Sequence specific double-strand DNA cleavage by an optical active dinuclear iron complex and application for artificial restriction enzyme

光学活性双核铁复合物对序列特异性双链 DNA 的切割及其人工限制性内切酶的应用

• 批准号: 14572034
• 负责人: KUROSAKI Hiromasa
• 金额: $ 2.18万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572034/
• 关键词: iron complex; DNA cleavage; optical active; electronic spectra; CD spectra; 過酸化水素; 酸化反応; ESR; CDスペクトル

1-1)In my approach to create metal complexes that are capable of cleaving double-strand DNA simultaneously and sequence-selectively (like bleomycin, which is an antitumor antibiotic which causes single and double strand cleavage of DNA at 5'-GC-3' or 5'-GT-3' sites), I prepared an optical active pentadentate ligand, N,N,N'-tri(2-pyridylmethyl)-2-S-(aminomethyl)piperidine = S-P3pipda obtained from the reaction of S-2-aminopiperidine with 3 equimolar amounts of 2-chloromethylpyridine, and investigated the X-ray crystal structures of copper(II) and nickel(II) complexes of S-P3pipda, [Cu(S-P3pipda)](ClO_4)_2 and [Ni(S-P3pipda)(OH_2)](ClO_4)_2 Complexes, [Cu(S-P3pipda)](ClO_4)_2 and [Ni(S-P3pipda)(OH_2)](ClO_4)_2, were prepared from the reaction of an equimolar amount of Cu(ClO_4)_2・6H_2O or Ni(SO_4)_2・6H_2O (followed by addition of LiClO_4) with the ligand S-P3pipda in methanol under argon.The X-ray crystal structure of [Cu(S-P3pida)]^<2+> shows an intermediate structure between trigonal bipyramid and square pyramid in which equatrial positions are occupied by two tertiary amines (N2 and N3) and two pyridine nitrogens (N1 and N4) and axial position by a pyridine nitrogen (N5). Perchlorate ion coordinates weakly : Cu-O is 3.55 Å.1-2)We synthesized 1,8-(N-anthranylmethyl)-bis(N',N'-dipyridylmethyl-R,R-cyclohexane-1,2-diamine), 1, and cleaving activity of 1 to DNA was evaluated in the presence of sodium ascorbate under air. A novel Fe complex 1 and sodium ascorbate system cleaved A-T rich region of DNA.2)Moreover, I have synthesized two new dinuclear platinum(II) complexes, 1,3-and 1,4-(benzenedimethanethiolate-S)di(2,2',2"-terpyridine)platinum(II) chloride tetrahydrate (2 and 3), were synthesized in order to investigate the binding of platinum(II) complexes with calf thymus DNA, which was examined by UV and CD spectroscopies. Complex 2 interacted strongly with DNA by intercalation compared to 3.

1-1) 在我创建能够同时和序列选择性地切割双链 DNA 的金属复合物的方法中（如博来霉素，它是一种抗肿瘤抗生素，会导致 5'-GC-3' 或 5'-GT-3' 位点处的 DNA 单链和双链切割），我制备了一种光学活性五齿配体， 由S-2-氨基哌啶与3等摩尔量的2-氯甲基吡啶反应得到N,N,N'-三(2-吡啶基甲基)-2-S-(氨甲基)哌啶=S-P3pipda，并研究了S-P3pipda铜(II)和镍(II)络合物的X射线晶体结构， [Cu(S-P3pipda)](ClO_4)_2 和 [Ni(S-P3pipda)(OH_2)](ClO_4)_2 配合物，[Cu(S-P3pipda)](ClO_4)_2 和 [Ni(S-P3pipda)(OH_2)](ClO_4)_2 是由等摩尔量的反应制备的 Cu(ClO_4)_2·6H_2O 或 Ni(SO_4)_2·6H_2O（随后添加 LiClO_4）与配体 S-P3pipda 在甲醇中于氩气下反应。 [Cu(S-P3pida)]^<2+> 的 X 射线晶体结构显示出介于三角双锥和四方锥之间的中间结构，其中等方 位置被两个叔胺（N2和N3）和两个吡啶氮（N1和N4）占据，轴向位置被吡啶氮（N5）占据。高氯酸根离子配位较弱：Cu-O为3.55 Å.1-2)我们合成了1,8-(N-蒽基甲基)-双(N',N'-二吡啶基甲基-R,R-环己烷-1,2-二胺),1，并在空气中抗坏血酸钠存在下评估了1对DNA的切割活性。新型 Fe 配合物 1 和抗坏血酸钠系统可裂解 DNA 富含 A-T 的区域。2)此外，我合成了两种新的双核铂 (II) 配合物，1,3-和 1,4-(苯二甲硫醇-S)二(2,2',2"-三联吡啶)氯化铂(II)四水合物（2 和 3），以研究 铂 (II) 络合物与小牛胸腺 DNA 的结合，通过 UV 和 CD 光谱进行检查。与 3 相比，复合物 2 通过嵌入与 DNA 发生强烈相互作用。

