研究者詳細 - 阿部　洋

論文 - 阿部　洋

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Synthesis of hydrophobic-tagged 2′-deoxy-modified cap analogs and its effect on mRNA translation

Zheyu Meng, Yuko Nakashima, Masahito Inagaki, Zhenmin Li, Susit Acharyya, Fumitaka Hashiya, Naoko Abe, Yasuaki Kimura, Hiroshi Abe

BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN 98 巻 ( 2 ) 2025年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：OXFORD UNIV PRESS

Messenger RNA (mRNA) has been implemented as COVID-19 vaccines. Following their success, the application of mRNA in cancer vaccines and protein replacement therapies is highly anticipated. The translational efficiency of mRNA largely depends on the cap structure located at its 5 ' end. In particular, the Cap-1 and Cap-2 structures are known to exhibit higher protein synthesis levels than the Cap-0 structure. Previously, we developed a method called the PureCap technique, which utilizes reversed-phase high-performance liquid chromatography (RP-HPLC) to purify highly capped mRNA with high precision, enabling accurate evaluation of translational activity. In this study, we synthesized cap analogs incorporating deoxyribose, and evaluated their mRNA synthesis efficiency and translational activity. Furthermore, by optimizing the transcription reaction conditions using the synthesized DNA-modified Cap series, we successfully improved the capping efficiency and mRNA yield. The deoxyadenosine-modified cap analog synthesized using the PureCap technique demonstrated 3.2-fold higher translational activity than the standard Cap-1 structure. Additionally, when using tetranucleotide cap analogs, consistent translational activity was observed, regardless of base sequence differences. This confirmed the general applicability of DNA-modified mRNA for translational activity.

DOI： 10.1093/bulcsj/uoaf006

Internal cap-initiated translation for efficient protein production from circular mRNA

Fukuchi K., Nakashima Y., Abe N., Kimura S., Hashiya F., Shichino Y., Liu Y., Ogisu R., Sugiyama S., Kawaguchi D., Inagaki M., Meng Z., Kajihara S., Tada M., Uchida S., Li T.T., Maity R., Kawasaki T., Kimura Y., Iwasaki S., Abe H.

Nature Biotechnology 2025年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nature Biotechnology

Circular mRNA faces challenges in enhancing its translation potential as an RNA therapeutic. Here we introduce two molecular designs that bolster circular mRNA translation through an internal cap-initiated mechanism. The first consists of a circular mRNA with a covalently attached N7-methylguanosine (m7G) cap through a branching structure (cap-circ mRNA). This modification allows circular mRNA to recruit translation machinery and produce proteins more efficiently than internal ribosome entry site (IRES)-containing circular mRNAs. Combining with an N1-methylpseudouridine (m1Ψ) modification, cap-circ mRNA exhibits a lower acute immunostimulatory effect, maintaining high translation in mice. The second design features the non-covalent attachment of an m7G cap to a circular mRNA through hybridization with an m7G cap-containing oligonucleotide, enhancing translation by more than 50-fold. This setup allows circular mRNAs to synthesize reporter proteins upon hybridizing with capped mRNAs or long non-coding RNAs and to undergo rolling circle-type translation. These advancements broaden the therapeutic applications of circular mRNAs by minimizing their molecular size, elevating translation efficiency and facilitating cell-type-selective translation.

DOI： 10.1038/s41587-025-02561-8

Covalently or non-covalently attached m<SUP>7</SUP>G cap enhances protein production from circular mRNA 招待有り査読有り国際誌

Hiroshi Abe, Naoko Abe

NATURE BIOTECHNOLOGY 2025年2月

Concise Affinity-Based Purification of Ligated mRNA for Structure-Activity Relationship Studies of Nucleosugar Modification Patterns 査読有り国際誌

Yamada H., Iwai H., Hashiya F., Kimura Y., Abe H., Yamamoto J.

ChemBioChem 26 巻 ( 2 ) 頁： e202400711 2025年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemBioChem

Position-specific nucleoside sugar modifications have been shown to improve the translational activity and stability of chemically synthesized mRNA. For pharmaceutical applications of chemically modified mRNAs, a rapid purification methodology is imperative to identify the optimal modification pattern. However, while the chemical synthesis of mRNAs can be accomplished by splint ligation of oligonucleotide fragments, the current purification method for ligated mRNAs based on denaturing polyacrylamide gel electrophoresis tends to be time consuming. In this study, we developed a two-step affinity purification method for rapid sample preparation. In this method, ligated mRNA is captured by oligo dT magnetic beads and streptavidin magnetic beads with 3’-biotinylated oligo DNA, which are complementary to the 3’-poly(A) and 5’ terminal sequences of the target mRNA, respectively. Therefore, the target mRNA can be isolated from a complex mixture of splint ligations. Using this method, six sugar-modified mRNAs were simultaneously purified, and the translational activities of these mRNAs were evaluated immediately after purification. The results demonstrate that this methodology is suitable for the rapid preparation of various chemically synthesized mRNAs to identify their optimal modification patterns.

DOI： 10.1002/cbic.202400711

Synthesis of Novel Cap Analogs Bearing Hydrophobic Purification Tag Toward Application for Preparation of Fully Capped Messenger RNA

Inagaki M., Abe H.

Yuki Gosei Kagaku Kyokaishi/Journal of Synthetic Organic Chemistry 83 巻 ( 1 ) 頁： 2 - 14 2025年1月

出版者・発行元：Yuki Gosei Kagaku Kyokaishi/Journal of Synthetic Organic Chemistry

Messenger RNA (mRNA) is gaining attention as a new modality of oligonucleotide therapeutics. We have reported a new mRNA synthesis method, named the PureCap method, by in vitro transcription, which enables the production of completely capped mRNA. For this purpose, we designed and synthesized new cap analogs bearing hydrophobic o–nitrobenzyl groups, which function as purification tags for mRNA with a 5́–cap structure by reverse–phase high–performance liquid chromatography. In this paper, we describe the synthesis of new cap analogs, named PureCap analogs, and its application for completely capped mRNA synthesis.

DOI： 10.5059/yukigoseikyokaishi.83.2

Synthesis of 2′-formamidonucleoside phosphoramidites for suppressing the seed-based off-target effects of siRNAs 査読有り国際誌 Open Access

Nomura K., An S., Kobayashi Y., Kondo J., Shi T., Murase H., Nakamoto K., Kimura Y., Abe N., Ui-Tei K., Abe H.

Nucleic Acids Research 52 巻 ( 18 ) 頁： 10754 - 10774 2024年9月

担当区分：最終著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic Acids Research

In this study, we report the synthesis of 2′-formamidonucleoside phosphoramidite derivatives and their incorporation into siRNA strands to reduce seed-based off-target effects of small interfering RNAs (siRNAs). Formamido derivatives of all four nucleosides (A, G, C and U) were synthesized in 5-11 steps from commercial compounds. Introducing these derivatives into double-stranded RNA slightly reduced its thermodynamic stability, but X-ray crystallography and CD spectrum analysis confirmed that the RNA maintained its natural A-form structure. Although the introduction of the 2′-formamidonucleoside derivative at the 2nd position in the guide strand of the siRNA led to a slight decrease in the on-target RNAi activity, the siRNAs with different sequences incorporating 2′-formamidonucleoside with four kinds of nucleobases into any position other than 2nd position in the seed region revealed a significant suppression of off-target activity while maintaining on-target RNAi activity. This indicates that 2′-formamidonucleosides represent a promising approach for mitigating off-target effects in siRNA therapeutics.

DOI： 10.1093/nar/gkae741

2′-β-Methylselenyl nucleos(t)ide analogs as reverse transcriptase inhibitors against diverse HIV mutants 査読有り国際誌 Open Access

Yoshida Y., Niimi Y., Fushihara D., Katakura H., Fukui R., Murase H., Tomoike F., Hashiya F., Murakami T., Kodama E.N., Suzuki T., Yasukawa K., Kimura Y., Abe H.

Bioorganic and Medicinal Chemistry 110 巻頁： 117813 - 117813 2024年8月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry

Nucleoside reverse transcriptase inhibitors (NRTIs) have been extensively studied as drugs targeting HIV RT. However, the practice or use of approved NRTIs lacking the 3ʹ-hydroxy group often promotes frequent HIV mutations and generates drug-resistance. Here, we describe a novel NRTI with 2′-β-methylselenyl modification. We found that this modification inhibited the DNA elongation reaction by HIV-1 RT despite having a 3′-hydroxy group. Moreover, the conformation of this nucleoside analog is controlled at C3′-endo, a conformation that resists excision from the elongating DNA by HIV RT. Accordingly, the designed analogs exhibited activity against both wild-type HIV and multidrug-resistant HIV mutants.

DOI： 10.1016/j.bmc.2024.117813

Intracellular Delivery of Antisense Oligonucleotides by Tri-Branched Cyclic Disulfide Units 査読有り国際誌 Open Access

Lyu F., Hakariya H., Hiraoka H., Li Z., Matsubara N., Soo Y., Hashiya F., Abe N., Shu Z., Nakamoto K., Kimura Y., Abe H.

ChemMedChem 19 巻 ( 20 ) 頁： e202400472 2024年7月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemMedChem

Therapeutic oligonucleotides, such as antisense DNA, show promise in treating previously untreatable diseases. However, their applications are still hindered by the poor membrane permeability of naked oligonucleotides. Therefore, it is necessary to develop efficient methods for intracellular oligonucleotide delivery. Previously, our group successfully developed disulfide-based Membrane Permeable Oligonucleotides (MPON), which achieved enhanced cellular uptake and gene silencing effects through an endocytosis-free uptake mechanism. Herein, we report a new molecular design for the next generation of MPON, called trimer MPON. The trimer MPON consists of a tri-branched backbone, three α-lipoic acid units, and a spacer linker between the oligonucleotides and tri-branched cyclic disulfide unit. We describe the design, synthesis, and functional evaluation of the trimer MPON, offering new insights into the molecular design for efficient oligonucleotide delivery.

DOI： 10.1002/cmdc.202400472

Development and Comparison of 4-Thiouridine to Cytidine Base Conversion Reaction 招待有り査読有り国際誌 Open Access

Sana Ohashi, Mayu Nakamura, Susit Acharyya, Masahito Inagaki, Naoko Abe, Yasuaki Kimura, Fumitaka Hashiya, Hiroshi Abe

ACS Omegam 9 巻 ( 8 ) 頁： 9300 - 9308 2024年2月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACS Omega

To study transcriptome dynamics without harming cells, it is essential to convert chemical bases. 4-Thiouridine (4sU) is a biocompatible uridine analogue that can be converted into a cytidine analogue. Although several reactions can convert 4sU into a cytidine analogue, few studies have compared the features of these reactions. In this study, we performed three reported base conversion reactions, including osmium tetroxide, iodoacetamide, and sodium periodate treatment, as well as a new reaction using 2,4-dinitrofluorobenzene. We compared the reaction time, conversion efficacy, and effects on reverse transcription. These reactions successfully converted 4sU into a cytidine analogue quantitatively using trinucleotides. However, the conversion efficacy and effect on reverse transcription vary depending on the reaction with the RNA transcript. OsO4 treatment followed by NH4Cl treatment showed the best base-conversion efficiency. Nevertheless, each reaction has its own advantages and disadvantages as a tool for studying the transcriptome. Therefore, it is crucial to select the appropriate reaction for the target of interest.

DOI： 10.1021/acsomega.3c08516

Development of PCR primers enabling the design of flexible sticky ends for efficient concatenation of long DNA fragments 査読有り国際誌 Open Access

Nomura K., Onda K., Murase H., Hashiya F., Ono Y., Terai G., Oka N., Asai K., Suzuki D., Takahashi N., Hiraoka H., Inagaki M., Kimura Y., Shimizu Y., Abe N., Abe H.

RSC Chemical Biology 5 巻 ( 4 ) 頁： 360 - 371 2024年2月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：RSC Chemical Biology

We developed chemically modified PCR primers that allow the design of flexible sticky ends by introducing a photo-cleavable group at the phosphate moiety. Nucleic acid derivatives containing o-nitrobenzyl photo-cleavable groups with a tert-butyl group at the benzyl position were stable during strong base treatment for oligonucleotide synthesis and thermal cycling in PCR reactions. PCR using primers incorporating these nucleic acid derivatives confirmed that chain extension reactions completely stopped at position 1 before and after the site of the photo-cleavable group was introduced. DNA fragments of 2 and 3 kbp, with sticky ends of 50 bases, were successfully concatenated with a high yield of 77%. A plasmid was constructed using this method. Finally, we applied this approach to construct a 48.5 kbp lambda phage DNA, which is difficult to achieve using restriction enzyme-based methods. After 7 days, we were able to confirm the generation of DNA of the desired length. Although the efficiency is yet to be improved, the chemically modified PCR primer offers potential to complement enzymatic methods and serve as a DNA concatenation technique.

DOI： 10.1039/d3cb00212h

Topological capture of mRNA for silencing gene expression 査読有り国際誌

Lyu F., Tomita T., Abe N., Hiraoka H., Hashiya F., Nakashima Y., Kajihara S., Tomoike F., Shu Z., Onizuka K., Kimura Y., Abe H.

Chemical Communications 59 巻 ( 77 ) 頁： 11564 - 11567 2023年9月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

We describe herein topological mRNA capture using branched oligodeoxynucleotides (ODNs) with multiple reactive functional groups. These fragmented ODNs efficiently formed topological complexes on template mRNA in vitro. In cell-based experiments targeting AcGFP mRNA, the bifurcated reactive ODNs showed a much larger gene silencing effect than the corresponding natural antisense ODN.

DOI： 10.1039/d2cc06189a

The Effect of γ Phosphate Modified Deoxynucleotide Substrates on PCR Activity and Fidelity 査読有り国際誌 Open Access

Hashiya, F; Murase, H; Chandela, A; Hiraoka, H; Inagaki, M; Nakashima, Y; Abe, N; Nakamura, M; Terai, G; Kimura, Y; Ando, K; Oka, N; Asai, K; Abe, H

CHEMBIOCHEM 24 巻 ( 14 ) 頁： e202200572 2023年5月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemBioChem

Controlling PCR fidelity is an important issue for molecular biology and high-fidelity PCR is essential for gene cloning. In general, fidelity control is achieved by protein engineering of polymerases. In contrast, only a few studies have reported controlling fidelity using chemically modified nucleotide substrates. In this report, we synthesized nucleotide substrates possessing a modification on Pγ and evaluated the effect of this modification on PCR fidelity. One of the substrates, nucleotide tetraphosphate, caused a modest decrease in Taq DNA polymerase activity and the effect on PCR fidelity was dependent on the type of mutation. The use of deoxyadenosine tetraphosphate enhanced the A : T→G : C mutation dramatically, which is common when using Taq polymerase. Conversely, deoxyguanosine tetraphosphate (dG4P) suppressed this mutation but increased the G : C→A : T mutation during PCR. Using an excess amount of dG4P suppressed both mutations successfully and total fidelity was improved.

DOI： 10.1002/cbic.202200572

Synthesis of nucleoside oligophosphates by electrophilic activation of phosphorothioate 査読有り国際誌

Hasegawa, S; Inagaki, M; Kato, S; Li, ZM; Kimura, Y; Abe, H

ORGANIC & BIOMOLECULAR CHEMISTRY 21 巻 ( 19 ) 頁： 3997 - 4001 2023年5月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Organic and Biomolecular Chemistry

We herein report a new synthetic method for nucleoside oligophosphates based on electrophilic activation of 5′-phosphorothioate nucleotides. The treatment of phosphorothioate with 2,4-dinitrochlorobenzene (DNCB) efficiently afforded the key activated species, electrophilic thioester nucleotides (EPT-Ns), which were coupled with various phosphate reagents to afford the target nucleoside oligophosphates, including an mRNA cap analog.

DOI： 10.1039/d2ob02260e

Cap analogs with a hydrophobic photocleavable tag enable facile purification of fully capped mRNA with various cap structures 査読有り国際誌 Open Access

Inagaki, M; Abe, N; Li, ZM; Nakashima, Y; Acharyya, S; Ogawa, K; Kawaguchi, D; Hiraoka, H; Banno, A; Meng, ZY; Tada, M; Ishida, T; Lyu, P; Kokubo, K; Murase, H; Hashiya, F; Kimura, Y; Uchida, S; Abe, H

NATURE COMMUNICATIONS 14 巻 ( 1 ) 頁： 2657 - 2657 2023年5月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nature Communications

Starting with the clinical application of two vaccines in 2020, mRNA therapeutics are currently being investigated for a variety of applications. Removing immunogenic uncapped mRNA from transcribed mRNA is critical in mRNA research and clinical applications. Commonly used capping methods provide maximum capping efficiency of around 80–90% for widely used Cap-0- and Cap-1-type mRNAs. However, uncapped and capped mRNA possesses almost identical physicochemical properties, posing challenges to their physical separation. In this work, we develop hydrophobic photocaged tag-modified cap analogs, which separate capped mRNA from uncapped mRNA by reversed-phase high-performance liquid chromatography. Subsequent photo-irradiation recovers footprint-free native capped mRNA. This approach provides 100% capping efficiency even in Cap-2-type mRNA with versatility applicable to 650 nt and 4,247 nt mRNA. We find that the Cap-2-type mRNA shows up to 3- to 4-fold higher translation activity in cultured cells and animals than the Cap-1-type mRNA prepared by the standard capping method.

DOI： 10.1038/s41467-023-38244-8

Creation of single molecular conjugates of metal-organic cages and DNA 査読有り国際誌 Open Access

Nakajo, T; Kusaka, S; Hiraoka, H; Nomura, K; Matsubara, N; Baba, R; Yoshida, Y; Nakamoto, K; Honma, M; Iguchi, H; Uchihashi, T; Abe, H; Matsuda, R

CHEMICAL COMMUNICATIONS 59 巻 ( 33 ) 頁： 4974 - 4977 2023年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

Here we report the development of an equimolar conjugate of a metal-organic cage (MOC) and DNA (MOC-DNA). Several MOC-DNA conjugates were assembled into a programmed structure by coordinating with a template DNA having a complementary base sequence. Moreover, conjugation with the MOC drastically enhanced the permeability of DNA through the lipid bilayer, presenting great potential as a drug delivery system.

DOI： 10.1039/d3cc00460k

Use of Iontophoresis Technology for Transdermal Delivery of a Minimal mRNA Vaccine as a Potential Melanoma Therapeutic 査読有り Open Access

Husseini, RA; Abe, N; Hara, T; Abe, H; Kogure, K

BIOLOGICAL & PHARMACEUTICAL BULLETIN 46 巻 ( 2 ) 頁： 301 - 308 2023年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Biological and Pharmaceutical Bulletin

mRNA vaccines have attracted considerable attention as a result of the 2019 coronavirus pandemic; however, challenges remain regarding use of mRNA vaccines, including insufficient delivery owing to the high molecular weights and high negative charges associated with mRNA. These characteristics of mRNA vaccines impair intracellular uptake and subsequent protein translation. In the current study, we prepared a minimal mRNA vaccine encoding a tumor associated antigen human gp10025-33 peptide (KVPRNQDWL), as a potential treatment for melanoma. Minimal mRNA vaccines have recently shown promise at improving the translational process, and can be prepared via a simple production method. Moreover, we previously reported the successful use of iontophoresis (IP) technology in the delivery of hydrophilic macromolecules into skin layers, as well as intracellular delivery of small interfering RNA (siRNA). We hypothesized that combining IP technology with a newly synthesized minimal mRNA vaccine can improve both transdermal and intracellular delivery of mRNA. Following IP-induced delivery of a mRNA vaccine, an immune response is elicited resulting in activation of skin resident immune cells. As expected, combining both technologies led to potent stimulation of the immune system, which was observed via potent tumor inhibition in mice bearing melanoma. Additionally, there was an elevation in mRNA expression levels of various cytokines, mainly interferon (IFN)-γ, as well as infiltration of cytotoxic CD8_ T cells in the tumor tissue, which are responsible for tumor clearance. This is the first report demonstrating the application of IP for delivery of a minimal mRNA vaccine as a potential melanoma therapeutic.

DOI： 10.1248/bpb.b22-00746

Chemical synthesis of super messenger RNA medicine 査読有り

Tada Mizuki, Naoko Abe, Hiroshi Abe

Drug Delivery System 38 巻 ( 1 ) 頁： 15 - 23 2023年

Chemical synthesis of super messenger RNA medicine 査読有り Open Access

Tada M., Abe N., Abe H.

Drug Delivery System 38 巻 ( 1 ) 頁： 15 - 23 2023年

担当区分：最終著者,　責任著者記述言語：日本語掲載種別：研究論文（研究会，シンポジウム資料等）出版者・発行元：Drug Delivery System

Messenger RNA（mRNA）medicine was urgently approved in 2020 as a vaccine for COVID-19 . However, current mRNA therapeutics are not fully established, with challenges remaining in translation efficiency and drug delivery system. Therefore, further research is needed to adapt mRNA therapeutics to other diseases. Furthermore, the preparation of mRNA drugs is time-consuming and costly because of the biological methods used. Our laboratory has been working on chemical methods to solve these issues. In this paper, we introduce chemical modifications and novel capping reactions as a method to improve the translation efficiency of mRNA and the introduction of disulfide modification to oligonucleotide therapeutics as an effort on the drug delivery system.

