th Annual Meeting of the RNA Society (RNA 2019)
  • Int'l Joint Research
• [Presentation] Exploration of RNA sequences in pre-miRNA affecting the efficiency of Dicer cleavage reactions by the small molecules binding (2019)
  • Author(s): Yusuke Takashima, Asako Murata, Kazuhiko Nakatani
  • Organizer: The 24th Annual Meeting of the RNA Society (RNA 2019)
  • Int'l Joint Research
• [Presentation] XGGリピートDNA中のピリミジン塩基を選択的に酸化するリガンドの開発 (2019)
  • Author(s): 八木 勇樹、山田 剛史、中谷 和彦
  • Organizer: 日本ケミカルバイオロジー学会 第14回年会
• [Presentation] GGCCCCリピート結合分子N, N’-ジヘテロアリールグアニジン誘導体の合成及び結合評価 (2019)
  • Author(s): 村上 英太郎、柴田 知範、中谷 和彦
  • Organizer: 日本ケミカルバイオロジー学会 第14回年会
• [Presentation] 脊髄小脳変性症31型を標的とするRNA結合性低分子 (2019)
  • Author(s): 柴田 知範、長野 来南、上山 盛夫、永井 義隆、石川 欽也、河合 剛太、中谷 和彦
  • Organizer: 日本ケミカルバイオロジー学会 第14回年会
• [Presentation] Small Molecules targeting Trinucleotide Repeat Sequences causing Neurological Disorders (2019)
  • Author(s): Kazuhiko Nakatani
  • Organizer: Trends in Nucleic Acid
  • Int'l Joint Research / Invited
• [Remarks]
  • URL: http://www.sanken.osaka-u.ac.jp/labs/rbc/