项目成果

Masafumi Goto, Yasuhiko Ohse, Sayuri Koga, Yuka Kudo, Rinko Kukihara, Hiromasa Kurosaki: "Preparations of Low-Spin Tetra Cyano Iron Complexes of N-Benzyl-1,2-ethanediamine and Its Oxidative Dehydrogenation : Isolation of the Intermediate with Monoimine Li

Masafumi Goto、Yasuhiko Ohse、Sayuri Koga、Yuka Kudo、Rinko Kukihara、Hiromasa Kurosaki：“N-苄基-1,2-乙二胺的低自旋四氰基铁配合物的制备及其氧化脱氢：用单亚胺锂分离中间体

Hiromasa Kurosaki, Toshiaki Tawada, Satomi Kawasoe, Yoko Ohashi, Masafumi Goto: "Crystal Sturucture of Chloro{N-mercaptoethyl[bis(6-methyl-2-pyridyl-methyl)amine]}zinc(II) Complex"Anal. Sci.. 19. x27-x28 (2003)

Hiromasa Kurosaki、Toshiaki Tawada、Satomi Kawasoe、Yoko Ohashi、Masafumi Goto：“氯{N-巯基乙基[双(6-甲基-2-吡啶基-甲基)胺]}锌(II)络合物的晶体结构”分析。

Masafumi Goto, Yuuichiro Yamamoto, Masamisu Sumimoto, Nobuhiro Tsuruda, Hiromasa Kurosaki: "Conformation of 2-Aminomethylpyrrolidine and 2-Aminomethylpiperidine in Ternary Platinum(II) Complexes : Regulation of Conformation of the Diamine by the Coexistin

Masafumi Goto、Yuuichiro Yamamoto、Masamisu Sumimoto、Nobuhiro Tsuruda、Hiromasa Kurosaki：“三元铂 (II) 配合物中 2-氨基甲基吡咯烷和 2-氨基甲基哌啶的构象：共存蛋白对二胺构象的调节

Masafumi Goto: "DNA Cleavage by Pentadentate Iron(II) Complexes Containing Fluoro-Substituted Phenyl Groups"Bioorg.Med.Chem.Lett.. 12. 201-203 (2002)

Masafumi Goto：“含有氟取代苯基的五齿铁 (II) 配合物对 DNA 的切割”Bioorg.Med.Chem.Lett.. 12. 201-203 (2002)

Masafumi Goto: "Interaction of Binuclear Xylylthiolato(2,2',2"-terpyridine)platinum(II) Complexes with DNA"Bioorg.Med.Chem.Lett.. 13. 825-828 (2003)

Masafumi Goto：“双核二甲苯硫基(2,2,2”-三联吡啶)铂(II)复合物与DNA的相互作用”Bioorg.Med.Chem.Lett.. 13. 825-828 (2003)