DOI： 10.2745/dds.38.15

mRNAの完全化学合成

小川和哉, 阿部洋

2022年10月

A 2′-modified uridine analog, 2'-<i>O</i>-(methylthiomethoxy)methyl uridine, for siRNA applications 査読有り国際誌 Open Access

Lyu, F; An, S; Kobayashi, Y; Nomura, K; Baba, R; Abe, N; Hiraoka, H; Hashiya, F; Shu, ZM; Ui-Tei, K; Kimura, Y; Abe, H

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 74 巻頁： 128939 - 128939 2022年8月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

The medicinal applications of siRNAs have been intensively examined but are still hindered by their low molecular stability under biological conditions and off-target effects, etc. The introduction of chemical modifications to the nucleoside is a promising strategy for solving these limitations. Herein, we describe the development of a new uridine analog, U*, that has a (methylthiomethoxy)methoxy group at the 2′ position. The phosphoramidite reagent corresponding to U* was easily synthesized and the RNA oligonucleotides containing U* were stably prepared using a standard protocol for oligonucleotide synthesis. The introduction of U* into the siRNA resulted in positive or negative effects on the targeted gene silencing in a position-dependent manner, and the positive effects were attributed to the improved stability under biological conditions. The thermodynamic analysis of the U*-modified RNAs revealed a slight destabilization of the dsRNA, based depending on which U was strategically utilized to restrain the off-target effects of the siRNA. This study describes a rare example of nucleoside analogs with a large substitution at the 2′-position in the context of an siRNA application and is informative for the development of other analogs to further improve the molecular properties of siRNAs for medicinal applications.

DOI： 10.1016/j.bmcl.2022.128939

翻訳反応を最大化するための合成 mRNA の分子設計査読有り

稲垣雅仁, 多田瑞紀, 阿部洋

37 巻 ( 3 ) 頁： 196 - 208 2022年7月

Complete Chemical Synthesis of Minimal Messenger RNA by Efficient Chemical Capping Reaction 査読有り国際誌

Abe, N; Imaeda, A; Inagaki, M; Li, ZM; Kawaguchi, D; Onda, K; Nakashima, Y; Uchida, S; Hashiya, F; Kimura, Y; Abe, H

ACS CHEMICAL BIOLOGY 17 巻 ( 6 ) 頁： 1308 - 1314 2022年5月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACS Chemical Biology

Site-specific chemical modification of mRNA can improve its translational efficiency and stability. For this purpose, it is desirable to develop a complete chemical synthesis method for chemically modified mRNA. The key is a chemical reaction that introduces a cap structure into the chemically synthesized RNA. In this study, we developed a fast and quantitative chemical capping reaction between 5′-phosphorylated RNA and N7-methylated GDP imidazolide in the presence of 1-methylimidazole in the organic solvent dimethyl sulfoxide. It enabled quantitative preparation of capping RNA within 3 h. We prepared chemically modified 107-nucleotide mRNAs, including N6-methyladenosine, insertion of non-nucleotide linkers, and 2′-O-methylated nucleotides at the 5′ end and evaluated their effects on translational activity in cultured HeLa cells. The results showed that mRNAs with non-nucleotide linkers in the untranslated regions were sufficiently tolerant to translation and that mRNAs with the Cap_2 structure had higher translational activity than those with the Cap_0 structure.

DOI： 10.1021/acschembio.1c00996

Development of Fluorophosphoramidate as a Biocompatibly Transformable Functional Group and its Application as a Phosphate Prodrug for Nucleoside Analogs 査読有り国際誌

Yoshida Yuki, Zheng Ti, Tanabe Wataru, Tomoike Fumiaki, Hashiya Fumitaka, Suzuki Tetsuro, Hirota Shuto, Saiki Yuriko, Horii Akira, Hirayama Akiyoshi, Soga Tomoyosi, Kimura Yasuaki, Abe Hiroshi

CHEMMEDCHEM 17 巻 ( 17 ) 頁： e202200188 2022年4月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemMedChem

Synthetic phosphate-derived functional groups are important for controlling the function of bioactive molecules in vivo. Herein we describe the development of a new type of biocompatible phosphate analog, a fluorophosphoramidate (FPA) functional group that has characteristic P−F and P−N bonds. We found that FPA with a primary amino group was relatively unstable in aqueous solution and was converted to a monophosphate, while FPA with a secondary amino group was stable. Furthermore, by improving the molecular design of FPA, we developed a reaction in which a secondary amino group is converted to a primary amino group in the intracellular environment and clarified that the FPA group functions as a phosphate prodrug of nucleoside. Various FPA-gemcitabine derivatives were synthesized and their toxicity to cancer cells were evaluated. One of the FPA-gemcitabine derivatives showed superior toxicity compared with gemcitabine and its ProTide prodrug, which methodology is widely used in various nucleoside analogs, including anti-cancer and anti-virus drugs.

DOI： 10.1002/cmdc.202200188

Ligandability Assessment of Human Glutathione Transferase M1-1 Using Pesticides as Chemical Probes 査読有り国際誌

Bodourian Charoutioun S., Poudel Nirmal, Papageorgiou Anastassios C., Antoniadi Mariana, Georgakis Nikolaos D., Abe Hiroshi, Labrou Nikolaos E.

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 23 巻 ( 7 ) 2022年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：International Journal of Molecular Sciences

Glutathione transferases (GSTs; EC 2.5.1.18) form a group of multifunctional enzymes that are involved in phase II of the cellular detoxification mechanism and are associated with increased susceptibility to cancer development and resistance to anticancer drugs. The present study aims to evaluate the ligandability of the human GSTM1-1 isoenzyme (hGSTM1-1) using a broad range of struc-turally diverse pesticides as probes. The results revealed that hGSTM1-1, compared to other classes of GSTs, displays limited ligandability and ligand-binding promiscuity, as revealed by kinetic inhibition studies. Among all tested pesticides, the carbamate insecticide pirimicarb was identified as the strong-est inhibitor towards hGSTM1-1. Kinetic inhibition analysis showed that pirimicarb behaved as a mixed-type inhibitor toward glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). To shine a light on the restricted hGSTM1-1 ligand-binding promiscuity, the ligand-free crystal structure of hGSTM1-1 was determined by X-ray crystallography at 1.59 Å-resolution. Comparative analysis of ligand-free structure with the available ligand-bound structures allowed for the study of the enzyme’s plasticity and the induced-fit mechanism operated by hGSTM1-1. The results revealed important structural features of the H-site that contribute to xenobiotic-ligand binding and specificity. It was concluded that hGSTM1-1 interacts preferentially with one-ring aromatic compounds that bind at a discrete site which partially overlaps with the xenobiotic substrate binding site (H-site). The results of the study form a basis for the rational design of new drugs targeting hGSTM1-1.

DOI： 10.3390/ijms23073606

Design of Synthetic mRNAs for Highly Ef ficient Translation 招待有り査読有り Open Access

Inagaki M., Tada M., Abe H.

Drug Delivery System 37 巻 ( 3 ) 頁： 196 - 208 2022年

担当区分：最終著者,　責任著者記述言語：日本語掲載種別：研究論文（研究会，シンポジウム資料等）出版者・発行元：Drug Delivery System

Recently messenger RNA （mRNA）therapeutics is received much attention as one of the vaccination therapies to compete against the coronavirus disease 2019（COVID-19）pandemic. DDS mRNA therapeutics are generally produced by in vitro transcription utilizing RNA polymerase mediated elongation. However, its purity, stability, and protein synthesis ability, are difficult to be precisely controlled, which is pointed out as drawbacks that must be overcome. To overcome these issues, the introduction of chemically modified nucleic acids is focusing attention. However, it is difficult to flexible molecular design due to the requirement of RNA polymerase recognition ability of chemically modified nucleic acids under in vitro transcription reaction. In the future, the development of a new mRNA design concept based on a flexible molecular design by the progress of chemically modified mRNA therapeutics synthesis method. Under the situation, the authors are focusing on the translation mechanism of mRNA and proposing a new mRNA molecular design to accelerate the translation reaction cycle. In this paper, we introduce an update on therapeutic mRNA design.

DOI： 10.2745/dds.37.196

Completely Chemically Synthesized Long DNA Can be Transcribed in Human Cells

Yamaoka Kazuki, Oikawa Ryota, Abe Naoko, Nakamoto Kosuke, Tomoike Fumiaki, Hashiya Fumitaka, Kimura Yasuaki, Abe Hiroshi

CHEMBIOCHEM 22 巻 ( 23 ) 頁： 3273 - 3276 2021年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemBioChem

Chemical ligation reaction of DNA is useful for the construction of long functional DNA using oligonucleotide fragments that are prepared by solid phase chemical synthesis. However, the unnatural linkage structure formed by the ligation reaction generally impairs the biological function of the resulting ligated DNA. We achieved the complete chemical synthesis of 78 and 258 bp synthetic DNAs via multiple chemical ligation reactions with phosphorothioate and haloacyl-modified DNA fragments. The latter synthetic DNA, coding shRNA for luciferase genes with a designed truncated SV promoter sequence, successfully induced the expected gene silencing effect in HeLa cells.

DOI： 10.1002/cbic.202100312

Antisense Oligonucleotide Modified with Disulfide Units Induces Efficient Exon Skipping in <i>mdx</i> Myotubes through Enhanced Membrane Permeability and Nucleus Internalization 査読有り

Hiraoka, H; Shu, ZM; Le, BT; Masuda, K; Nakamoto, K; Fangjie, L; Abe, N; Hashiya, F; Kimura, Y; Shimizu, Y; Veedu, RN; Abe, H

CHEMBIOCHEM 22 巻 ( 24 ) 頁： 3437 - 3442 2021年10月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemBioChem

We have found that antisense oligonucleotides and siRNA molecules modified with repeat structures of disulfide units can be directly introduced into the cytoplasm and exhibit a suppressive effect on gene expression. In this study, we analyzed the mechanism of cellular uptake of these membrane-permeable oligonucleotides (MPONs). Time-course analysis by confocal microscopy showed that the uptake of MPONs from the plasma membrane to the cytoplasm reached 50 % of the total uptake in about 5 min. In addition, analysis of the plasma membrane proteins to which MPONs bind, identified several proteins, including voltage-dependent anion channel. Next, we analyzed the behavior of MPONs in the cell and found them to be abundant in the nucleus as early as 24 h after addition with the amount increasing further after 48 and 72 h. The amount of MPONs was 2.5-fold higher than that of unmodified oligonucleotides in the nucleus after 72 h. We also designed antisense oligonucleotides and evaluated the effect of MPONs on mRNA exon skipping using DMD model cells; MPONs caused exon skipping with 69 % efficiency after 72 h, which was three times higher than the rate of the control. In summary, the high capacity for intracytoplasmic and nuclear translocation of MPONs is expected to be useful for therapeutic strategies targeting exon skipping.

DOI： 10.1002/cbic.202100413

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1002/cbic.202100413

Variety of Nucleotide Polymerase Mutants Aiming to Synthesize Modified RNA 査読有り国際誌

Ohashi, S; Hashiya, F; Abe, H

CHEMBIOCHEM 22 巻 ( 14 ) 頁： 2398 - 2406 2021年4月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemBioChem

Significant efforts have been made to develop therapeutic RNA aptamers that exploit synthetic RNA to capture target molecules. However, ensuring RNA aptamers are resistant against intrinsic nucleases remains an issue and restricts their use as therapeutics. Introduction of chemical modifications to the 2′ sugar moiety of RNA improves their stability effectively and can be achieved by chemical synthesis using modified phosphoramidites; however, this approach is not suitable for preparing long RNA molecules. Although recombinant nucleotide polymerases can transcribe RNA, these polymerases cannot synthesize modified RNA because they do not recognize 2′ modified nucleoside triphosphates. In this review, we focus on several polymerase mutants that tolerate substrates containing modifications of the 2′ sugar moiety to synthesize RNA, and the problems that must be overcome to prepare chemically modified RNA with high efficacy by in vitro transcription.

DOI： 10.1002/cbic.202100004

Structure, Synthesis and Inhibition Mechanism of Nucleoside Analogues as HIV-1 Reverse Transcriptase Inhibitors (NRTIs) 査読有り国際誌

Yoshida, Y; Honma, M; Kimura, Y; Abe, H

CHEMMEDCHEM 16 巻 ( 5 ) 頁： 743 - 766 2021年3月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemMedChem

Acquired immunodeficiency syndrome (AIDS) is caused by infection with the human immunodeficiency virus (HIV). Although treatments against HIV infection are available, AIDS remains a serious disease that causes many deaths annually. Although a variety of anti-HIV drugs have been synthesized and marketed to treat HIV-infected patients, nucleoside analogue reverse transcriptase inhibitors (NRTIs), which mimic nucleosides, are used extensively and remain a subject of interest to medicinal chemists. However, HIV has acquired drug resistance against NRTIs, and thus the struggle to find novel therapies continues. In this review, we trace the trajectory of NRTIs, focusing on the synthesis, mechanisms of action and applications of NRTIs that have been developed.

DOI： 10.1002/cmdc.202000695

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1002/cmdc.202000695

Chemical Synthesis of Circular RNAs with Phosphoramidate Linkages for Rolling-Circle Translation.

Nakamoto K, Abe H

Current protocols 1 巻 ( 3 ) 頁： e43 2021年3月

化学修飾核酸の合成と核酸医薬への応用

野村浩平, 村瀬裕貴, 中本航介, 木村康明, 阿部奈保子, 安成鎮, 小林芳明, 程久美子, 阿部洋

日本薬学会年会要旨集 141年会巻頁： 27V05 - pm02S 2021年3月

Phosphorothioate Modification of mRNA Accelerates the Rate of Translation Initiation to Provide More Efficient Protein Synthesis 査読有り

Kawaguchi Daisuke, Kodama Ayumi, Abe Naoko, Takebuchi Kei, Hashiya Fumitaka, Tomoike Fumiaki, Nakamoto Kosuke, Kimura Yasuaki, Shimizu Yoshihiro, Abe Hiroshi

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 59 巻 ( 40 ) 頁： 17403 - 17407 2020年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Angewandte Chemie - International Edition

Messenger RNAs (mRNAs) with phosphorothioate modification (PS-mRNA) to the phosphate site of A, G, C, and U with all 16 possible combinations were prepared, and the translation reaction was evaluated using an E. coli cell-free translation system. Protein synthesis from PS-mRNA increased in 12 of 15 patterns when compared with that of unmodified mRNA. The protein yield increased 22-fold when the phosphorothioate modification at A/C sites was introduced into the region from the 5′-end to the initiation codon. Single-turnover analysis of PS-mRNA translation showed that phosphorothioate modification increases the number of translating ribosomes, thus suggesting that the rate of translation initiation (rate of ribosome complex formation) is positively affected by the modification. The method provides a new strategy for improving translation by using non-natural mRNA.

DOI： 10.1002/anie.202007111

Free-Energy Calculation of Ribonucleic Inosines and Its Application to Nearest-Neighbor Parameters

Sakuraba Shun, Iwakiri Junichi, Hamada Michiaki, Kameda Tomoshi, Tsuji Genichiro, Kimura Yasuaki, Abe Hiroshi, Asai Kiyoshi

JOURNAL OF CHEMICAL THEORY AND COMPUTATION 16 巻 ( 9 ) 頁： 5923 - 5935 2020年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Chemical Theory and Computation

Can current simulations quantitatively predict the stability of ribonucleic acids (RNAs)? In this research, we apply a free-energy perturbation simulation of RNAs containing inosine, a modified ribonucleic base, to the derivation of RNA nearest-neighbor parameters. A parameter set derived solely from 30 simulations was used to predict the free-energy difference of the RNA duplex with a mean unbiased error of 0.70 kcal/mol, which is a level of accuracy comparable to that obtained with parameters derived from 25 experiments. We further show that the error can be lowered to 0.60 kcal/mol by combining the simulation-derived free-energy differences with experimentally measured differences. This protocol can be used as a versatile method for deriving nearest-neighbor parameters of RNAs with various modified bases.

DOI： 10.1021/acs.jctc.0c00270

Developments in siRNA Modification and Ligand Conjugated Delivery To Enhance RNA Interference Ability 国際誌

Maguregui Ander, Abe Hiroshi

CHEMBIOCHEM 21 巻 ( 13 ) 頁： 1808 - 1815 2020年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ChemBioChem

There is great potential for siRNA in the treatment of diseases through the reduction of damaging protein translation by RNA interference. However, the delivery and cell uptake of siRNA pose a serious problem in its therapeutic application. Methods to overcome this issue include chemical modification of the siRNA duplex to improve pharmacokinetics, stability and efficacy, and conjugation to small ligand molecules to enable membrane penetration, targetability and potency. In this review, the most common modifications of siRNA will be discussed, along with ligand conjugates that are believed to be the most promising in advancing the field of targeted siRNA delivery.

DOI： 10.1002/cbic.202000009

Chemically synthesized circular RNAs with phosphoramidate linkages enable rolling circle translation 査読有り

Nakamoto Kosuke, Abe Naoko, Tsuji Genichiro, Kimura Yasuaki, Tomoike Fumiaki, Shimizu Yoshihiro, Abe Hiroshi

CHEMICAL COMMUNICATIONS 56 巻 ( 46 ) 頁： 6217 - 6220 2020年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

Circular RNA without a stop codon enables rolling circle translation. To produce circular RNAs, we carried out one-pot chemical synthesis of circular RNA from RNA fragments with the use of an EDC/HOBt-based chemical ligation reaction. The synthesized circular RNAs acted as translation templates, despite the presence of unnatural phosphoramidate linkages.

DOI： 10.1039/d0cc02140g

Quantification of native mRNA dynamics in living neurons using fluorescence correlation spectroscopy and reduction-triggered fluorescent probes 査読有り

Fujita Hirotaka, Oikawa Ryota, Hayakawa Mayu, Tomoike Fumiaki, Kimura Yasuaki, Okuno Hiroyuki, Hatashita Yoshiki, Fiallos Oliveros Carolina, Bito Haruhiko, Ohshima Toshio, Tsuneda Satoshi, Abe Hiroshi, Inoue Takafumi

JOURNAL OF BIOLOGICAL CHEMISTRY 295 巻 ( 23 ) 頁： 7923 - 7940 2020年6月

Disulfide-unit Conjugation Enables Ultrafast Cytosolic Internalization of Antisense DNA and siRNA 査読有り

Nakamoto Kosuke, Shu Zhaoma, Abe Hiroshi

JOURNAL OF SYNTHETIC ORGANIC CHEMISTRY JAPAN 78 巻 ( 5 ) 頁： 456 - 464 2020年5月

Translational control by secondary-structure formation in mRNA in a eukaryotic system 招待有り

Kawaguchi Daisuke, Shimizu Saaya, Abe Naoko, Hashiya Fumitaka, Tomoike Fumiaki, Kimura Yasuaki, Abe Hiroshi

NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS 39 巻 ( 1-3 ) 頁： 195 - 203 2020年2月

SYNTHESIS AND BIOLOGICAL EVALUATION OF NMDI14 DERIVATIVES AS ANTI-MESOTHELIOMA AGENTS 査読有り

Hong Nhung Nguyen, Suzuki Koya, Kimura Yasuaki, Hirokawa Takatsugu, Murakami-Tonami Yuko, Abe Hiroshi

HETEROCYCLES 100 巻 ( 2 ) 頁： 253 - 266 2020年2月

A robust model for quantitative prediction of the silencing efficacy of wild-type and A-to-I edited miRNAs 査読有り

Tian Shen, Terai Goro, Kobayashi Yoshiaki, Kimura Yasuaki, Abe Hiroshi, Asai Kiyoshi, Ui-Tei Kumiko

RNA BIOLOGY 17 巻 ( 2 ) 頁： 264 - 280 2020年2月

Intracellular Delivery of Antisense DNA and siRNA with Amino Groups Masked with Disulfide Units 査読有り

Shu Zhaoma, Ota Azumi, Takayama Yukiya, Katsurada Yuri, Kusamori Kosuke, Abe Naoko, Nakamoto Kosuke, Tomoike Fumiaki, Tada Seiichi, Ito Yoshihiro, Nishikawa Makiya, Kimura Yasuaki, Abe Hiroshi

CHEMICAL & PHARMACEUTICAL BULLETIN 68 巻 ( 2 ) 頁： 129 - 132 2020年2月

Intracellular build-up RNAi with single-strand circular RNAs as siRNA precursors 査読有り

Kimura Yasuaki, Shu Zhaoma, Ito Mika, Abe Naoko, Nakamoto Kosuke, Tomoike Fumiaki, Shuto Satoshi, Ito Yoshihiro, Abe Hiroshi

CHEMICAL COMMUNICATIONS 56 巻 ( 3 ) 頁： 466 - 469 2020年1月

SYNTHESIS AND BIOLOGICAL EVALUATION OF NMDI14 DERIVATIVES AS ANTI-MESOTHELIOMA AGENTS 査読有り

Hong Nhung Nguyen, Suzuki Koya, Kimura Yasuaki, Hirokawa Takatsugu, Murakami-Tonami Yuko, Abe Hiroshi

Heterocycles 100 巻 ( 2 ) 頁： 253 - 266 2020年1月

Intracellular delivery of Antisense DNA and siRNA with amino groups masked with disulfide units 査読有り

Zhaoma Shu, Azumi Ota, Yukiya Takayama, Yuri Katsurada, Kosuke Kusamori, Naoko Abe, Kosuke Nakamoto, Fumiaki Tomoike, Seiichi Tada, Yoshihiro Ito, Makiya Nishikawa, Yasuaki Kimura, Hiroshi Abe

Chemical and Pharmaceutical Bulletins 2020年

Intracellular Build-up RNAi with Single-Strand Circular RNAs as siRNA Precursors 査読有り

Yasuaki Kimura, Zhaoma Shu, Mika Ito, Naoko Abe, Kosuke Nakamoto, Fumiaki Tomoike, Satoshi Shuto, Yoshihiro Ito, Hiroshi Abe

Chemical Communications 2020年

Quantification of native mRNA dynamics in living neurons using fluorescence correlation spectroscopy and reduction-triggered fluorescent probes 査読有り国際誌

Hirotaka Fujita, Ryota Oikawa, Mayu Hayakawa, Fumiaki Tomoike, Yasuaki Kimura, Hiroyuki Okuno, Yoshiki Hatashita, Carolina Fiallos Oliveros, Haruhiko Bito, Toshio Ohshima, Satoshi Tsuneda, Hiroshi Abe, Takafumi Inoue

Journal of Biological Chemistry in press 巻 2020年

Intracellular Delivery of Antisense DNA and siRNA with Amino Groups Masked with Disulfide Units. 査読有り

68 巻 ( 2 ) 頁： 129 - 132 2020年

Synthesis of 1,1,2-trisubstituted cyclopropane nucleosides in enantiomerically pure forms 査読有り国際誌

Fushihara Daichi, Fukuda Hayato, Abe Hiroshi, Shuto Satoshi

NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS 38 巻 ( 12 ) 頁： 921 - 941 2019年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleosides, Nucleotides and Nucleic Acids

Due to the unique rigid and small steric feature of cyclopropane, cyclopropane nucleosides (CPNs) in which the ribose (deoxyribose) of nucleosides are replaced by a hydroxy-substituted cyclopropane, are of great biological interest. Novel 1,1,2-trisubstituted cyclopropane nucleosides were synthesized in enantiomerically pure forms as potential antiviral agents. In the synthesis, two cyclopropane tosylates, which were prepared from chiral cyclopropane lactones previously reported by us, were used effectively as common intermediates for the CPNs. These CPNs are also potentially useful as nucleoside units to incorporate into oligonucleotides in nucleic acids chemotherapy studies.

DOI： 10.1080/15257770.2019.1625380

N-6-methyl adenosine in siRNA evades immune response without reducing RNAi activity 査読有り

Imaeda Akihiro, Tomoike Fumiaki, Hayakawa Mayu, Nakamoto Kosuke, Kimura Yasuaki, Abe Naoko, Abe Hiroshi

38 巻 ( 12 ) 頁： 972 - 979 2019年12月

Translational contro by secondary-structure formation in mRNA in eukaryotic system 査読有り

Daisuke Kawaguchi, Saaya Shimizu, Naoko Abe, Fumitaka Hashiya, Tomoike Fumiaki, Yasuaki Kimura, Hiroshi Abe

Nucleosides, Nucleotides and Nucleic Acids 頁： 1 - 9 2019年9月

RNA imaging by chemical probes 招待有り査読有り

Tomoike Fumiaki, Abe Hiroshi

ADVANCED DRUG DELIVERY REVIEWS 147 巻頁： 44 - 58 2019年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Advanced Drug Delivery Reviews

Sequence-specific detection of intracellular RNA is one of the most important approaches to understand life phenomena. However, it is difficult to detect RNA in living cells because of its variety and scarcity. In the last three decades, several chemical probes have been developed for RNA detection in living cells. These probes are composed of DNA or artificial nucleic acid and hybridize with the target RNA in a sequence-specific manner. This hybridization triggers a change of fluorescence or a chemical reaction. In this review, we classify the probes according to the associated fluorogenic mechanism, that is, interaction between fluorophore and quencher, environmental change of fluorophore, and template reaction with/without ligation. In addition, we introduce examples of RNA imaging in living cells.

DOI： 10.1016/j.addr.2019.08.001

Disulfide-Unit Conjugation Enables Ultrafast Cytosolic Internalization of Antisense DNA and siRNA 査読有り

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 58 巻 ( 20 ) 頁： 6611 - 6615 2019年5月

Disulfide‐Unit Conjugation Enables Ultrafast Cytosolic Internalization of Antisense DNA and siRNA 査読有り

Zhaoma Shu, Iku Tanaka, Azumi Ota, Daichi Fushihara, Naoko Abe, Saki Kawaguchi, Kosuke Nakamoto, Fumiaki Tomoike, Seiichi Tada, Yoshihiro Ito, Yasuaki Kimura, Hiroshi Abe

Angewandte Chemie 131 巻 ( 20 ) 頁： 6683 2019年5月

A Covalent Inhibitor for Glutathione S-Transferase Pi (GSTP(1-1)) in Human Cells 査読有り

Shishido Yuko, Tomoike Fumiaki, Kuwata Keiko, Fujikawa Haruka, Sekido Yoshitaka, Murakami-Tonami Yuko, Kameda Tomoshi, Abe Naoko, Kimura Yasuaki, Shuto Satoshi, Abe Hiroshi

CHEMBIOCHEM 20 巻 ( 7 ) 頁： 900 - 905 2019年4月

膜透過性を有するGST共有結合性阻害剤の開発

友池史明, 宍戸裕子, 藤川遥加, 木村康明, 桑田啓子, 村上優子, 福井健二, 関戸好孝, 矢野貴人, 亀田倫史, 周東智, 阿部洋

日本薬学会年会要旨集 139年会巻 ( 2 ) 頁： 80 - 80 2019年3月

ウイルスタンパク質の活性をトリガーとした新規不可逆阻害剤の開発査読有り

福井竜介, 新美結士, 片倉秀雄, 鈴木哲郎, 村上努, 児玉栄一, 木村康明, 阿部洋

日本薬学会年会要旨集 139年会巻 ( 2 ) 頁： 103 - 103 2019年3月

Disulfide‐unit conjugation enables ultrafast cytosolic internalization of antisense DNA and siRNA 査読有り

Zhaoma Shu, Iku Tanaka, Azumi Ota, Daichi Fushihara, Naoko Abe, Saki Kawaguchi, Kosuke Nakamoto, Fumiaki Tomoike, Seiichi Tada, Yoshihiro Ito, Yasuaki Kimura, Hiroshi Abe

Angewandte Chemie International Edition 58 巻頁： 6611 - 6615 2019年

A Covalent Inhibitor for Glutathione S‐Transferase Pi (GSTP1‐1) in Human Cells 査読有り

Yuko Shishido, Fumiaki Tomoike, Keiko Kuwata, Haruka Fujikawa, Yoshitaka Sekido, Yuko Murakami-Tonami, Tomoshi Kameda, Naoko Abe, Yasuaki Kimura, Satoshi Shuto, Hiroshi Abe

ChemBioChem 20 巻頁： 900 - 905 2019年

N6-methyl adenosine in siRNA evades immune response without reducing RNAi activity 査読有り

Akihiro Imaeda, Fumiaki Tomoike, Mayu Hayakawa, Kosuke Nakamoto, Yasuaki Kimura, Naoko Abe, Hiroshi Abe

Nucleosides, Nucleotides and Nucleic Acids 38 巻頁： 6611 - 6615 2019年

Nearest-neighbor parameter for inosine-cytosine pairs through a combined experimental and computational approach 査読有り

Shun Sakuraba, Junichi Iwakiri, Michiaki Hamada, Tomoshi Kameda, Genichiro Tsuji, Yasuaki Kimura, Hiroshi Abe, Kiyoshi Asai

2018年10月

Structural optimization of pseudorotaxane-forming oligonucleotides for efficient and stable complex formation

Onizuka Kazumitsu, Miyashita Takuya, Chikuni Tomoko, Ozawa Mamiko, Abe Hiroshi, Nagatsugi Fumi

NUCLEIC ACIDS RESEARCH 46 巻 ( 17 ) 頁： 8710 - 8719 2018年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic Acids Research

Interlocked structures, such as rotaxane and catenane, combine both static and dynamic properties. To expand their unique properties into the chemical biology field, a spontaneous formation method of the interlocked structures with the target would be ideal. We have previously developed a pseudorotaxane-forming oligo DNA (prfODN) to spontaneously form topological DNA/RNA architectures. In this study, we report the structural optimization of prfODNs for the efficient and stable complex formation. The optimized prfODNs efficiently formed pseudorotaxane structures with a DNA or RNA target, and the yield for the RNA target reached 85% in 5 min. In addition, the optimized prfODNs could form the pseudorotaxane structure with a smaller ring size and the structure significantly increased the kinetic stability. Furthermore, the catenane structure was successfully formed with the optimized prfODNs to provide the conclusive evidence for the formation of the threaded structure. This information will be valuable for developing new chemical methods using functional nucleic acids for antisense oligo nucleotides and DNA/RNA nanotechnology.

DOI： 10.1093/nar/gky744

MGST1, a GSH transferase/peroxidase essential for development and hematopoietic stem cell differentiation

Brautigam Lars, Zhang Jie, Dreij Kristian, Spahiu Linda, Holmgren Arne, Abe Hiroshi, Tew Kenneth D., Townsend Danyelle M., Kelner Michael J., Morgenstern Ralf, Johansson Katarina

REDOX BIOLOGY 17 巻頁： 171 - 179 2018年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Redox Biology

We show for the first time that, in contrast to other glutathione transferases and peroxidases, deletion of microsomal glutathione transferase 1 (MGST1) in mice is embryonic lethal. To elucidate why, we used zebrafish development as a model system and found that knockdown of MGST1 produced impaired hematopoiesis. We show that MGST1 is expressed early during zebrafish development and plays an important role in hematopoiesis. High expression of MGST1 was detected in regions of active hematopoiesis and co-expressed with markers for hematopoietic stem cells. Further, morpholino-mediated knock-down of MGST1 led to a significant reduction of differentiated hematopoietic cells both from the myeloid and the lymphoid lineages. In fact, hemoglobin was virtually absent in the knock-down fish as revealed by diaminofluorene staining. The impact of MGST1 on hematopoiesis was also shown in hematopoietic stem/progenitor cells (HSPC) isolated from mice, where it was expressed at high levels. Upon promoting HSPC differentiation, lentiviral shRNA MGST1 knockdown significantly reduced differentiated, dedicated cells of the hematopoietic system. Further, MGST1 knockdown resulted in a significant lowering of mitochondrial metabolism and an induction of glycolytic enzymes, energetic states closely coupled to HSPC dynamics. Thus, the non-selenium, glutathione dependent redox regulatory enzyme MGST1 is crucial for embryonic development and for hematopoiesis in vertebrates.

DOI： 10.1016/j.redox.2018.04.013

Chemical ligation reaction for oligonucleotides based on electrophilic phosphorothioester 査読有り

Kimura Yasuaki, Maruyama Hideto, Oikawa Ryota, Hayakawa Mayu, Abe Naoko, Tsuji Genichiro, Matsuda Akira, Shuto Satoshi, Ito Yoshihiro, Abe Hiroshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 255 巻 2018年3月

Development of glutathione S-transferase (GST) covalent inhibitor

Shishido Yuko, Tomoike Fumiaki, Kimura Yasuaki, Kuwata Keiko, Yano Takato, Fukui Kenji, Fujikawa Haruka, Sekido Yoshitaka, Murakami-Tonami Yuko, Kameda Tomoshi, Shuto Satoshi, Abe Hiroshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 255 巻頁： . 2018年3月

Nano structure design for RNA interference

Shu Zhaoma, Abe Naoko, Tomoike Fumiaki, Kimura Yasuaki, Ito Yoshihiro, Abe Hiroshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 255 巻頁： . 2018年3月

Chemical ligation reaction for oligonucleotides based on electrophilic phosphorothioester

Kimura Yasuaki, Maruyama Hideto, Oikawa Ryota, Hayakawa Mayu, Abe Naoko, Tsuji Genichiro, Matsuda Akira, Shuto Satoshi, Ito Yoshihiro, Abe Hiroshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 255 巻頁： . 2018年3月

Chemical Ligation Reactions of Oligonucleotides for Biological and Medicinal Applications

Abe Hiroshi, Kimura Yasuaki

CHEMICAL & PHARMACEUTICAL BULLETIN 66 巻 ( 2 ) 頁： 117-122 2018年2月

Development of GST (Glutathione S-transferase) covalent inhibitor

Shishido Yuko, Sekido Yoshitaka, Murakami-Tonami Yuko, Abe Hiroshi

CANCER SCIENCE 109 巻頁： 374-374 2018年1月

MGST1, a GSH transferase/peroxidase essential for development and hematopoietic stem cell differentiation 査読有り

Lars Bräutigam, Jie Zhang, Kristian Dreij, Linda Spahiu, Arne Holmgren, Hiroshi Abe, Kenneth D. Tew, Danyelle M. Townsend, Michael J. Kelner, Ralf Morgenstern, Katarina Johansson

Redox Biology 17 巻頁： 171 - 179 2018年

Preparation of Circular RNA In Vitro 査読有り

Abe Naoko, Kodama Ayumi, Abe Hiroshi

CIRCULAR RNAS: METHODS AND PROTOCOLS 1724 巻頁： 181 - 192 2018年

記述言語：英語掲載種別：論文集(書籍)内論文出版者・発行元：Methods in Molecular Biology

This chapter describes a simple and straightforward way to obtain single-stranded circular RNA sequences in vitro. Linear RNA that is phosphorylated at the 5′ end is first prepared by a chemical or enzymatic method, then circularized using ligase. The function of the prepared circular RNA molecule, such as an ability to induce translation, can then be investigated.

DOI： 10.1007/978-1-4939-7562-4_15

Chemical Ligation Reactions of Oligonucleotides for Biological and Medicinal Applications.

Abe H, Kimura Y

Chemical & pharmaceutical bulletin 66 巻 ( 2 ) 頁： 117 - 122 2018年

Structural optimization of pseudorotaxane-forming oligonucleotides for efficient and stable complex formation 査読有り

Kazumitsu Onizuka, Takuya Miyashita, Tomoko Chikuni, Mamiko Ozawa, Hiroshi Abe, Fumi Nagatsugi

Nucleic Acids Research 46 巻頁： 8710 - 8719 2018年

グルタチオン-S-トランスフェラーゼを標的とした共有結合性阻害剤の開発

宍戸裕子, 藤川遥加, 友池史明, 木村康明, 桑田啓子, 村上優子, 福井健二, 関戸好孝, 矢野貴人, 周東智, 阿部洋

生命科学系学会合同年次大会 2017年度巻頁： [3P - 0205] 2017年12月

A covalent G-site inhibitor for glutathione S-transferase Pi (GSTP(1-1)) 査読有り

Shishido Yuko, Tomoike Fumiaki, Kimura Yasuaki, Kuwata Keiko, Yano Takato, Fukui Kenji, Fujikawa Haruka, Sekido Yoshitaka, Murakami-Tonami Yuko, Kameda Tomoshi, Shuto Satoshi, Abe Hiroshi

CHEMICAL COMMUNICATIONS 53 巻 ( 81 ) 頁： 11138 - 11141 2017年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

We herein report the first covalent G-site-binding inhibitor for GST, GS-ESF (1), which irreversibly inhibited the GSTP1-1 function. LC-MS/MS and X-ray structure analyses of the covalently linked GST-inhibitor complex suggested that 1 reacted with Tyr108 of GSTP1-1. The mechanism of covalent bond formation was discussed based on MD simulation results.

DOI： 10.1039/c7cc05829b

Tethered 1,2-Si-Group Migrations in Radical-Mediated Ring Enlargements of Cyclic Alkoxysilanes: An EPR Spectroscopic and Computational Investigation

Walton John C., Kanada Ryutaro, Iwamoto Takeaki, Shuto Satoshi, Abe Hiroshi

JOURNAL OF ORGANIC CHEMISTRY 82 巻 ( 13 ) 頁： 6886-6894 2017年7月

Chemical ligation of oligonucleotides using an electrophilic phosphorothioester 査読有り

Hideto Maruyama, Ryota Oikawa, Mayu Hayakawa, Shono Takamori, Yasuaki Kimura, Naoko Abe, Genichiro Tsuji, Akira Matsuda, Satoshi Shuto, Yoshihiro Ito, Hiroshi Abe

Nucleic Acids Research 45 巻 ( 12 ) 頁： 7042-7048 2017年7月

Enhancement of synergistic gene silencing by RNA interference using branched "3-in-1" trimer siRNA

Nair Baiju G., Zhou Yue, Hagiwara Kyoji, Ueki Masashi, Isoshima Takashi, Abe Hiroshi, Ito Yoshihiro

JOURNAL OF MATERIALS CHEMISTRY B 5 巻 ( 22 ) 頁： 4044-4051 2017年6月

Pseudorotaxane formation via the slippage process with chemically cyclized oligonucleotides

Onizuka Kazumitsu, Chikuni Tomoko, Amemiya Takuya, Miyashita Takuya, Onizuka Kyoko, Abe Hiroshi, Nagatsugi Fumi

NUCLEIC ACIDS RESEARCH 45 巻 ( 9 ) 頁： 5036-5047 2017年5月

Enhancement of synergistic gene silencing by RNA interference using branched “3-in-1” trimer siRNA 査読有り

5 巻頁： 4044-4051 2017年5月

環状RNAを用いたローリングサークル型翻訳反応

阿部奈保子, 阿部洋

生物物理 57 巻 ( 1 ) 頁： 5 - 10 2017年

記述言語：日本語出版者・発行元：一般社団法人日本生物物理学会

<p>In an <i>E. coli</i> cell-free translation system, we found that a circular RNA containing an infinite open reading frame produced more translation product than its linear counterpart by two orders of magnitude, because a ribosome can work more effectively towards the elongation on circular RNA than it can on linear RNA. We then tested circular RNAs containing an infinite open reading frame could be translated in eukaryotic systems, in the absence of any particular element for internal ribosome entry, a poly-A tail, or a cap structure. We found that the circular RNAs also produced long peptides in eukaryotic translation systems, possibly owing to the rolling circle amplification mechanism.</p>

DOI： 10.2142/biophys.57.005

Design and Synthesis of Cyclopropane Congeners of Resolvin E2, an Endogenous Proresolving Lipid Mediator, as Its Stable Equivalents 査読有り

Hayato Fukuda, Ryuta Muromoto, Yuuki Takakura, Kohei Ishimura, Ryutaro Kanada, Daichi Fushihara, Makoto Tanabe, Kotaro Matsubara, Toru Hirao, Koki Hirashima, Hiroshi Abe, Mitsuhiro Arisawa, Tadashi Matsuda, Satoshi Shuto

Organic Letters 18 巻 ( 24 ) 頁： 6224 - 6227 2016年12月

掲載種別：研究論文（学術雑誌）

© 2016 American Chemical Society. Lipid chemical mediator resolvins with highly potent anti-inflammatory activity can be leads to develop novel anti-inflammatory drugs; however, they are unstable in oxygen due to their characteristic polyunsaturated structures. To solve the problem, CP-RvE2 has been designed and synthesized in which the cis-olefin of RvE2 was replaced with a cyclopropane. CP-RvE2s were much more stable than RvE2 against autoxidation and equipotent or more potent than RvE2. CP-RvE2s were successfully identified as stable equivalents of RvE2.

DOI： 10.1021/acs.orglett.6b02612

Catalytic Conversion of Lipophilic Substrates by Phase constrained Enzymes in the Aqueous or in the Membrane Phase 査読有り

Marcus Cebula. Ilke Simek Turan, Birgitta Sjodin, Madhuranayaki Thulasingam, Joseph Brock, Volodymyr Chmyrov, Jerker Widengren, Hiroshi Abe, Bengt Mannervik, Jesper Haeggstrom, Agnes Rinaldo-Matthis, Engin Akkaya, Ralf Morgenstern

Scientific Reports 6 巻頁： 38316 2016年12月

Design and Synthesis of Cyclopropane Congeners of Resolvin E2, an Endogenous Proresolving Lipid Mediator, as Its Stable Equivalents 査読有り

Hayato Fukuda, Ryuta Muromoto, Yuuki Takakura, Kohei Ishimura, Ryutaro Kanada, Daichi Fushihara, Makoto Tanabe, Kotaro Matsubara, Toru Hirao, Koki Hirashima, Hiroshi Abe, Mitsuhiro Arisawa, Tadashi Matsuda, Satoshi Shuto

Organic Letters 18 巻頁： 6224-6227 2016年11月

Chemical Reactivity Window Determines Prodrug Efficiency toward Glutathione Transferase Overexpressing Cancer Cells 査読有り

van Gisbergen Marike W., Cebula Marcus, Zhang Jie, Ottosson-Wadlund Astrid, Dubois Ludwig, Lambin Philippe, Tew Kenneth D., Townsend Danyelle M., Haenen Guido R. M. M., Drittij-Repders Marie-Jose, Saneyoshi Hisao, Araki Mika, Shishido Yuko, Ito Yoshihiro, Arner Elias S. J., Abe Hiroshi, Morgenstern Ralf, Johansson Katarina

MOLECULAR PHARMACEUTICS 13 巻 ( 6 ) 頁： 2010 - 2025 2016年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Molecular Pharmaceutics

Glutathione transferases (GSTs) are often overexpressed in tumors and frequently correlated to bad prognosis and resistance against a number of different anticancer drugs. To selectively target these cells and to overcome this resistance we previously have developed prodrugs that are derivatives of existing anticancer drugs (e.g., doxorubicin) incorporating a sulfonamide moiety. When cleaved by GSTs, the prodrug releases the cytostatic moiety predominantly in GST overexpressing cells, thus sparing normal cells with moderate enzyme levels. By modifying the sulfonamide it is possible to control the rate of drug release and specifically target different GSTs. Here we show that the newly synthesized compounds, 4-acetyl-2-nitro-benzenesulfonyl etoposide (ANS-etoposide) and 4-acetyl-2-nitro-benzenesulfonyl doxorubicin (ANS-DOX), function as prodrugs for GSTA1 and MGST1 overexpressing cell lines. ANS-DOX, in particular, showed a desirable cytotoxic profile by inducing toxicity and DNA damage in a GST-dependent manner compared to control cells. Its moderate conversion of 500 nmol/min/mg, as catalyzed by GSTA1, seems hereby essential since the more reactive 2,4-dinitrobenzenesulfonyl doxorubicin (DNS-DOX) (14000 nmol/min/mg) did not display a preference for GSTA1 overexpressing cells. DNS-DOX, however, effectively killed GSTP1 (20 nmol/min/mg) and MGST1 (450 nmol/min/mg) overexpressing cells as did the less reactive 4-mononitrobenzenesulfonyl doxorubicin (MNS-DOX) in a MGST1-dependent manner (1.5 nmol/min/mg) as shown previously. Furthermore, we show that the mechanism of these prodrugs involves a reduction in GSH levels as well as inhibition of the redox regulatory enzyme thioredoxin reductase 1 (TrxR1) by virtue of their electrophilic sulfonamide moiety. TrxR1 is upregulated in many tumors and associated with resistance to chemotherapy and poor patient prognosis. Additionally, the prodrugs potentially acted as a general shuttle system for DOX, by overcoming resistance mechanisms in cells. Here we propose that GST-dependent prodrugs require a conversion rate "window" in order to selectively target GST overexpressing cells, while limiting their effects on normal cells. Prodrugs are furthermore a suitable system to specifically target GSTs and to overcome various drug resistance mechanisms that apply to the parental drug.

DOI： 10.1021/acs.molpharmaceut.6b00140

Chemical reactivity window determines prodrug efficiency towards glutathione transferase overexpressing cancer cells 査読有り

Marie van Gisbergen, Marcus Cebula, Jie Zhang, Astrid Ottosson-Wadlund, Ludwig Dubois, Philippe Lambin, Kenneth D. Tew, Danyelle Townsend, Guido Haenen, Marie-Jose Drittij-Reijnders, Hisao Saneyoshi, Mika Araki, Yuko Shishido, Yoshihiro Ito, Elias Arner, Hiroshi Abe, Ralf Morgenstern, Katarina Johansson

Molecular Pharmaceutics 13 巻頁： 2010-2025 2016年4月

Diastereoselective Synthesis of 6 ''-(Z)- and 6 ''-(E)-Fluoro Analogues of Anti-hepatitis B Virus Agent Entecavir and Its Evaluation of the Activity and Toxicity Profile of the Diastereomers 査読有り国際誌

Kumamoto Hiroki, Fukano Misato, Nakano Tomohiko, Iwagami Keito, Takeyama Chiaki, Kohgo Satoru, Imoto Shuhei, Amano Masayuki, Kuwata-Higashi Nobuyo, Aoki Manabu, Abe Hiroshi, Mitsuya Hiroaki, Fukuhara Kiyoshi, Haraguchi Kazuhiro

JOURNAL OF ORGANIC CHEMISTRY 81 巻 ( 7 ) 頁： 2827 - 2836 2016年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Organic Chemistry

A method for the diastereoselective synthesis of 6″-(Z)- and 6″-(E)-fluorinated analogues of the anti-HBV agent entecavir has been developed. Construction of the methylenecyclopentane skeleton of the target molecules has been accomplished by radical-mediated 5-exo-dig cyclization of the selenides 6 and 15 having the phenylsulfanylethynyl structure as a radical accepting moiety. In the radical reaction of the TBS-protected precursor 6, (Z)-anti-12 was formed as a major product. On the other hand, TIPS-protected 15 gave (E)-anti-12. The sulfur-extrusive stannylation of anti-12 furnished a mixture of geometric isomers of the respective vinylstannane, whereas benzoyl-protected 17 underwent the stannylation in the manner of retention of configuration. Following XeF2-mediated fluorination, introduction of the purine base and deoxygenation of the resulting carbocyclic guanosine gave the target (E)- and (Z)-3 after deprotection. Evaluation of the anti-HBV activity of 3 revealed that fluorine-substitution at the 6″-position of entecavir gave rise to a reduction in the cytotoxicity in HepG2 cells with retention of the antiviral activity.

DOI： 10.1021/acs.joc.6b00105

Effect of sigma-pi conjugation between Si-Si bond and pyridine ring in tris(trimethylsilyl) silylpyridine on its spectroscopic property and SN2 reaction with methyl iodide 査読有り

Ichinose Wataru, Abe Hiroshi, Shuto Satoshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 251 巻 2016年3月

Effect of sigma-pi conjugation between Si-Si bond and pyridine ring in tris(trimethylsilyl) silylpyridine on its spectroscopic property and SN2 reaction with methyl iodide

Ichinose Wataru, Abe Hiroshi, Shuto Satoshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 251 巻 2016年3月

Rolling Circle Translation of Circular RNA in Living Human Cells 査読有り

Abe Naoko, Matsumoto Ken, Nishihara Mizuki, Nakano Yukiko, Shibata Aya, Maruyama Hideto, Shuto Satoshi, Matsuda Akira, Yoshida Minoru, Ito Yoshihiro, Abe Hiroshi

SCIENTIFIC REPORTS 5 巻頁： 16435 2015年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Scientific Reports

We recently reported that circular RNA is efficiently translated by a rolling circle amplification (RCA) mechanism in a cell-free Escherichia coli translation system. Recent studies have shown that circular RNAs composed of exonic sequences are abundant in human cells. However, whether these circular RNAs can be translated into proteins within cells remains unclear. In this study, we prepared circular RNAs with an infinite open reading frame and tested their translation in eukaryotic systems. Circular RNAs were translated into long proteins in rabbit reticulocyte lysate in the absence of any particular element for internal ribosome entry, a poly-A tail, or a cap structure. The translation systems in eukaryote can accept much simpler RNA as a template for protein synthesis by cyclisation. Here, we demonstrated that the circular RNA is efficiently translated in living human cells to produce abundant protein product by RCA mechanism. These findings suggest that translation of exonic circular RNAs present in human cells is more probable than previously thought.

DOI： 10.1038/srep16435

Nickel-catalyzed Suzuki-Miyaura coupling of a tertiary iodocyclopropane with wide boronic acid substrate scope: Coupling reaction outcome depends on radical species stability 査読有り

Keisuke Yotsuji, Naoyuki Hoshiya, Takaaki Kobayashi, Hayato Fukuda, Hiroshi Abe, Mitsuhiro Arisawa, Satoshi Shuto

Advanced Synthesis and Catalysis 357 巻 ( 5 ) 頁： 1022 - 1028 2015年3月

掲載種別：研究論文（学術雑誌）

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. We describe a nickel-catalyzed Suzuki-Miyaura arylation of a tertiary iodocyclopropane with arylboronic acids; this is an efficient and convergent strategy for providing various enantioenriched arylcyclopropanes with a quaternary stereogenic center. This is the first metal-catalyzed coupling between a tertiary alkyl electrophile and a wide range of aromatics, including heteroaromatics. We found that the outcome of the Ni-catalyzed coupling with halides as electrophiles was dependent on the stability of the radical species formed during the reaction. The use of tert-butyl alcohol (t-BuOH) as the reaction solvent was very effective, because of its stability under the radical-generating reaction conditions.

DOI： 10.1002/adsc.201401000

Nanostructured RNAs for RNA Interference 査読有り

Nakashima Yuko, Abe Naoko, Ito Yoshihiro, Abe Hiroshi

RNA INTERFERENCE: CHALLENGES AND THERAPEUTIC OPPORTUNITIES 1218 巻頁： 17 - 36 2015年

記述言語：英語掲載種別：論文集(書籍)内論文出版者・発行元：RNA Interference: Challenges and Therapeutic Opportunities

We synthesized three types of nanostructured RNAs that induce RNA interference (RNAi): branched RNAs, dumbbell-shaped RNA, and circular double-stranded RNAs. All three nanostructured RNAs were transformed into double-stranded RNA of approximately 20 base pairs when they were treated with nuclease enzymes such as Dicer. These dsRNA species induced gene silencing when they are were introduced into mammalian cells.

DOI： 10.1007/978-1-4939-1538-5_2

Phosphorylated 5-ethynyl-2 '-deoxyuridine for advanced DNA labeling 査読有り

Seo Siyoong, Onizuka Kazumitsu, Nishioka Chieko, Takahashi Eiki, Tsuneda Satoshi, Abe Hiroshi, Ito Yoshihiro

ORGANIC & BIOMOLECULAR CHEMISTRY 13 巻 ( 15 ) 頁： 4589 - 4595 2015年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Organic and Biomolecular Chemistry

The representative DNA-labeling agent 5-ethynyl-2′-deoxyuridine (EdU) was chemically modified to improve its function. Chemical monophosphorylation was expected to enhance the efficiency of the substrate in DNA polymerization by circumventing the enzymatic monophosphorylation step that consumes energy. In addition, to enhance cell permeability, the phosphates were protected with bis-pivaloyloxymethyl that is stable in buffer and plasma, and degradable inside various cell types. The phosphorylated EdU (PEdU) was less toxic than EdU, and had the same or a slightly higher DNA-labeling ability in vitro. PEdU was also successfully applied to DNA labeling in vivo. In conclusion, PEdU can be used as a less toxic DNA-labeling agent for studies that require long-term cell survival or very sensitive cell lines.

DOI： 10.1039/c5ob00199d

Synthesis of 3’,4’-difluoro-3’-deoxyribonucleosides and its evaluation of the biological activities: Discovery of a novel type of anti-HCV agent 3’,4’-difluorocordycepin 査読有り

Hisashi Shimada, Kazuhiro Haraguchi, Kumi Hotta, Tomoko Miyaike, Yasuyuki Kitagawa, Hiromichi Tanaka, Ryutaro Kaneda, Hiroshi Abe, Satoshi Shuto, Kyoko Mori, Youki Ueda, Nobuyuki Kato, Robert Snoeck, Graciela Andrei, an Balzarini

Bioorganic & Medicinal Chemistry 22 巻 ( 21 ) 頁： 6174 - 6182 2014年11月

Fluorogenic probes using 4-substituted-2-nitrobenzenesulfonyl derivatives as caging groups for the analysis of human glutathione transferase catalyzed reactions (vol 138, pg 7326, 2013) 査読有り

Aya Shibata, Yukiko Nakano, Mika Ito, Mika Araki, Jie Zhang, Yasuhiko Yoshida, Satoshi Shuto, Bengt Mannervik, Ralf Morgenstern, Yoshihiro Ito, Hiroshi Abe

ANALYST 139 巻 ( 17 ) 頁： 4382 - 4382 2014年9月

Conformationally Restricted GABA with Bicyclo[3.1.0]hexane Backbone as the First Highly Selective BGT‑1 Inhibitor 査読有り

Takaaki Kobayashi, Akihiro Suemasa, Arisa Igawa, Soichiro Ide, Hayato Fukuda, Hiroshi Abe, Mitsuhiro Arisawa, Masabumi Minami, Satoshi Shuto

ACS Medicinal Chemistry Letters 5 巻 ( 8 ) 頁： 889 - 893 2014年8月

Sustained delivery of siRNA from dopamine-coated stainless steel surfaces (vol 9, pg 6753, 2013)

Joddar Binata, Albayrak Aydin, Kang Jeonghwa, Nishihara Mizuki, Abe Hiroshi, Ito Yoshihiro

ACTA BIOMATERIALIA 10 巻 ( 8 ) 頁： 3811 - 3811 2014年8月

Development of Molecular Probe Targeting on Glutathion Transferase 査読有り

Ito Mika, Shuto Satoshi, Ito Yoshihiro, Abe Hiroshi

JOURNAL OF SYNTHETIC ORGANIC CHEMISTRY JAPAN 72 巻 ( 7 ) 頁： 822 - 831 2014年7月

記述言語：日本語出版者・発行元：Yuki Gosei Kagaku Kyokaishi/Journal of Synthetic Organic Chemistry

Glutathione S-transferase (GST) are phase II enzymes that catalyze the nucleophilic attack of glutathione (GSH) to a wide range of hydrophobic and electrophilic compounds. GST is of high clinical interest in inflammation, pathophysiology and tumor drug resistance. In addition, GST is known to highly express in cancer cells. Therefore, we tried to develop new luminescent probes that can detect and image GST expression in living cells. Recently, we found that the electrophilic centre in the dinitrobenzenesulfonamide derivative is attacked by GSH and that this reaction is catalyzed by GST releasing the strong fluorophore. Based on this knowledge, we synthesized fluorescent probe, 19F MKI probe, and bioluminogenic probe. The fluorescent probe and kinetic parameters for purified GST were giving a rate enhancement of 106 compared with the nonenzymatic reaction. In addition, fluorescent probe successfully image GST in living cells. The fluorescent probe is potentially useful reagent for detection of GST activity in living cells. 19F MKI and bioluminogenic probes were synthesized using 4-acetyl-2-nitrobenzenesulfonyl group. 19F MKI probe exhibited a peak shift of 19F MRI signal and fluorescence enhancement in a bimodal way in response to GST activity. Bioluminogenic probe provided strong bioluminescence from GST activity. Moreover, both probes were successfully applied to the monitoring of GST expression in living E. coli cells. Successful detection of GST activity in living cells could be very useful for identifying tumor cells that overexpress GSTs.

DOI： 10.5059/yukigoseikyokaishi.72.822

Automatic Pseudorotaxane Formation Targeting on Nucleic Acids Using a Pair of Reactive Oligodeoxynucleotides 査読有り

Onizuka Kazumitsu, Nagatsugi Fumi, Ito Yoshihiro, Abe Hiroshi

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 136 巻 ( 20 ) 頁： 7201 - 7204 2014年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of the American Chemical Society

Here we report a novel method to form a pseudorotaxane architecture using only a pair of reactive oligodeoxyribonucleotides (ODNs), which we designed and synthesized, and then performed the pseudorotaxane formation reaction with both DNA and RNA oligonucleotides. The reaction proceeded smoothly without any extra reagents at 37°C and pH 7.2, leading to the formation of a stable complex on a denaturing polyacrylamide gel. Interestingly, the pseudorotaxane was formed with the cyclized ODN reversibly by the slipping process. This new pseudorotaxane formation represents a promising method for developing new DNA nanotechnologies and antisense oligonucleotides. © 2014 American Chemical Society.

DOI： 10.1021/ja5018283

Development of rapid DNA templated reaction based on nucleophilic aromatic substitution reaction

Shibata Aya, Abe Hiroshi, Uzawa Takanori, Nakano Yukiko, Shuto Satoshi, Ito Yoshihiro

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 247 巻 2014年3月

Synthesis of active siRNA species via chemical ligation

Maruyama Hideto, Matsuda Akira, Shuto Satoshi, Ito Yoshihiro, Abe Hiroshi

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 247 巻 2014年3月

Development of rapid DNA templated reaction based on nucleophilic aromatic substitution reaction 査読有り

Aya Shibata, Hiroshi Abe, Takanori Uzawa, Yukiko Nakano, Satoshi Shuto, Yoshihiro Ito

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 247 巻 2014年3月

Synthesis of active siRNA species via chemical ligation 査読有り

Hideto Maruyama, Akira Matsuda, Satoshi Shuto, Yoshihiro Ito, Hiroshi Abe

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 247 巻 2014年3月

Triphenylphosphinecarboxamide: an effective reagent for the reduction of azides and its application to nucleic acid detection. 査読有り

Saneyoshi H, Ochikubo T, Mashimo T, Hatano K, Ito Y, Abe H

Organic letters 16 巻 ( 1 ) 頁： 30-3 2014年1月

An intracellular buildup reaction of active siRNA species from short RNA fragments 査読有り

Maruyama Hideto, Nakashima Yuko, Shuto Satoshi, Matsuda Akira, Ito Yoshihiro, Abe Hiroshi

CHEMICAL COMMUNICATIONS 50 巻 ( 11 ) 頁： 1284 - 1287 2014年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

Here we report a new strategy for the buildup reaction of active siRNA species from short RNA fragments in living cells using a chemical ligation reaction. This strategy could decrease undesired immune responses and provide more latitude for RNAi technology in the design and concentration of introduced RNA compared to traditional siRNA methods. © 2014 The Royal Society of Chemistry.

DOI： 10.1039/c3cc47529h

Transmembrane molecular transport through nanopores formed by protein nanotubes 査読有り

Nair Baiju G., Nakano Yukiko, Ito Yoshihiro, Abe Hiroshi

CHEMICAL COMMUNICATIONS 50 巻 ( 5 ) 頁： 602 - 604 2014年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

Protein nanotubes formed by layer-by-layer (LbL) assembly can penetrate cells and act as nanopores for direct transmembrane delivery of chemical compounds. © 2013 The Royal Society of Chemistry.

DOI： 10.1039/c3cc45907a

非酵素的に高効率シグナル増幅を可能とする新規RNA検出法を開発

柴田綾、鵜澤尊規、伊藤嘉浩、周東智、阿部洋

化学と生物 52 巻 ( 11 ) 頁： 771-776 2014年

非酵素的に高効率シグナル増幅を可能とする新規RNA検出法を開発

柴田綾, 鵜澤尊規, 伊藤嘉浩, 周東智, 阿部洋

化学と生物 52 巻 ( 11 ) 頁： 771 - 776 2014年

Nanostructured RNAs for RNA interference

Yuko Nakashima, Naoko Abe, Yoshihiro Ito, Hiroshi Abe

Humana Press New York 頁：全 2014年

グルタチオントランスフェラーゼを標的とした分子プローブの開発

伊藤美香,周東智,伊藤嘉浩,阿部洋

有機合成化学教会誌頁： 822 2014年

Polycation-assisted DNA detection by reduction triggered fluorescence amplification probe 査読有り

Saneyoshi Hisao, Shimada Naohiko, Maruyama Atsushi, Ito Yoshihiro, Abe Hiroshi

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 23 巻 ( 24 ) 頁： 6851 - 6853 2013年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

We have developed a fluorescence detection system for DNA, assisted by a comb-type cationic polymer (PLL-g-DX), for accelerating the reaction turnover. The combination of fluorogenic DNA probes with a comb-type cationic polymer has been demonstrated to be an effective means of signal amplification during the detection process. The method described herein represents a simple and enzyme-free detection. © 2013 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmcl.2013.10.005

Synthesis of novel 4′-C-methyl-1′,3′-dioxolane pyrimidine nucleosides and evaluation of its anti-HIV-1 activity 査読有り

Yutaka Kubota, Yuri Kaneda, Kazuhiro Haraguchi, Mirei Mizuno, Hiroshi Abe, Satoshi Shuto, Takayuki Hamasaki, Masanori Baba, Hiromichi Tanaka

Tetrahedron 69 巻 ( 51 ) 頁： 10884 - 10892 2013年12月

An epidermal growth factor derivative with binding affinity for hydroxyapatite and titanium surfaces 査読有り

Kang Jeonghwa, Tada Seiichi, Sakuragi Makoto, Abe Hiroshi, Ito Reiko, Ishikawa Junko, Kurata Shino, Kitajima Takashi, Son Tae Il, Aigaki Toshiro, Ito Yoshihiro

BIOMATERIALS 34 巻 ( 38 ) 頁： 9747 - 9753 2013年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Biomaterials

An epidermal growth factor (EGF) derivative with affinity for apatite and titanium surfaces was designed using a peptide moiety derived from salivary statherin, a protein that adheres to hydroxyapatite. Since the active sequence has two phosphoserine residues, the EGF derivative was prepared by organic synthesis, and a 54 residue peptide was successfully prepared using this method. Circular dichroism spectra indicated that the conformation of EGF was not significantly altered by the addition of the affinity peptide sequence and the mitogenic activity was only slightly reduced when compared with the wild-type protein. However, the binding affinity of the modified EGF to hydroxyapatite and titanium was significantly higher than the unmodified EGF. The phosphate groups in the affinity sequence contributed to the affinity of modified EGF to both apatite and titanium. The modified EGF significantly enhanced the growth of cells on hydroxyapatite and titanium. It was also demonstrated that the bound EGF enhanced the signal transduction for longer periods than unbound EGF. In conclusion, the modified EGF had significantly higher binding affinity for apatite and titanium than soluble EGF, and the bound EGF significantly enhanced cell growth by long-lasting activation of intracellular signal transduction. © 2013 Elsevier Ltd.

DOI： 10.1016/j.biomaterials.2013.09.004

PEGylated antibodies and DNA in organic media and genetic PEGylation

Tada S., Abe H., Ito Y.

ACS Symposium Series 1144 巻頁： 223 - 233 2013年11月

出版者・発行元：ACS Symposium Series

Polyethylene glycol (PEG) is an amphiphilic polymer that is soluble in both water and organic media and can add many functions to biopolymers by conjugation. Here, PEG was coupled to antibodies or oligonucleotides for solubilization into organic media. The former interacted with antigens and the latter formed specific structures and had catalytic activity in organic media. In addition, we attempted to incorporate PEG genetically into a polypeptide. Although the PEG incorporation ratio decreased as the molecular weight of PEG was increased, PEG with a molecular weight of ≤1000 Da was incorporated successfully. The potential applications of PEGylated biopolymers and the methodology for gene PEGylation are discussed. © 2013 American Chemical Society.

DOI： 10.1021/bk-2013-1144.ch016

Very Rapid DNA-Templated Reaction for Efficient Signal Amplification and Its Steady-State Kinetic Analysis of the Turnover Cycle 査読有り

Shibata Aya, Uzawa Takanori, Nakashima Yuko, Ito Mika, Nakano Yukiko, Shuto Satoshi, Ito Yoshihiro, Abe Hiroshi

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 135 巻 ( 38 ) 頁： 14172 - 14178 2013年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of the American Chemical Society

Oligonucleotide-templated reactions are powerful tools for the detection of nucleic acid sequences. One of the major scientific challenges associated with this technique is the rational design of non-enzyme-mediated catalytic templated reactions capable of multiple turnovers that provide high levels of signal amplification. Herein, we report the development of a nucleophilic aromatic substitution reaction-triggered fluorescent probe. The probe underwent a rapid templated reaction without any of the undesired background reactions. The fluorogenic reaction conducted in the presence of a template provided a 223-fold increase in fluorescence after 30 s compared with the nontemplated reaction. The probe provided an efficient level of signal amplification that ultimately enabled particularly sensitive levels of detection. Assuming a simple model for the templated reactions, it was possible to estimate the rate constants of the chemical reaction in the presence and in the absence of the template. From these kinetic analyses, it was possible to confirm that an efficient turnover cycle had been achieved, on the basis of the dramatic enhancement in the rate of the chemical reaction considered to be the rate-determining step. With maximized turnover efficiency, it was demonstrated that the probe could offer a high turnover number of 1500 times to enable sensitive levels of detection with a detection limit of 0.5 pM in the catalytic templated reactions. © 2013 American Chemical Society.

DOI： 10.1021/ja404743m

Long-Lived Luminogenic Probe for Detection of RNA in a Crude Solution of Living Bacterial Cells 査読有り

Saneyoshi Hisao, Ito Yoshihiro, Abe Hiroshi

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 135 巻 ( 37 ) 頁： 13632 - 13635 2013年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of the American Chemical Society

A pre-type sensitizer for a lanthanide complex on an oligonucleotide was successfully converted to a perfect final structure in a target DNA/RNA-templated reaction, without any chemical reagent or enzyme, under neutral conditions. The final form of the lanthanide-oligonucleotide provided a long-lived luminescence signal, appropriate for time-gated luminescence analysis and signal amplification. Target DNA/RNA-assisted time-gated luminescence analysis is a powerful tool for elimination of autofluorescence and detection of target RNA in living bacterial cells. © 2013 American Chemical Society.

DOI： 10.1021/ja406724k

Sustained delivery of siRNA from dopamine-coated stainless steel surfaces 査読有り

Joddar Binata, Albayrak Aydin, Kang Jeonghwa, Nishihara Mizuki, Abe Hiroshi, Ito Yoshihiro

ACTA BIOMATERIALIA 9 巻 ( 5 ) 頁： 6753 - 6761 2013年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Acta Biomaterialia

Dopamine, an adhesive protein can be covalently deposited onto biomaterials. In this study, we evaluated the ability of dopamine-coated surfaces for small interfering RNA (siRNA) immobilization and release. Dopamine was deposited onto 316L stainless steel discs either as a monolayer at acidic pH or as polydopamine at alkaline pH, following which siRNA was immobilized onto these discs. To investigate the RNA interference ability of immobilized siRNA, reduction of luciferase expression in HeLa, and reduction of Egr-1 expression and cell proliferation in human aortic smooth muscle cells (HAoSMCs) were determined. Dopamine treatment of 316L stainless steel discs under both the acidic and alkaline conditions resulted in the deposition of amino (NH 2) groups, which enabled electrostatic immobilization of siRNA. The immobilized siRNA was released from both types of coatings, and enhanced the percent suppression of firefly luciferase activity of HeLa significantly up to ∼96.5% compared to HeLa on non-dopamine controls (18%). Both the release of siRNA and the percent suppression of firefly luciferase activity were sustained for at least 7 days. In another set of experiments, siRNA sequences targeting to inhibit the activity of the transcription factor Egr-1 were eluted from dopamine-coated surfaces to HAoSMCs. Egr-1 siRNA eluted from dopamine-coated surfaces, significantly reduced the proliferation of HAoSMCs and their protein expression of Egr-1. Therefore, this method of surface immobilization of siRNA onto dopamine-coated surfaces might be effective for nucleic acid delivery from stents. © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.actbio.2013.01.007

Nano-sized Molecule: New Approach for Diagnosis and Drug Discovery Foreword 査読有り

Abe Hiroshi, Mizukami Shin

YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 133 巻 ( 3 ) 頁： 349 - 349 2013年3月

Nanostructured RNA for RNA Intereference 査読有り

Abe Hiroshi

YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 133 巻 ( 3 ) 頁： 373 - 378 2013年3月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：Yakugaku Zasshi

RNA interference (RNAi) is a potent and highly specific gene-silencing phenomenon which is initiated or triggered by double-stranded RNAs (dsRNAs). Shortly after the development of RNAi, small interfering RNAs (siRNAs) that are 21 nucleotides in length with a 3′ nucleotide overhang were shown to be very effective in mammalian cells. Much effort has been dedicated to the application of siRNAs, both as biological tools and as therapeutic agents. Currently, synthetic siRNA would be the method of choice for clinical purposes. However, natural RNA strands are quickly degraded in biological fluids. Chemically synthesized unnatural nucleotides have been developed and introduced into the siRNA strand. For example, modification of the ribose moiety with a 2′-deoxy, 2′-O-methyl, or 2′-fluoro group, or modification of the phosphate backbone have been examined. Although these modifications improve the stability of siRNA in serum, they often cause a decrease in RNAi activity. There is also concern that unnatural RNA derivatives are toxic in the human body. A method to stabilize nontoxic natural RNA strands should be very useful for applications in RNAi technology. We came up with an idea that nano-structural design stabilizes natural RNA. We tested several new designs such as dumbbell RNA, double stranded circular RNA, or branched RNA in biological stability and RNA interference activity. Consequently, dumbbell or branched design offered prolonged RNAi effect due to high biological stability. © 2013 The Pharmaceutical Society of Japan.

DOI： 10.1248/yakushi.12-00239-4

Synthesis of a nucleoside phosphorodithioate analogue responsive to microenvironmental changes through chiral induction 査読有り

Saneyoshi Hisao, Mashimo Takushi, Hatano Ken, Ito Yoshihiro, Abe Hiroshi

TETRAHEDRON LETTERS 54 巻 ( 9 ) 頁： 1080 - 1083 2013年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Tetrahedron Letters

We have synthesized a 2′-aminomethyl branched-chain sugar nucleoside phosphorodithioate from 2,2′-anhydro uridine and subjected the material to a subsequent cyclization reaction under aqueous conditions using bi-functional linkers. The rate of the cyclization reaction was dependent on the leaving group on the bi-functional linkers. The generation of a chiral phosphorous peak from the achiral precursor, as indicated by 31P NMR, was identified as a good indicator for potentially probing the local and global features of the DNA structure. © 2012 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tetlet.2012.12.028

PEGylated Antibodies and DNA in Organic Media and Genetic PEGylation 査読有り

Seiichi Tada, Hiroshi Abe, Yoshihiro Ito

GREEN POLYMER CHEMISTRY: BIOCATALYSIS AND MATERIALS II 1144 巻頁： 223 - 233 2013年

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）出版者・発行元：AMER CHEMICAL SOC

Polyethylene glycol (PEG) is an amphiphilic polymer that is soluble in both water and organic media and can add many functions to biopolymers by conjugation. Here, PEG was coupled to antibodies or oligonucleotides for solubilization into organic media. The former interacted with antigens and the latter formed specific structures and had catalytic activity in organic media. In addition, we attempted to incorporate PEG genetically into a polypeptide. Although the PEG incorporation ratio decreased as the molecular weight of PEG was increased, PEG with a molecular weight of <= 1000 Da was incorporated successfully. The potential applications of PEGylated biopolymers and the methodology for gene PEGylation are discussed.

Fluorogenic probes using 4-substituted-2-nitrobenzenesulfonyl derivatives as caging groups for the analysis of human glutathione transferase catalyzed reactions 査読有り

Shibata Aya, Nakano Yukiko, Ito Mika, Araki Mika, Zhang Jie, Yoshida Yasuhiko, Shuto Satoshi, Mannervik Bengt, Mogenstern Ralf, Ito Yoshihiro, Abe Hiroshi

ANALYST 138 巻 ( 24 ) 頁： 7326 - 7330 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Analyst

We have synthesized a series of 4-substituted-2-nitrobenzene-sulfonyl compounds for caged fluorogenic probes and conducted a Hammett plot analysis using the steady-state kinetic parameters. The results revealed that the glutathione transferase (GST) alpha catalyzed reaction was dependent on the σ value in the same way as the non-enzymatic reaction, whereas the dependence of the σ value of the GST mu and pi was not as pronounced as that of GST alpha. © 2013 The Royal Society of Chemistry.

DOI： 10.1039/c3an01339a

RNA-templated molecule release induced protein expression in bacterial cells 査読有り

Shibata Aya, Ito Yoshihiro, Abe Hiroshi

CHEMICAL COMMUNICATIONS 49 巻 ( 3 ) 頁： 270 - 272 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

We have developed a system to release a biologically active molecule in response to the sequence of a target gene. The releasing system, which was triggered by the reduction of an azidomethyl group, was successfully applied to protein expression induced by the release of IPTG triggered by endogenous RNA in bacterial cells. © 2012 The Royal Society of Chemistry.

DOI： 10.1039/c2cc37826d

One-Pot Ring-Closing Metathesis/1,3-Dipolar Cycloaddition through Assisted Tandem Ruthenium Catalysis: Synthesis of a Dye with Isoindolo[2,1-a]quinoline Structure 査読有り

Arisawa Mitsuhiro, Fujii Yuki, Kato Hiroshige, Fukuda Hayato, Matsumoto Takashi, Ito Mika, Abe Hiroshi, Ito Yoshihiro, Shuto Satoshi

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 52 巻 ( 3 ) 頁： 1003 - 1007 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Angewandte Chemie - International Edition

The one-pot tandem reaction of N-alkyl-N-allyl-2-vinylaniline derivatives with benzo- or naphthoquinones and a ruthenium-alkylidene catalyst leads to isoindolo[2,1-a]quinolines in a variety of colors, which can be altered by exchanging the substituent on the core heterocycle (see scheme). This reaction offers a new synthetic method for π-conjugated small molecules from simple aniline derivatives. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/anie.201206765

Cationic cholesterol-modified gelatin as an in vitro siRNA delivery vehicle 査読有り

Kadengodlu Pallavi Ananda, Aigaki Toshiro, Abe Hiroshi, Ito Yoshihiro

MOLECULAR BIOSYSTEMS 9 巻 ( 5 ) 頁： 965 - 968 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Molecular BioSystems

Polymeric micelle prepared through the self-assembly of cationic cholesterol-modified gelatin was tested for siRNA delivery. It exerted the desired effect of gene knockdown in HeLa cells stably expressing the luciferase gene and achieved a two-fold increase in the knockdown ability when compared to Lipofectamine® 2000. It was found that the polymeric micelle exhibited excellent stability and increased the biological stability of the siRNA in serum. © 2013 The Royal Society of Chemistry.

DOI： 10.1039/c2mb25424g

Rolling Circle Amplification in a Prokaryotic Translation System Using Small Circular RNA 査読有り

Abe Naoko, Hiroshima Michio, Maruyama Hideto, Nakashima Yuko, Nakano Yukiko, Matsuda Akira, Sako Yasushi, Ito Yoshihiro, Abe Hiroshi

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 52 巻 ( 27 ) 頁： 7004 - 7008 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Angewandte Chemie - International Edition

Getting the runaround: Small circular RNA molecules containing an infinite open reading frame were synthesized and tested in an E. coli cell-free translation system. A circular RNA 126 nucleotides in length was found to produce more product than its linear counterpart by two orders of magnitude, because a ribosome can work more effectively towards the elongation on circular RNA than it can on linear RNA in this continuous peptide synthesis. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/anie.201302044

ナノ構造化RNAを用いたRNA干渉法

阿部洋

薬学雑誌 133 巻 ( 3 ) 頁： 373-378 2013年

"PEGylated Antibodies and DNA in Organic Media and Genetic PEGylation" in "Green Polymer 2"

Seiichi Tada, Horoshi Abe, and Yoshihiro Ito

American Chemical Society 頁： 223-233 2013年

PEGylated Antibodies and DNA in Organic Media and Genetic PEGylation

Tada Seiichi, Abe Hiroshi, Ito Yoshihiro

GREEN POLYMER CHEMISTRY: BIOCATALYSIS AND MATERIALS II 1144 巻頁： 223 - 233 2013年

Peroxidase Activity of G-Quadruplex Hemin-Binding DNA Aptamers Determined by Electrochemical Measurement

Kubo I., Eguchi T., Hoshino Y., Liu M., Abe H., Ito T.

BIOENGINEERING BASED ON ELECTROCHEMISTRY 50 巻 ( 28 ) 頁： 1 - 7 2013年

出版者・発行元：ECS Transactions

Certain guanine-rich DNA oligomers that form a G-quadruplex structure and bind hemin inside are called hemin-binding DNA aptamers. A hemin-binding DNA aptamer with the 4c15 sequence forms a parallel G quadraplex and the PS2.M sequence folds and forms an anti-parallel structure. We propose a novel method to examine the peroxidase activity of hemin-binding DNA aptamers through electrochemical measurements. In this study, a polyA chain and thiol were introduced to the 5' terminal of each oligonucleotide. The peroxidase activity of hemin-binding DNA aptamers was determined after modifying a gold electrode with the aptamers. The distance to hemin from the surface of the electrode is most important to determine catalytic activity. Catalytic activity, shown as KM of the examined parallel DNA aptamers, was around 20 μM. 4c15 without a poly A linker showed the highest catalytic activity, and its KM was 19 μM. The KM of the anti-parallel aptamer was almost the same as that of the parallel aptamer. © The Electrochemical Society.

DOI： 10.1149/05028.0001ecst

Immobilization of epidermal growth factor on titanium and stainless steel surfaces via dopamine treatment 査読有り

Kang Jeonghwa, Sakuragi Makoto, Shibata Aya, Abe Hiroshi, Kitajima Takashi, Tada Seiichi, Mizutani Masayoshi, Ohmori Hitoshi, Ayame Hirohito, Son Tae Il, Aigaki Toshiro, Ito Yoshihiro

MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS 32 巻 ( 8 ) 頁： 2552 - 2561 2012年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Materials Science and Engineering C

Titanium and stainless steel were modified with dopamine for the immobilization of biomolecules, epidermal growth factor (EGF). First, the treatment of metal surfaces with a dopamine solution under different pH conditions was investigated. At higher pH, the dopamine solution turned brown and formed precipitates. Treatment of the metals with dopamine at pH 8.5 also resulted in the development of brown color at the surface of the metals. The hydrophobicity of the surfaces increased after treatment with dopamine, independently of pH. X-ray photoelectron spectroscopy revealed the formation of a significant amount of an organic layer on both surfaces at pH 8.5. According to ellipsometry measurements, the organic layer formed at pH 8.5 was about 1000 times as thick as that formed at pH 4.5. The amount of amino groups in the layer formed at pH 8.5 was also higher than that observed in the layer formed at pH 4.5. EGF molecules were immobilized onto the dopamine-treated surfaces via a coupling reaction using carbodiimide. A greater amount of EGF was immobilized on surfaces treated at pH 8.5 compared with pH 4.5. Significantly higher growth of rat fibroblast cells was observed on the two EGF-immobilized surfaces compared with non-immobilized surfaces in the presence of EGF. The present study demonstrated that metals can become bioactive via the surface immobilization of a growth factor and that the effect of the immobilized growth factor on metals was greater than that of soluble growth factor. © 2012 Elsevier B.V.

DOI： 10.1016/j.msec.2012.07.039

Detection of pre-mRNA splicing in vitro by an RNA-templated fluorogenic reaction 査読有り

Tamura Yasutsugu, Furukawa Kazuhiro, Yoshimoto Rei, Kawai Yuto, Yoshida Minoru, Tsuneda Satoshi, Ito Yoshihiro, Abe Hiroshi

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 22 巻 ( 23 ) 頁： 7248 - 7251 2012年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

RNA splicing is an important target for basic research of disease mechanisms and for drug discovery. Here, we report a new method for analysis of the in vitro RNA splicing process that produces fluorescence using a reduction-triggered fluorescence (RETF) probe. The fluorescence signal is produced only when the two probes bind side-by-side with a specific RNA target. Precursor messenger RNA and mature messenger RNA originating from the chicken δ-crystallin (CDC) gene were successfully discriminated in solution using an RETF probe with the assistance of helper oligonucleotide strands. Also, we successfully applied RETF probes to the detection of emerging mature mRNA in an in vitro splicing process. © 2012 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmcl.2012.09.033

ナノ構造を制御したRNA干渉法

阿部洋、阿部奈保子、柴田綾、伊藤嘉浩

アンチセンスDNA/RNA研究会誌 16 巻 ( 2 ) 頁： 3 2012年11月

Design and Synthesis of Indomethacin Analogues That Inhibit P-Glycoprotein and/or Multidrug Resistant Protein without Cox Inhibitory Activity 査読有り

Arisawa Mitsuhiro, Kasaya Yayoi, Obata Tohru, Sasaki Takuma, Nakamura Tomonori, Araki Takuya, Yamamoto Koujirou, Sasaki Akito, Yamano Akihito, Ito Mika, Abe Hiroshi, Ito Yoshihiro, Shuto Satoshi

JOURNAL OF MEDICINAL CHEMISTRY 55 巻 ( 18 ) 頁： 8152 - 8163 2012年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Medicinal Chemistry

We designed and synthesized conformationally restricted analogues and regioisomers of the nonsteroidal anti-inflammatory drug indomethacin. Evaluation of the inhibitory effects of these compounds on COX, P-glycoprotein, and multidrug resistance indicated that NSAIDS modulation of multidrug-resistant P-glycoprotein and multidrug-resistant protein-1 is not associated with COX-1 and COX-2 inhibitory activities. © 2012 American Chemical Society.

DOI： 10.1021/jm301084z

Development of new luminogenic probes for detection of glutathione s-transferase activity in living cells

Ito Mika, Shibata Aya, Abe Hiroshi, Zhang Jie, Morgenstern Ralf, Shuto Satoshi, Ito Yoshihiro

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 244 巻 2012年8月

Development of new luminogenic probes for detection of glutathione s-transferase activity in living cells 査読有り

Mika Ito, Aya Shibata, Hiroshi Abe, Jie Zhang, Ralf Morgenstern, Satoshi Shuto, Yoshihiro Ito

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 244 巻 2012年8月

Universal Caging Group for the in-Cell Detection of Glutathione Transferase Applied to 19F NMR and Bioluminogenic Probes 査読有り

Ito Mika, Shibata Aya, Zhang Jie, Hiroshima Michio, Sako Yasushi, Nakano Yukiko, Kojima-Aikawa Kyoko, Mannervik Bengt, Shuto Satoshi, Ito Yoshihiro, Morgenstern Ralf, Abe Hiroshi

CHEMBIOCHEM 13 巻 ( 10 ) 頁： 1428 - 1432 2012年7月

還元反応を引き金とする off/on 型蛍光プローブを用いたRNA検出法の開発

阿部洋, 柴田綾, 古川和寛, 常田聡, 伊藤嘉浩

化学と生物 50 巻 ( 7 ) 頁： 540 - 544 2012年7月

環元反応を引き金とするoff/on 型蛍光プローブを用いたRNA検出法の開発

阿部洋、柴田綾、古川和寛、常田聡、伊藤嘉浩

化学と生物 50 巻 ( 7月 ) 頁： 540 2012年7月

Oligonucleotide-Templated Reactions for Sensing Nucleic Acids 査読有り

Shibata Aya, Abe Hiroshi, Ito Yoshihiro

MOLECULES 17 巻 ( 3 ) 頁： 2446 - 2463 2012年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Molecules

Oligonucleotide-templated reactions are useful for applying nucleic acid sensing. Various chemistries for oligonucleotide-templated reaction have been reported so far. Major scientific interests are focused on the development of signal amplification systems and signal generation systems. We introduce the recent advances of oligonucleotide-templated reaction in consideration of the above two points. © 2012 by the authors.

DOI： 10.3390/molecules17032446

Synthesis of Dumbbell-Shaped Cyclic RNAs for RNA Interference

Naoko Abe, Hiroshi Abe, Yoshihiro Ito

Current Protocol Nucleic Acid Chemistry 頁： 1-16 2012年

In vitro selection of a photo-responsive peptide aptamer using ribosome display 査読有り

Liu Mingzhe, Tada Seiichi, Ito Mika, Abe Hiroshi, Ito Yoshihiro

CHEMICAL COMMUNICATIONS 48 巻 ( 97 ) 頁： 11871 - 11873 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

A photo-responsive peptide aptamer against microbeads immobilized streptavidin was isolated using in vitroselection combined with photo-manipulation. This is the first example of the introduction of a peptide aptamer in the photo-control of dynamic molecular recognition. © 2012 The Royal Society of Chemistry.

DOI： 10.1039/c2cc36618e

Tuning efficiency of the 4-exo-trig cyclization by the electronic effect: ring closure of 3,3-difluoro-4-pentenyl carbon radicals and synthesis of a gem-difluorocyclobutane nucleoside 査読有り

Kumamoto Hiroki, Kawahigashi Sachiko, Wakabayashi Hiromi, Nakano Tomohiko, Miyaike Tomoko, Kitagawa Yasuyuki, Abe Hiroshi, Ito Mika, Haraguchi Kazuhiro, Balzarini Jan, Baba Masanori, Tanaka Hiromichi

CHEMICAL COMMUNICATIONS 48 巻 ( 89 ) 頁： 10993 - 10995 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

4-exo-trig Cyclization reaction of a 4-pentenyl carbon radical containing the gem-difluoromethylene moiety adjacent to a radical accepting α,β-unsaturated ester was found to proceed efficiently to furnish a novel gem-difluorocyclobutane derivative. The cyclized product could be transformed into a gem-difluoromethylene analogue of oxetanocin T. © 2012 The Royal Society of Chemistry.

DOI： 10.1039/c2cc35876j

Synthesis of dumbbell-shaped cyclic RNAs for RNA interference. 査読有り

Abe N, Abe H, Ito Y

Current protocols in nucleic acid chemistry 1 巻 ( 48 ) 頁： 16.4.11 - 11 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Current Protocols in Nucleic Acid Chemistry

RNA interference (RNAi) is a potent and highly specific gene-silencing phenomenon that was first reported for the nematode Caenorhabditis elegans. It has been discovered that genes could be silenced by introducing double-stranded RNAs (dsRNAs) complementary to the messenger RNA sequences. Since then, RNAi has been shown as an evolutionarily well-conserved process that plays an important role in host defense and in regulation of gene expression. Much effort has been dedicated to the application of the short dsRNA species (short interfering RNAs; siRNAs) as therapeutic agents, as they were shown to be effective in mammalian cells. Recently, we altered the structure of a siRNA molecule and produced dumbbell-shaped nanocircular RNAs. RNA dumbbells were shown to be stabilized in serum compared with its siRNA counterpart, despite their natural RNA strand. It has also been found that RNA dumbbells containing a 23-bp stem and two 9-nt loops exhibit a prolonged RNAi effect in cultured mammalian cells. In this unit, we describe the synthesis of RNA dumbbells from the design, its enzymatic synthesis, and to the purification. © 2012 by John Wiley & Sons, Inc.

DOI： 10.1002/0471142700.nc1604s48

Structure Formation and Catalytic Activity of DNA Dissolved in Organic Solvents 査読有り

Abe Hiroshi, Abe Naoko, Shibata Aya, Ito Keiji, Tanaka Yoshiyuki, Ito Mika, Saneyoshi Hisao, Shuto Satoshi, Ito Yoshihiro

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 51 巻 ( 26 ) 頁： 6475 - 6479 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Angewandte Chemie - International Edition

Equal-opportunity dissolver: By attaching polyethylene glycol at its 5' end, DNA (PEG-DNA) can be solubilized in various organic solvents and was shown to form G-quadruplexes by CD spectroscopy. A complex containing iron(III) protoporphyrin IX (hemin) and G-quadruplex-forming PEG-DNA catalyzed an oxidative reaction in methanol (see scheme). Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/anie.201201111

Positively charged cholesterol-recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions 査読有り

Kadengodlu Pallavi A., Hebishima Takehisa, Takeshima Shin-Nosuke, Ito Mika, Liu Mingzhe, Abe Hiroshi, Aida Yoko, Aigaki Toshiro, Ito Yoshihiro

INTERNATIONAL JOURNAL OF NANOMEDICINE 7 巻頁： 5437 - 5450 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：International Journal of Nanomedicine

Purpose: Recombinant human gelatins with defined molecular weights were modified with cholesterol to make them amphiphilic in nature. We investigated the feasibility of these modified human gelatins acting as a carrier of antigenic proteins for inducing cellular immunity. The aim of this study was to synthesize novel and effective compounds for vaccine delivery in vivo. Methods: Two types of cholesterol-modified gelatin micelles, anionic cholesterol-modified gelatin (aCMG) and cationic-cholesterol modified gelatin (cCMG), were synthesized using different cholesterol derivatives such as the cholesterol-isocyanate (Ch-I) for aCMG and amino-modified cholesterol for cCMG. One was anionic and the other cationic, and therefore they differed in terms of their zeta potential. The aCMG and cCMG were characterized for their size, zeta potential, and in their ability to form micelles. Cytotoxicity was also evaluated. The modified human gelatins were then investigated as a carrier of antigenic proteins for inducing cellular immunity both in vitro in DC 2.4 cells, a murine dendritic cell line, as well as in vivo. The mechanism of entry of the polymeric micelles into the cells was also evaluated. Results: It was found that only cCMG successfully complexed with the model antigenic protein, fluorescein-isothiocyanate ovalbumin (OVA) and efficiently delivered and processed proteins in DC 2.4 cells. It was hypothesized that cCMG enter the cells predominantly by a caveolae-mediated pathway that required tyrosine kinase receptors on the cell surface. Animal testing using mice showed that the cationic cholesterol-modified gelatin complexed with OVA produced significantly high antibody titers against OVA: 2580-fold higher than in mice immunized with free OVA. Conclusion: Conclusively, cCMG has shown to be very effective in stimulating an immune response due to its high efficiency, stability, and negligible cytotoxicity. © 2012 Cárdenas et al, publisher and licensee Dove Medical Press Ltd.

DOI： 10.2147/IJN.S36350

Synthesis, structure, and biological activity of dumbbell-shaped nanocircular RNAs for RNA interference. 査読有り

Abe N, Abe H, Nagai C, Harada M, Hatakeyama H, Harashima H, Ohshiro T, Nishihara M, Furukawa K, Maeda M, Tsuneda S, Ito Y

Bioconjugate chemistry 22 巻 ( 10 ) 頁： 2082 - 92 2011年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioconjugate Chemistry

RNA interference (RNAi) is one of the most promising new approaches for disease therapy. The design of a dumbbell-shaped nanocircular RNA allows it to act as a short interfering RNA (siRNA) precursor. To optimize the design, we studied the relationship between the nanostructure and RNAi activity by synthesizing various RNA dumbbells. An RNA dumbbell with a 23-bp stem and 9-nt loops was the most potent. Sequence analysis by mass spectrometry showed that Dicer could edit RNA dumbbells to siRNA species. The reaction offered the slow release of siRNA species, which conferred prolonged RNAi activity. Introduction of DNA into the loop position significantly stabilized the dumbbell in biological fluid without any loss of RNAi activity. In-depth pharmacological evaluation was performed by introducing dumbbells into HeLa cells that stably express the target luciferase gene. The dumbbells provided a rapid silencing effect and retained this effect for a longer time even at a lower concentration than that at which standard siRNA completely lost RNAi activity. We conclude that an RNA dumbbell with DNA loops is the most promising design for in vivo applications for RNA medicine. © 2011 American Chemical Society.

DOI： 10.1021/bc2003154

Synthesis, Structure, and Biological Activity of Dumbbell-Shaped Nanocircular RNAs for RNA Interference 査読有り

Abe Naoko, Abe Hiroshi, Nagai Chisato, Harada Mitsuru, Hatakeyama Hiroto, Harashima Hideyoshi, Ohshiro Takahito, Nishihara Mizuki, Furukawa Kazuhiro, Maeda Mizuo, Tsuneda Satoshi, Ito Yoshihiro

BIOCONJUGATE CHEMISTRY 22 巻 ( 10 ) 頁： 2082 - 2092 2011年10月

Synthesis and Characterization of a Series of Highly Fluorogenic Substrates for Glutathione Transferases, a General Strategy 査読有り

Zhang Jie, Shibata Aya, Ito Mika, Shuto Satoshi, Ito Yoshihiro, Mannervik Bengt, Abe Hiroshi, Morgenstern Ralf

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 133 巻 ( 35 ) 頁： 14109 - 14119 2011年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of the American Chemical Society

Glutathione transferases (GSTs) are used in biotechnology applications as fusion partners for facile purification and are also overexpressed in certain tumors. Consequently, there is a need for sensitive detection of the enzymes. Here we describe a general strategy for the synthesis and characterization of novel fluorogenic substrates for GSTs. The substrates were synthesized by introducing an electrophilic sulfonamide linkage to fluorescent molecules containing an amino group [e.g., 2,4-dinitrobenzenesulfonamide (DNs) derivatives of coumarin, cresyl violet, and rhodamine]. The derivatives were essentially nonfluorescent, and upon GST catalyzed cleavage of the dinitrobenzenesulfonamide, free fluorophore is released (and 1-glutathionyl-2,4-dinitrobenzene + SO 2). All the coumarin-, cresyl violet- and rhodamine-based fluorogenic probes turned out to be good substrates for most GSTs, especially for GSTA 1-1, in terms of strong fluorescence increases (71-1200-fold), high k cat/K m values (10 4-10 7 M -1 s -1) and significant rate enhancements (10 6-10 9-fold). The substrates were successfully applied to quantitate very low levels of GST activity in cell extracts and DNs-cresyl violet was also successfully applied to the imaging of microsomal MGST 1 activity in living cells. The cresyl violet stained cells retained their fluorescence after fixation, which is a very useful property. In summary, we describe a general and versatile strategy to generate fluorogenic GST substrates, some of them providing the most sensitive assays so far described for GSTs. © 2011 American Chemical Society.

DOI： 10.1021/ja205500y

Synthesis of 4 '-Ethynyl-2 '-deoxy-4 '-thioribonucleosides and Discovery of a Highly Potent and Less Toxic NRTI 査読有り

Haraguchi Kazuhiro, Shimada Hisashi, Kimura Keigo, Akutsu Genta, Tanaka Hiromichi, Abe Hiroshi, Hamasaki Takayuki, Baba Masanori, Gullen Elizabeth A., Dutschman Ginger E., Cheng Yung-Chi, Balzarini Jan

ACS MEDICINAL CHEMISTRY LETTERS 2 巻 ( 9 ) 頁： 692 - 697 2011年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACS Medicinal Chemistry Letters

The synthesis of 4′-ethynyl-2′-deoxy-4′- thioribonucleosides was carried out utilizing an electrophilic glycosidation in which 4-ethynyl-4-thiofuranoid glycal 16 served as a glycosyl donor. Electrophilic glycosidation between 16 and the silylated nucleobases (N 4-acetylcytosine, N 6-benzoyladenine, and N 2-acetyl-O 6-diphenylcarbamoylguanine) was carried out in the presence of N-iodosuccinimide (NIS), leading to the exclusive formation of the desired β-anomers 29, 33, and 36. Anti-HIV studies demonstrated that these 4′-thio nucleosides were less cytotoxic to T-lymphocyte (i.e., MT-4 cells) than the corresponding 4′-ethynyl derivatives of 2′-deoxycytidine (44), 2′-deoxyadenosine (45), and 2′-deoxyguanosine (46). Comparison of the selectivity indices (SI) was made between 4′-thionucleosides (32, 41, and 43) and the corresponding 4′-oxygen analogues 44-46 by using the reported CC 50 and EC 50 values. In the case of cytosine and adenine nucleosides, comparable SI values were obtained as follows: 32 (545) and 44 (458); 41 (>230) and 45 (1630). In contrast, 4′-ethynyl-2′-deoxy-4′- thioguanosine 43 was found to possess a SI value of >18200, which is 20 times better than that of 46 (933). © 2011 American Chemical Society.

DOI： 10.1021/ml2001054

PEGylated antibody in organic media 査読有り

Liu Mingzhe, Xu Muye, Loh Xian Jun, Abe Hiroshi, Tsumuraya Takeshi, Fujii Ikuo, Li Jun, Son Tae Il, Ito Yoshihiro

JOURNAL OF BIOSCIENCE AND BIOENGINEERING 111 巻 ( 5 ) 頁： 564 - 568 2011年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Bioscience and Bioengineering

Antibodies were covalently conjugated with poly(ethylene glycol) (PEG) and the properties of the PEGylated antibodies in organic media were investigated. Two types of monoclonal antibody were used in this study. One was a monoclonal antibody (abzyme) that was prepared against a hapten mimicking a transition state of hydrolysis. Another was a monoclonal antibody against estrogen, which is not soluble in water. By electrophoresis and mass spectral analysis, the covalent conjugation with PEG chains was confirmed. The PEGylated antibodies bound to antigens and the PEGylated abzyme catalyzed a hydrolysis reaction in an aqueous solution. The PEGylated antibodies were soluble in dichloromethane and acetone and interacted with antigen either in dichloromethane or in acetone. In conclusion, PEGylated antibodies can be employed as analytical tools for water-insoluble analytes. © 2011 The Society for Biotechnology, Japan.

DOI： 10.1016/j.jbiosc.2011.01.001

Indomethacin Analogues that Enhance Doxorubicin Cytotoxicity in Multidrug Resistant Cells without Cox Inhibitory Activity 査読有り

Arisawa Mitsuhiro, Kasaya Yayoi, Obata Tohru, Sasaki Takuma, Ito Mika, Abe Hiroshi, Ito Yoshihiro, Yamano Akihito, Shutot Satoshi

ACS MEDICINAL CHEMISTRY LETTERS 2 巻 ( 5 ) 頁： 353 - 357 2011年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACS Medicinal Chemistry Letters

Conformationally restricted indomethacin analogues were designed and prepared from the corresponding 2-substituted indoles, which were synthesized by a one-pot isomerization/enamide-ene metathesis as the key reaction. Conformational analysis by calculations, NMR studies, and X-ray crystallography suggested that these analogues were conformationally restricted in the s-cis or the s-trans form due to the 2-substituent as expected. Their biological activities on cyclooxygenase-1 (COX-1) inhibition, cyclooxygenase-2 (COX-2) inhibition, and modulation of MRP-1-mediated multidrug resistance (MDR) are described. Some of these indomethacin analogues enhanced doxorubicin cytotoxicity, although they do not have any COX inhibitory activity, which suggests that the MDR-modulating effect of an NSAID can be unassociated with its COX-inhibitory activity. This may be an entry into the combination chemotherapy of doxorubicin with a MDR modulator. © 2011 American Chemical Society.

DOI： 10.1021/ml100292y

Creation of Polymer Catalysts by Molecular Evolutionary Engineering and Hybridization of Biocatalysts 査読有り

Mingzhe Liu, Hiroshi Abe, Yoshihiro Ito

KOBUNSHI RONBUNSHU 68 巻 ( 7 ) 頁： 405 - 416 2011年

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：SOC POLYMER SCIENCE JAPAN

We developed a new type of oligonucleotide-based catalysts which can be called DNAzyme, ribozyme, or aptazyme. They were in vitro selected from a random sequence library of DNA or RNA for binding hemin. The selected oligonucleotides exhibited peroxidase activity by forming a complex with hemin. In addition, an azobenzene-containing oligonucleotide which bound to hemin was also selected. The binding and the catalytic activity was controlled by photo-irradiation. Subsequently, hybridized biocatalysts with synthetic polymers were synthesized. Polyethylene glycol-conjugated antibodies and oligonucleotides were synthesized and it was demonstrated that they were active in organic media. A photo-responsive polymer was also conjugated with an enzyme and its catalytic activity was controlled in organic media.

Fluorescence Detection of Intron Lariat RNA with Reduction-Triggered Fluorescent Probes 査読有り

Furukawa Kazuhiro, Abe Hiroshi, Tamura Yasutsugu, Yoshimoto Rei, Yoshida Minoru, Tsuneda Satoshi, Ito Yoshihiro

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 50 巻 ( 50 ) 頁： 12020 - 12023 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Angewandte Chemie - International Edition

Fluorescence through proximity: Reduction-triggered fluorescent probes bearing either a fluorescein derivative or triphenylphosphine bind sequence-specifically at the branch site of lariat RNA. The chemical reaction between the two substrates leads to the formation of unquenched fluorescein and the emission of a fluorescence signal (see picture). Prelariat and lariat structures can be distinguished by using this method. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/anie.201104425

Synthesis and characterization of small circular double-stranded RNAs 査読有り

Abe Naoko, Abe Hiroshi, Ohshiro Takahito, Nakashima Yuko, Maeda Mizuo, Ito Yoshihiro

CHEMICAL COMMUNICATIONS 47 巻 ( 7 ) 頁： 2125 - 2127 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

Small circular double-stranded RNAs (dsRNAs) were synthesized. The structural analysis by atomic force microscopy gave direct images for the interpretation of the structural strain present in circular dsRNAs. Finally, we demonstrated that circular dsRNA caused RNAi effect in cells. © 2011 The Royal Society of Chemistry.

DOI： 10.1039/c0cc04551a

Branched RNA nanostructures for RNA interference 査読有り

Nakashima Yuko, Abe Hiroshi, Abe Naoko, Aikawa Kyoko, Ito Yoshihiro

CHEMICAL COMMUNICATIONS 47 巻 ( 29 ) 頁： 8367 - 8369 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

Branched RNAs with three- or four-way junctions were designed by assembling single-stranded RNA for RNA interference. Human Dicer transformed branched RNAs into about 20 base pairs of double-stranded RNA, which is a standard siRNA species. Our tetramer design provides a potent silencing effect over a period of 5 days. © 2011 The Royal Society of Chemistry.

DOI： 10.1039/c1cc11780g

Characterization of New Potential Anticancer Drugs Designed To Overcome Glutathione Transferase Mediated Resistance 査読有り

Johansson Katarina, Ito Mika, Schophuizen Carolien M. S., Thengumtharayil Sherin Mathew, Heuser Vanina D., Zhang Jie, Shimoji Miyuki, Vahter Marie, Wee Han Ang, Dyson Paul J., Shibata Aya, Shuto Satoshi, Yoshihiro Ito, Abe Hiroshi, Morgenstern Ralf

MOLECULAR PHARMACEUTICS 8 巻 ( 5 ) 頁： 1698 - 1708 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Molecular Pharmaceutics

Resistance against anticancer drugs remains a serious obstacle in cancer treatment. Here we used novel strategies to target microsomal glutathione transferase 1 (MGST1) and glutathione transferase pi (GSTP) that are often overexpressed in tumors and confer resistance against a number of cytostatic drugs, including cisplatin and doxorubicin (DOX). By synthetically combining cisplatin with a GST inhibitor, ethacrynic acid, to form ethacraplatin, it was previously shown that cytosolic GST inhibition was improved and that cells became more sensitive to cisplatin. Here we show that ethacraplatin is easily taken up by the cells and can reverse cisplatin resistance in MGST1 overexpressing MCF7 cells. A second and novel strategy to overcome GST mediated resistance involves using GST releasable cytostatic drugs. Here we synthesized two derivatives of DOX, 2,4-dinitrobenzenesulfonyl doxorubicin (DNS-DOX) and 4-mononitrobenzenesulfonyl doxorubicin (MNS-DOX) and showed that they are substrates for MGST1 and GSTP (releasing DOX). MGST1 overexpressing cells are resistant to DOX. The resistance is partially reversed by DNS-DOX. Interestingly, the less reactive MNS-DOX was more cytotoxic to cells overexpressing MGST1 than control cells. It would appear that, by controlling the reactivity of the prodrug, and thereby the DOX release rate, selective toxicity to MGST1 overexpressing cells can be achieved. In the case of V79 cells, DOX resistance proportional to GSTP expression levels was noted. In this case, not only was drug resistance eliminated by DNS-DOX but a striking GSTP-dependent increase in toxicity was observed in the clonogenic assay. In summary, MGST1 and GSTP resistance to cytostatic drugs can be overcome and cytotoxicity can be enhanced in GST overexpressing cells. © 2011 American Chemical Society.

DOI： 10.1021/mp2000692

進化分子工学による高分子触媒合成と生体高分子のハイブリット化

劉明哲、阿部洋、伊藤嘉浩

高分子論文集 68 巻 ( 7 ) 頁： 405-416 2011年

Creation of Polymer Catalysts by Molecular Evolutionary Engineering and Hybridization of Biocatalysts

Liu Mingzhe, Abe Hiroshi, Ito Yoshihiro

KOBUNSHI RONBUNSHU 68 巻 ( 7 ) 頁： 405 - 416 2011年

出版者・発行元：Kobunshi Ronbunshu

We developed a new type of oligonucleotide-based catalysts which can be called DNAzyme, ribozyme, or aptazyme. They were in vitro selected from a random sequence library of DNA or RNA for binding hemin. The selected oligonucleotides exhibited peroxidase activity by forming a complex with hemin. In addition, an azobenzene-containing oligonucleotide which bound to hemin was also selected. The binding and the catalytic activity was controlled by photo-irradiation. Subsequently, hybridized biocatalysts with synthetic polymers were synthesized. Polyethylene glycol-conjugated antibodies and oligonucleotides were synthesized and it was demonstrated that they were active in organic media. A photo-responsive polymer was also conjugated with an enzyme and its catalytic activity was controlled in organic media. © 2011, The Society of Polymer Science Japan.

DOI： 10.1295/koron.68.405

Synthesis and properties of a novel nucleoside derivative possessing a 2,3,5,6-tetraazabenzo[cd]azulene skeleton 査読有り

Hirama Yasuyuki, Abe Hiroshi, Minakawa Noriaki, Matsuda Akira

TETRAHEDRON 66 巻 ( 43 ) 頁： 8402 - 8406 2010年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Tetrahedron

We describe herein the synthesis and properties of the novel nucleoside derivative, 4,7-diamino-2-(2-deoxy-β-d-erythro-pentofuranosyl)-2,6-dihydro- 7H-2,3,5,6-tetraazabenzo[cd]azulene (1). The palladium catalyzed cross-coupling reaction of 2,4-diamino-5-iodo-7-(2-deoxy-β-d-erythro-pentofuranosyl) pyrrolo[2,3-d]pyrimidine (9) with acrylonitrile afforded 2,4-diamino-5-[(E)-1- cyano-2-ethenyl]-7-(2-deoxy-β-d-erythro-pentofuranosyl)pyrrolo[2,3-d] pyrimidine (10) in 77% yield, which was treated with NaOMe in MeOH in the presence of NaSPh to give the desired 1 in 64% yield. Whereas 1 was stable in concentrated ammonia at room temperature, it was gradually hydrolyzed in water to give 4-amino-2-(2-deoxy-β-d-erythro-pentofuranosyl)-2,6-dihydro-7H-2,3, 5,6-tetraazabenzo[cd]azulen-7-one (12). Density functional calculations indicated that 12 was 20 kcal/mol more thermodynamically stable than 1 in a model study. © 2010 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tet.2010.08.063

Hemin-binding aptamers and aptazymes

Liu M., Abe H., Ito Y.

ACS Symposium Series 1043 巻頁： 111 - 123 2010年8月

出版者・発行元：ACS Symposium Series

Aptamers are oligonucleotides that have been engineered through repeated rounds of in vitro selection or by the process of Systematic Evolution of Ligands by EXponential enrichment (SELEX) to bind to various molecular targets such as small molecules, proteins, nucleic acids, and even cells and tissues. Aptamers offer similar molecular recognition properties to the more commonly used antibodies. In addition to their ability to discriminate their target, aptamers offer some advantages over antibodies. For example, aptamers can be engineered completely in a test tube and are readily produced using chemical processes. On the other hand, some aptamers that have been developed exhibit enzyme activity after binding to their target molecule, and function as 'aptazymes'. Numerous aptamers and aptazymes have been used in biotechnology, diagnostics and therapy. In this study, we demonstrate that natural DNA/RNA aptamers produced by selection in vitro, not only bind to hemin, but also show peroxidase activity when in such complexes (i.e., they function as aptazymes). © 2010 American Chemical Society.

DOI： 10.1021/bk-2010-1043.ch009

In vitro selection of a photoresponsive RNA aptamer to hemin 査読有り

Liu Mingzhe, Jinmei Hiroshi, Abe Hiroshi, Ito Yoshihiro

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 20 巻 ( 9 ) 頁： 2964 - 2967 2010年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

A photoresponsive RNA aptamer to hemin was selected in vitro from a random sequence library of RNAs with azobenzene residues. The aptamer bound to hemin under visible light irradiation and was released by ultraviolet light. © 2010 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmcl.2010.02.109

New fluorogenic probe for detection of glutathione s-transferase activity in living cells

Ito Mika, Abe Hiroshi, Shibata Aya, Shimizu Shigeru, Alander Johan, Morgenstern Ralf, Ito Yoshihiro

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 239 巻 2010年3月

Design and synthesis of indole derivatives of adenophostin A. A entry into subtype-selective IP3 receptor ligands 査読有り

Mochizuki Tetsuya, Tanimura Akihiko, Nezu Akihiro, Ito Mika, Abe Hiroshi, Ito Yoshihiro, Arisawa Mitsuhiro, Shuto Satoshi

TETRAHEDRON LETTERS 51 巻 ( 6 ) 頁： 977 - 979 2010年2月

掲載種別：研究論文（学術雑誌）出版者・発行元：Tetrahedron Letters

Indole derivatives 3a and 3b of adenophostin A (2) in which the adenine of 2 was replaced with indole or 4-fluoroindole was designed as potential inositol trisphosphate receptor ligands. These target compounds were successfully synthesized from the key disaccharide unit 6. Biological evaluation showed that 3b selectively activates IP3R1, a subtype of IP3 receptors. © 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tetlet.2009.12.045

Synthesis and structural and pharmacological properties of cyclopropane-based conformationally restricted analogs of 4-methylhistamine as histamine H-3/H-4 receptor ligands 査読有り

Kobayashi Takaaki, Watanabe Mizuki, Yoshida Akira, Yamada Shizuo, Ito Mika, Abe Hiroshi, Ito Yoshihiro, Arisawa Mituhiro, Shuto Satoshi

BIOORGANIC & MEDICINAL CHEMISTRY 18 巻 ( 3 ) 頁： 1076 - 1082 2010年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry

On the basis of the previous results on a histamine H4 receptor agonist 4-methylhistamine and a cyclopropane-based conformationally restricted analog CEIC (3) with potent H3/H4 receptor antagonistic effect, 4-methylhistamine analogs 4 and 5 of CEIC were designed and synthesized. Compound 4 showed strong affinity (Ki = 38.7 nM) for the H3 receptor, which was more potent than a well-known H3 antagonist thioperamide. Stable tautomer and conformation of 3 and 4, which can affect the pharmacological activity, were analyzed by ab initio calculations. © 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmc.2009.12.046

Hemin-Binding Aptamers and Aptazymes 査読有り

Mingzhe Liu, Hiroshi Abe, Yoshihiro Ito

GREEN POLYMER CHEMISTRY: BIOCATALYSIS AND BIOMATERIALS 1043 巻頁： 111 - 123 2010年

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）出版者・発行元：AMER CHEMICAL SOC

Aptamers are oligonucleotides that have been engineered through repeated rounds of in vitro selection or by the process of Systematic Evolution of Ligands by EXponential enrichment (SELEX) to bind to various molecular targets such as small molecules, proteins, nucleic acids, and even cells and tissues. Aptamers offer similar molecular recognition properties to the more commonly used antibodies. In addition to their ability to discriminate their target, aptamers offer some advantages over antibodies. For example, aptamers can be engineered completely in a test tube and are readily produced using chemical processes. On the other hand, some aptamers that have been developed exhibit enzyme activity after binding to their target molecule, and function as 'aptazymes'. Numerous aptamers and aptazymes have been used in biotechnology, diagnostics and therapy. In this study, we demonstrate that natural DNA/RNA aptamers produced by selection in vitro, not only bind to hemin, but also show peroxidase activity when in such complexes (i.e., they function as aptazymes).

Photoactivatable fluorescein derivatives with azidomethyl caging groups for tracing oligonucleotides in living human cells 査読有り

Furukawa Kazuhiro, Abe Hiroshi, Tsuneda Satoshi, Ito Yoshihiro

ORGANIC & BIOMOLECULAR CHEMISTRY 8 巻 ( 10 ) 頁： 2309 - 2311 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Organic and Biomolecular Chemistry

A new photocaged fluorescent compound, azidomethyl fluorescein, was successfully utilized to monitor the dynamics of oligonucleotides in living human cells. © 2010 The Royal Society of Chemistry.

DOI： 10.1039/b926100a

Hemin-Binding Aptamers and Aptazymes

Mingahe Liu, Horoshi Abe, Yoshihiro Ito

ACS Symposium Series 1043(Green Polymer Chemistry) 頁： 111-123 2010年

Hemin-Binding Aptamers and Aptazymes

Liu Mingzhe, Abe Hiroshi, Ito Yoshihiro

GREEN POLYMER CHEMISTRY: BIOCATALYSIS AND BIOMATERIALS 1043 巻頁： 111 - 123 2010年

Establishment of a deformable aorta model based on patient's chest CT data

Tokuyasu T., Shuto T., Yufu K., Abe H., Marui A., Kanao S., Komeda M.

2009 4th International Conference on Innovative Computing, Information and Control, ICICIC 2009 頁： 1339 - 1342 2009年12月

出版者・発行元：2009 4th International Conference on Innovative Computing, Information and Control, ICICIC 2009

For arch aorta surgery, cardiac surgeons approach to the diseased area with median incision and stemotomy according to the operation plan based on patient's medical images. Even using latest medical diagnosis instruments, however, it is impossible to estimate the shape of aneurysm, so that operation planning plan based on medical images is hard to be perfect. As the mentioned above, cardiac surgeons can not recognize the statement of patient's aneurysm before operation. Because of difficulty to completely recognize the statement of aneurysm from patient's medical images, operation plan often has to be changed during the operation. Therefore the coauthor cardiac surgeons highly expect a diagnosis support system that enables them to observe the shape of aneurysm from any angle and to virtually palpate it in order to make more safety operation plan. Then this study aims to develop of a diagnosis assistant system by using virtual reality and haptic interface under considering realtime interaction between them. © 2009 IEEE.

DOI： 10.1109/ICICIC.2009.194

Reduction-Triggered Fluorescence Probe for Peptide-Templated Reactions 査読有り

Shibata Aya, Abe Hiroshi, Furukawa Kazuhiro, Tsuneda Satoshi, Ito Yoshihiro

CHEMICAL & PHARMACEUTICAL BULLETIN 57 巻 ( 11 ) 頁： 1223 - 1226 2009年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical and Pharmaceutical Bulletin

We developed a new nucleic acid-based fluorescence probe for protein detection. The method is based on the scission of an aptamer into two probes, which are then attached with a chemically reactive fluorogenic compound. The protein-dependent association of the two probes accelerates a reduction-triggered fluorogenic reaction and indicates the presence of the target protein, which is detected using a fluorescence readout. The fluorescence signal is generated via the deprotection of the azidomethyl group of fluorescein. The arginine-rich motif peptide of the human immunodeficiency virus-1 Rev protein was targeted by this type of probe. Emission was detected at 522 nm and was enhanced by about 19.4-fold in the presence of the target peptide. An oligonucleotide-based reduction-triggered fluorescence probe was successfully applied to the detection of the Rev peptide in solution. © 2009 Pharmaceutical Society of Japan.

DOI： 10.1248/cpb.57.1223

Characterization of a new fluorogenic substrate for microsomal glutathione transferase 1 査読有り

Alander Johan, Johansson Katarina, Heuser Vanina Dahlstrom, Farebo Henny, Jarvliden Julia, Abe Hiroshi, Shibata Aya, Ito Mika, Ito Yoshihiro, Morgenstern Ralf

ANALYTICAL BIOCHEMISTRY 390 巻 ( 1 ) 頁： 52 - 56 2009年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Analytical Biochemistry

A new thiol-reactive electrophilic, disubstituted rhodamine-based fluorogenic probe (bis-2,4-dinitrobenzenesulfonyl rhodamine [BDR]) with very high quantum yield was synthesized and described recently [A. Shibata et al., Bioorg. Med. Chem. Lett. 18 (2008) 2246-2249]. Because hydrophobic electrophiles are often conjugated by glutathione transferases, the BDR or monosubstituted rhodamine derivatives (2,4-dinitrobenzenesulfonyl rhodamine [DR]) were tested with microsomal glutathione transferase 1 (MGST1) and shown to function as substrates. The kinetic parameters for purified enzyme and DR were kcat = 0.075 ± 0.005 s-1 and Km = 21 ± 3 μM (kcat/Km = 3.6 × 103 ± 5.6 × 102 M-1 s-1), giving a rate enhancement of 106 compared with the nonenzymatic reaction. In cells overexpressing MGST1, the addition of BDR caused a time-dependent increase of fluorescence compared with control cells. Preincubating the cells with a thiol reagent (N-ethylmaleimide) abolished the fluorescent signal. By using DR, we could determine the MGST1 activity in whole cell extracts with high sensitivity. In addition, the activity could be increased by thiol reagents (a hallmark of MGST1). Thus, we have identified a new fluorogenic substrate for MGST1 that will be a useful tool in the study of this enzyme and related enzymes. © 2009 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ab.2009.03.046

Reduction-Triggered Fluorescent Amplification Probe for the Detection of Endogenous RNAs in Living Human Cells 査読有り

Furukawa Kazuhiro, Abe Hiroshi, Hibino Kayo, Sako Yasushi, Tsuneda Satoshi, Ito Yoshihiro

BIOCONJUGATE CHEMISTRY 20 巻 ( 5 ) 頁： 1026 - 1036 2009年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioconjugate Chemistry

Oligonucleotide-templated reactions are attracting attention as a method for RNA detection in living cells. Previously, a reduction-triggered fluorescence probe has been reported that is based on azide reduction to switch fluorescence on. In this article, we report a more advanced probe, a reduction-triggered fluorescent amplification probe that is capable of amplifying a target signal. Azidomethyl fluorescein was newly synthesized and introduced into a probe. Azido-masked fluorescein on the probe showed a strong turn-on fluorescence signal upon oligonucleotide-templated Staudinger reduction. The catalytic reaction of the probe offered a turnover number of 50 as fluorescence readout within 4 h. Finally, probes were introduced into human leukemia HL-60 cells by use of streptolysin O pore-forming peptide. We successfully detected and quantitated the 28S rRNA and β-Actin mRNA signal above the background by flow cytometry. In addition, the same RNA targets were imaged by fluorescence microscopy. The data suggest that a reduction-triggered amplification probe may be a powerful tool in analyzing the localization, transcription, or processing of RNA species in living eukaryotic cells. © 2009 American Chemical Society.

DOI： 10.1021/bc900040t

In vitro selection of hemin-binding catalytic RNA 査読有り

Liu Mingzhe, Kagahara Takuma, Abe Hiroshi, Ito Yoshihiro

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 19 巻 ( 5 ) 頁： 1484 - 1487 2009年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

Catalytic RNAs with peroxidase activity were obtained by the in vitro selection of RNA aptamer-binding hemin. One of the RNA aptamers selected showed binding affinity to hemin with a dissociation constant of 0.8 μM and exhibited high peroxidase activity by forming a complex with hemin. The catalytic efficiency of the RNA-hemin complex was 10-fold higher than that of hemin alone. © 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmcl.2009.01.016

細胞機能制御高分子と再生医療高分子

伊藤嘉浩、北嶋隆、阿部洋

細胞機能制御高分子と再生医療高分子 58 巻 ( 3月 ) 頁：全 2009年3月

Direct In Vitro Selection of Hemin-Binding DNA Aptamer with Peroxidase Activity 査読有り

Liu Mingzhe, Kagahara Takuma, Abe Hiroshi, Ito Yoshihiro

BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN 82 巻 ( 1 ) 頁： 99 - 104 2009年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bulletin of the Chemical Society of Japan

A novel hemin-binding DNA was synthesized using in vitro selection (SELEX) method. The selected DNA not only bound to the hemin, but also had peroxidase activity when complexed with hemin. A pool of 104-nt single-stranded DNA (ssDNA) molecules containing a randomized sequence of 60nt was synthesized. The DNA pool having random sequences was incubated with a hemin-immobilized affinity column. Bound DNAs were eluted with hemin solution and amplified by PCR with biotin-labeled primers. ssDNAs were isolated from the biotin-labeled double-stranded DNA (dsDNA) molecules after each selection process. After four rounds of selection process, the selected DNAs were cloned and sequenced. Four DNA aptamers were chemically synthesized from the sequenced clones. Two aptamers exhibited binding affinity to hemin and peroxidase activity. A 21-nt oligonucleotide was designed from the aptamer sequence that formed a complex with hemin and exhibited high peroxidase activity. © 2009 The Chemical Society of Japan.

DOI： 10.1246/bcsj.82.99

Protein detection using oligonucleotide probes 査読有り

Aya Shibata, Hiroshi Abe, Kazuhiro Furukawa, Satoshi Tsuneda, Yoshihiro Ito

Nucleic Acids Symposium Series 53 巻頁： 157 - 158 2009年

DNA templated nucleophilic aromatic substitution reactions for fluorogenic sensing of oligonucleotides 査読有り

Shibata Aya, Abe Hiroshi, Ito Mika, Kondo Yuko, Shimizu Shigeru, Aikawa Kyoko, Ito Yoshihiro

CHEMICAL COMMUNICATIONS 43 巻 ( 43 ) 頁： 6586 - 6588 2009年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemical Communications

We have developed an SNAr reaction-triggered fluorescence probe using a new fluorogenic compound derivatized from 7-aminocoumarin for oligonucleotides detection. © The Royal Society of Chemistry 2009.

DOI： 10.1039/b912896d

Reduction-triggered red fluorescent probes for dual-color detection of oligonucleotide sequences 査読有り

Furukawa Kazuhiro, Abe Hiroshi, Wang Jin, Uda Miwako, Koshino Hiroyuki, Tsuneda Satoshi, Ito Yoshihiro

ORGANIC & BIOMOLECULAR CHEMISTRY 7 巻 ( 4 ) 頁： 671 - 677 2009年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Organic and Biomolecular Chemistry

We have developed a new red fluorogenic compound derived from naphthorhodamine for a reduction-triggered fluorescence probe to sense oligonucleotides. The fluorogenic reaction between naphthorhodamine azide derivatives and reducing reagents such as triphenylphosphine (TPP) on the DNA target does not use any enzyme or reagent, and fluoresces at 650 nm. The probes were used for dual color detection of a single nucleotide difference on the leukemia-related bcr/abl gene. © 2009 The Royal Society of Chemistry.

DOI： 10.1039/b817228e

Synthetic nanocircular RNA for controlling of gene expression 査読有り

Hiroshi Abe, Naoko Abe, Miwako Uda, Satoshi Tsuneda, Yoshihiro Ito

Nucleic Acids Symposium Series 53 巻頁： 65 - 66 2009年

Synthetic nanocircular RNA for controlling of gene expression. 査読有り

Abe H, Abe N, Uda M, Tsuneda S, Ito Y

Nucleic acids symposium series (2004) ( 53 ) 頁： 65 - 6 2009年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic acids symposium series (2004)

We designed and synthesized dumbbell-shaped nanocircular RNAs for RNA interference applications, which consist of a stem and two loops(1). RNA dumbbells are specifically recognized and cleaved by the human Dicer enzyme, and are thus transformed into double-stranded RNA in cells, although this RNA is resistant to degradation in serum. The structure was optimized to maximize its RNAi activity. The most potent activity was achieved when the stem length was 23 base pairs. The RNAi activity is prolonged by the shape of the molecule, an endless structure, compared with that of normal siRNA.

DOI： 10.1093/nass/nrp033

Protein detection using oligonucleotide probes. 査読有り

Shibata A, Abe H, Furukawa K, Tsuneda S, Ito Y

Nucleic acids symposium series (2004) ( 53 ) 頁： 157 - 8 2009年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic acids symposium series (2004)

We developed a new nucleic acid-based fluorescence probe for protein detection. The method is based on the scission of an aptamer into two probes, which are then attached with a chemically reactive fluorogenic compound. The protein-dependent association of the two probes accelerates a chemical reaction and indicates the presence of the target protein, which is detected using a fluorescence readout. The arginine-rich motif peptide was targeted by this type of probe. In presence of the peptide, the fluorescence signal at 450 nm increased, and no significant increase in fluorescence was observed in the absence of the peptide. An oligonucleotide-based fluorescence probe was successfully applied to the detection of the ARM peptide in solution.

DOI： 10.1093/nass/nrp079

In vitro selection of RNA aptamer to hemin.

Liu M., Kagahara T., Abe H., Ito Y.

Nucleic acids symposium series (2004) ( 52 ) 頁： 513 - 514 2008年12月

出版者・発行元：Nucleic acids symposium series (2004)

A new RNA aptamer-binding hemin was synthesized by the in vitro selection (SELEX) method. A pool of 103 bases single strand DNAs containing a randomized sequence of 59 bases was synthesized. The pool was incubated with hemin on hemin-immobilized affinity column. Bound RNAs were eluted off with hemin solution and amplified by PCR. After 3 rounds of selection process, the selected RNAs were cloned and sequenced. Some RNA aptamers, which have affinity to hemin, was selected.

Nanocircular RNAs for RNA interference.

Abe H., Abe N., Harada M., Tsuneda S., Ito Y.

Nucleic acids symposium series (2004) ( 52 ) 頁： 505 - 506 2008年12月

出版者・発行元：Nucleic acids symposium series (2004)

We designed and synthesized dumbbell-shaped nanocircular RNAs for RNA interference applications, which consist of a stem and two loops(1). RNA dumbbells are specifically recognized and cleaved by the human Dicer enzyme, and are thus transformed into double-stranded RNA in cells, although this RNA is resistant to degradation in serum. The structure was optimized to maximize its RNAi activity. The most potent activity was achieved when the stem length was 23 base pairs. The RNAi activity is prolonged by the shape of the molecule, an endless structure, compared with that of normal siRNA.

Fluorogenic probe triggered by reduction for nucleic acids sensing.

Furukawa K., Abe H., Wang J., Oki K., Uda M., Tsuneda S., Ito Y.

Nucleic acids symposium series (2004) ( 52 ) 頁： 353 - 354 2008年12月

出版者・発行元：Nucleic acids symposium series (2004)

A reduction-triggered fluorescence probe with a new fluorogenic compound derivatized from rhodamine 110 was developed for sensing oligonucleotides. The chemistry to activate the compound involves the reaction between the azide group of rhodamine derivatives and reducing reagents, with the fluorescence signal appearing after reduction of the azide group. The reaction proceeds under biological conditions to produce fluorescence signal within 10-20 min in the presence of target DNA or RNA. The probes were successfully applied to the detection of oligonucleotides in solution and endogenous RNA in bacterial cells.

Fluorescence generation from tandem repeats of a malachite green RNA aptamer using rolling circle transcription 査読有り

Furukawa Kazuhiro, Abe Hiroshi, Abe Naoko, Harada Mitsuru, Tsuneda Satoshi, Ito Yoshihiro

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 18 巻 ( 16 ) 頁： 4562 - 4565 2008年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

We demonstrate a generation of tandem repeats of a malachite green (MG) RNA aptamer using rolling circle transcription. To keep the higher-order structure of each aptamer on long RNA, we designed a sequence of circular DNA with a 14-base linker. T7 RNA polymerase was superior to Escherichia coli RNA polymerase in the specific transcription of the MG RNA aptamer. Finally, the generation of the fluorescence signal was confirmed from aptamer repeats with MG. © 2008 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmcl.2008.07.040

A reduction-triggered fluorescence probe for sensing nucleic acids 査読有り

Abe Hiroshi, Wang Jin, Furukawa Kazuhiro, Oki Kazuma, Uda Miwako, Tsuneda Satoshi, Ito Yoshihiro

BIOCONJUGATE CHEMISTRY 19 巻 ( 6 ) 頁： 1219 - 1226 2008年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioconjugate Chemistry

We have developed a reduction-triggered fluorescence probe with a new fluorogenic compound derivatized from Rhodamine for sensing oligonucleotides. The chemistry to activate the compound involves the reaction between the azide group of rhodamine derivatives and the reducing reagents, with the fluorescence signal appearing after reduction of the azide group. The signal/background ratio of this fluorogenic compound reached 2100-fold enhancement in fluorescence intensity. Dithio-1,4-threitol or triphenylphosphine as reducing reagents were successfully utilized for this chemistry to be introduced into the DNA probe. The genetic detection requires that two strands of DNA bind onto target oligonucleotides, one probe carrying a reducible fluorogenic compound while the other carries the reducing reagents. The reaction proceeds automatically without any enzymes or reagents under biological conditions to produce a fluorescence signal within 10-20 min in the presence of target DNA or RNA. In addition, the probe was very stable under biological conditions, even such extreme conditions as pH 5 solution, pH 10 solution, or high temperature (90°C) with no undesirable background signal. The probes were successfully applied to the detection of oligonucleotides at the single nucleotide level in solution and endogenous RNA in bacterial cells. © 2008 American Chemical Society.

DOI： 10.1021/bc800014d

MEDI 134-Multiple chemical ligation under thermal cycle

Kondo Yuko, Abe Hiroshi, Jinmei Hiroshi, Abe Naoko, Aikawa Kyoko, Matsumoto Isamu, Ito Yoshihiro

ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 235 巻 2008年4月

Rhodamine-based fluorogenic probe for imaging biological thiol 査読有り

Shibata Aya, Furukawa Kazuhiro, Abe Hiroshi, Tsuneda Satoshi, Ito Yoshihiro

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 18 巻 ( 7 ) 頁： 2246 - 2249 2008年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioorganic and Medicinal Chemistry Letters

We have developed a new fluorescent probe for biological thiol. The probe was synthesized by the modification of the 2,4-dinitrobenzenesulfonyl group with rhodamine 110. The selective detection of thiol species such as cysteine or glutathione was achieved in biological conditions. Moreover, the probe was successfully applied to the imaging of thiol species in living human cells. © 2008 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmcl.2008.03.014

Synthesis of (+/-)-9-[c-4, t-5-bis(hydroxymethyl)cyclopent-2-en-r-1-yl]-9H-adenine (BCA) derivatives branched at the 4 '-position based on intramolecular SH2 ' cyclization 査読有り

Kumamoto Hiroki, Takahashi Nonoko, Shimamura Tomomi, Tanaka Hiromichi, Nakamura Kazuo T., Hamasaki Takayuki, Baba Masanori, Abe Hiroshi, Yano Masahiko, Kato Nobuyuki

TETRAHEDRON 64 巻 ( 7 ) 頁： 1494 - 1505 2008年2月

Chemical aminoacylation of RNA by an intermolecular adenosine transfer reaction 査読有り

Liu Mingzhe, Jinmei Hiroshi, Abe Hiroshi, Ito Yoshihiro

CHEMISTRY LETTERS 37 巻 ( 1 ) 頁： 102 - 103 2008年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Chemistry Letters

RNA aminoacylation, based on an adenosine transfer mechanism, was achieved by the intermolecular transfer of phenylalaninyl adenosine from a donor probe to an acceptor RNA. This method can be applied to a wide variety of unnatural amino acids for tRNA aminoacylation. Copyright © 2008 The Chemical Society of Japan.

DOI： 10.1246/cl.2008.102

Stereoselective glycosylation based on the conformational restriction of pyranoses 査読有り

Satoshi Shuto, Satoshi Ichikawa, Hiroshi Abe, Akira Matsuda

JOURNAL OF SYNTHETIC ORGANIC CHEMISTRY JAPAN 66 巻 ( 1 ) 頁： 50 - 60 2008年1月

記述言語：日本語出版者・発行元：SOC SYNTHETIC ORGANIC CHEM JPN

Despite considerable progress and extensive effort, a general method for highly stereoselective glycosylation particularly for the 1,2-cis-glycosylation has not yet been developed and therefore is required. The alpha/beta-stereo selectivity in glycosylation can be affected by the steric and stereoelectronic (anomeric) effects around the anomeric center, which depend on the conformation of the glycosyl donor substrates. Therefore, we hypothesized that highly alpha- and beta-selective glycosylation can be realized by employing conformationally restricted substrates. We showed that the alpha/beta-stereoselectivity was significantly increased by the conformational restriction and was completely inverted by changing the substrate conformation from the C-4(1)-form into the C-1(4)-form in radical and nucleophilic C-glycosylation reactions as well as in O-glycosylation reactions. The conformational restriction of substrates also effectively facilitates the alpha- and beta-selective radical cyclization reaction at the anomeric position. Using the method, C-glucoside trisphosphates designed as Ca2+-mobilizing agents were successfully,synthesized.

DOI： 10.5059/yukigoseikyokaishi.66.50

Rapid DNA chemical ligation for amplification of RNA and DNA signal 査読有り

Abe Hiroshi, Kondo Yuko, Jinmei Hiroshi, Abe Naoko, Furukawa Kazuhiro, Uchiyama Atsushi, Tsuneda Satoshi, Aikawa Kyoko, Matsumoto Isamu, Ito Yoshihiro

BIOCONJUGATE CHEMISTRY 19 巻 ( 1 ) 頁： 327 - 333 2008年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioconjugate Chemistry

Enzymatic ligation methods are useful in the diagnostic detection of DNA sequences. Here, we describe the investigation of nonenzymatic phosphorothioate-iodoacetyl DNA chemical ligation as a method for the detection and identification of RNA and DNA. The specificity of ligation on the DNA target is shown to allow the discrimination of a single point mutation with a drop in the ligation yield of up to 16.1-fold. Although enzymatic ligation has very low activity for RNA targets, this reaction is very efficient for RNA targets. The speed of the chemical ligation with an RNA target achieves a 70% yield in 5 s, which is equal to or better than that of ligase-enzyme-mediated ligation with a DNA target. The reaction also exhibits a significant level of signal amplification under thermal cycling in periods as short as 100-120 min, with the RNA or DNA target acting in a catalytic way to ligate multiple pairs of probes. © 2008 American Chemical Society.

DOI： 10.1021/bc700244s

In vitro selection of RNA aptamer to hemin. 査読有り

Mingze Liu, Takuma Kagahara, Hiroshi Abe, Yoshihiro Ito

Nucleic Acids Symposium Series 52 巻頁： 513 - 514 2008年

Fluorogenic probe triggered by reduction for nucleic acids sensing. 査読有り

Furukawa K, Abe H, Wang J, Oki K, Uda M, Tsuneda S, Ito Y

Nucleic acids symposium series (2004) ( 52 ) 頁： 353-4 2008年

Quenched Autoligation Probes 査読有り

Adam P. Silverman, Hiroshi Abe, Eric T. Kool

Methods in Molecular Biology 429 巻頁： 161 - 170 2008年

細胞内遺伝子シグナルの解析

阿部洋、古川和寛、常田聡、伊藤嘉浩

生物工学、日本生物工学会 86 巻 ( 6 ) 頁： 268-270 2008年

Quenched autoligation probes

Adam P. Silverman, Hiroshi Abe, Eric T. Kool

Methods in Molecular Biology 頁： 161-170 2008年

ピラノースの配座制御に基づく立体選択的グリコシル化反応

周東智、市川聡、阿部洋、松田彰

有機合成化学協会誌頁： 50-60 2008年

ダンベル型ナノサークルRNAでRNA干渉を長期安定に

阿部洋、阿部奈保子、原田充、常田聡、伊藤嘉浩

バイオインダストリー協会 66 巻 ( 8 ) 頁：全 2008年

Design and synthesis of biorelated polymers by combinatorial bioengineering

Yoshihiro Ito, Hiroshi Abe, Akira Wada, Mingahe Liu

ACS Symposium Series999(Polymer Biocatalysis and Biomaterials 2 ) 頁： 194-215 2008年

Nanocircular RNAs for RNA interference 査読有り

Hiroshi Abe, Naoko Abe, Mitsuru Harada, Satoshi Tsuneda, Yoshihiro Ito

Nucleic Acids Symposium Series 52 巻頁： 505 - 506 2008年

Fluorogenic probe triggered by reduction for nucleic acids sensing 査読有り

Kazuhiro Furukawa, Hiroshi Abe, Jin Wang, Kazuma Oki, Miwako Uda, Satoshi Tsuneda, Yoshihiro Ito

Nucleic Acids Symposium Series 52 巻頁： 353 - 354 2008年

Design and Synthesis of Biorelated Polymers by Combinatorial Bioengineering

Ito Yoshihiro, Abe Hiroshi, Wada Akira, Liu Mingzhe

POLYMER BIOCATALYSIS AND BIOMATERIALS II 999 巻頁： 194 - 215 2008年

In vitro selection of RNA aptamer to hemin. 査読有り

Liu M, Kagahara T, Abe H, Ito Y

Nucleic acids symposium series (2004) ( 52 ) 頁： 513 - 4 2008年

Nanocircular RNAs for RNA interference. 査読有り

Abe H, Abe N, Harada M, Tsuneda S, Ito Y

Nucleic acids symposium series (2004) ( 52 ) 頁： 505 - 6 2008年

Dumbbell-shaped nanocircular RNAs for RNA interference 査読有り

Abe Naoko, Abe Hiroshi, Ito Yoshihiro

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 129 巻 ( 49 ) 頁： 15108 - + 2007年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of the American Chemical Society

We designed and synthesized dumbbell-shaped nanocircular RNAs for RNA interference applications, which consist of a stem and two loops. RNA dumbbells are specifically recognized and cleaved by the human Dicer enzyme and are thus transformed into double-stranded RNA in cells, although this RNA is resistant to degradation in serum. The structure was optimized to maximize its RNAi activity. The most potent activity was achieved when the stem length was 23 base pairs. The RNAi activity is prolonged by the shape of the molecule, an endless structure, compared with that of normal siRNA. Copyright © 2007 American Chemical Society.

DOI： 10.1021/ja0754453

[Bioprobe for imaging RNA in living cells]. 査読有り

Abe H, Furukawa K, Tsuneda S, Ito Y

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 52 巻 ( 13 Suppl ) 頁： 1619-24 2007年10月

Intermolecular transfer of aminoacylated adenosine towards chemical aminoacylation of tRNA 査読有り

Mingze Liu, Hiroshi Jinmei, Koji Fukuzono, Hiroshi Abe, Yoshihiro Ito

Nucleic Acids Symposium Series 51 巻頁： 353 - 354 2007年

Multiple chemical ligation under thermal cycle. 査読有り

Yuko Kondo, Hiroshi Abe, Hiroshi Jinmei, Naoko Abe, Kyoko Aikawa, Isamu Matsumoto, Yoshihiro Ito

Nucleic Acids Symposium Series 51 巻頁： 365 - 366 2007年

Multiple chemical ligation under thermal cycle. 査読有り

Kondo Y, Abe H, Jinmei H, Abe N, Aikawa K, Matsumoto I, Ito Y

Nucleic acids symposium series (2004) ( 51 ) 頁： 353 - 4 2007年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic acids symposium series (2004)

Enzymatic ligation methods are useful in diagnostic detection of DNA sequence. Here we describe the investigation of nonezymatic phosphorothioate-iodoacetyl DNA chemical ligation as a method for detection and identification of RNA and DNA. Specificity of ligation on DNA target is shown to yield discrimination of single point mutation as a drop in two magnitude. Although enzymatic ligation shows very low activity for RNA target, this reaction is found to be very efficient on RNA target. This chemical ligation with RNA target completes 70% within a few seconds, which equal or overcome ligase enzyme-mediated ligation with DNA target. The reaction is also shown to exhibit a significant level of signal amplification under thermal cycle for short time. Further, we found recently that ligation fidelity changed in function of chemical reactivity of probe. This trend was systematically investigated.

DOI： 10.1093/nass/nrm177

Intermolecular transfer of aminoacylated adenosine towards chemical aminoacylation of tRNA. 査読有り

Liu M, Jinmei H, Fukuzono K, Abe H, Ito Y

Nucleic acids symposium series (2004) ( 51 ) 頁： 365 - 6 2007年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic acids symposium series (2004)

We synthesized two oligonucleotides: one is DNA conjugated with adenosine derivative at 5' terminus as a donor, another RNA phosphorothioated at 3' terminus as an acceptor. By hybridization of the two oligonuleotides, the adenosine derivative was transferred from the donor DNA to the acceptor RNA. In this study, O, O'-isopropylidene-protected adenosine derivative and aminoacylated adenosine were successfully transferred from the donor to acceptor RNA. By use of tRNA instead of the acceptor RNA, the chemical aminoacylation of tRNA by unnatural amino acid is expected.

DOI： 10.1093/nass/nrm183

Synthesis of adenophostin A analogues conjugating an aromatic group at the 5 '-position as potent IP3 receptor ligands 査読有り

Tetsuya Mochizuki, Yoshihiko Kondo, Hiroshi Abe, Stephen C. Tovey, Skarlatos G. Dedos, Colin W. Taylor, Michael Paul, Barry V. L. Potter, Akira Matsuda, Satoshi Shuto

JOURNAL OF MEDICINAL CHEMISTRY 49 巻 ( 19 ) 頁： 5750 - 5758 2006年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

Previous structure-activity relationship studies of adenophostin A, a potent IP3 receptor agonist, led us to design the novel adenophostin A analogues 5a-c, conjugating an aromatic group at the 5'-position to develop useful IP3 receptor ligands. The common key intermediate, a D-ribosyl alpha-D-glucoside 10 alpha, was stereoselectively synthesized by a glycosidation with the 1-sulfinylglucoside donor 11, which was conformationally restricted by a 3,4-O-cyclic diketal protecting group. After introduction of an aromatic group at the 5-position of the ribose moiety, an adenine base was stereoselectively introduced at the anomeric beta-position to form 7a-c, where the tetra-O-i-butyryl donors 9a-c were significantly more effective than the corresponding O-acetyl donor. Thus, the target compounds 5a-c were synthesized via phosphorylation of the 2', 3", and 4"-hydroxyls. The potencies of compounds 5a-c for Ca2+ release were shown to be indistinguishable from that of adenophostin A, indicating that bulky substitutions at the 5'-position of adenophostin A are well-tolerated in the receptor binding. This biological activity of 5a-c can be rationalized by molecular modeling using the ligand binding domain of the IP3 receptor.

DOI： 10.1021/jm060310d

Design and synthesis of 5'-deoxy-5'-phenyladenophostin A, a highly potent IP3 receptor ligand. 査読有り

Mochizuki T, Kondo Y, Abe H, Taylor CW, Potter BV, Matsuda A, Shuto S

Organic letters 8 巻 ( 7 ) 頁： 1455-8 2006年3月

Design and synthesis of 5 '-deoxy-5 '-phenyladenophostin A, a highly potent IP3 receptor ligand 査読有り

T Mochizuki, Y Kondo, H Abe, CW Taylor, BVL Potter, A Matsuda, S Shuto

ORGANIC LETTERS 8 巻 ( 7 ) 頁： 1455 - 1458 2006年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

5'-Deoxy-5'-phenyladenophostin A (5), designed as a useful IP3 receptor ligand based on the previous structure-activity relationship studies, was successfully synthesized via two key stereoselective glycosidation steps. This compound proved to be a highly potent IP3 receptor agonist.

DOI： 10.1021/o10602710

A systematic study of C-glucoside trisphosphates as myo-inositol trisphosphate receptor ligands. Synthesis of beta-C-glucoside trisphosphates based on the conformational restriction strategy 査読有り

M Terauchi, H Abe, SC Tovey, SG Dedos, CW Taylor, M Paul, M Trusselle, BVL Potter, A Matsuda, S Shuto

JOURNAL OF MEDICINAL CHEMISTRY 49 巻 ( 6 ) 頁： 1900 - 1909 2006年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

beta-C-Glucoside trisphosphates having a C2 side chain (3,7-anhydro-2-deoxy-D-glycero-D-gulo-octitol 1,5,6-trisphosphate, 11) and a C3 side chain (4,8-anhydro-2,3-dideoxy-D-glycero-D-gulo-nonanitol 1,6,7-trisphosphate, 12) were designed as structurally simplified analogues of a potent D-myo-inositol 1,4,5-trisphosphate (IP3) receptor ligand, adenophostin A. Construction of the beta-C-glucosidic structure, which was the key to their synthesis, was achieved by two different methods based on the conformational restriction strategy: (1) radical cyclization with a temporary connecting silicon tether and (2) silane reduction of glyconolactols having an anomeric allyl substituent. Using these methods, the target beta-C-glycoside trisphosphates 11 and 12 were successfully synthesized. A structure-activity relationship was established on a series of C-glucoside trisphosphates, including the previously synthesized related compounds, which were a C-glycosidic analogue 3 of adenophostin A, its uracil congener 5, alpha-C-glucoside trisphosphates 7-9 having a C1, C2, or C3 side chain, and the beta-C-glucoside trisphosphates 10-12 having a C1, C2, or C3 side chain. The O-glycosidic linkage of adenophostin A and its analogues proved to be replaced by the chemically and biologically more stable C-glycosidic linkage. The alpha-C2-glucoside trisphosphate 8 stimulates Call release with a potency similar to that of IP3 in spite of its simplified structure, indicating a better fit to the receptor than the beta-C-glucoside trisphosphates and also the alpha-congeners having a shorter or longer C1 side chain, which was supported by molecular modeling using the ligand binding domain of the IP3 receptor.

DOI： 10.1021/jm051039n

Flow cytometric detection of specific RNAs in native human cells with quenched autoligating FRET probes. 査読有り

Abe H, Kool ET

Proceedings of the National Academy of Sciences of the United States of America 103 巻 ( 2 ) 頁： 263-8 2006年1月

Structure analysis of oligonucleotide in organic solvent 査読有り

Hiroshi Abe, Naoko Abe, Yoshihiro Ito

Nucleic Acids Symposium Series 50 巻頁： 25 - 26 2006年

Structure analysis of oligonucleotide in organic solvent. 査読有り

Abe H, Abe N, Ito Y

Nucleic acids symposium series (2004) ( 50 ) 頁： 25 - 6 2006年

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nucleic acids symposium series (2004)

Oligonucleotide has been used as functional molecule such as recording device, or catalyst for chemical reaction. However, that utility has been limited, because it is only soluble in water. Recently, we succeeded in making chemically modified DNA which is clearly soluble in organic solvent. Using this technique, G-quadruplex was constructed in non-polar organic solvent such as 1,2-dichloroethane. The structure was analyzed by CD spectra.

DOI： 10.1093/nass/nrl013

新機能性核酸・タンパク質を生み出すコンビナトリアル・バイオエンジニアリング

阿部洋、和田章、伊藤嘉浩

生物工学会誌 84 巻 ( 8 ) 頁： 309 2006年

Synthesis of 4,8-anhydro-D-glycero-D-ido-nonanitol 1,6,7-trisphosphate as a novel IP3 receptor ligand using a stereoselective radical cyclization reaction based on a conformational restriction strategy 査読有り

M Terauchi, Y Yahiro, H Abe, S Ichikawa, SC Tovey, SG Dedos, CW Taylor, BVL Potter, A Matsuda, S Shuto

TETRAHEDRON 61 巻 ( 15 ) 頁： 3697 - 3707 2005年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

4,8-Anhydro-D-glycero-D-ido-nonanitol 1,6,7-trisphosphate (9), designed as a novel IP3 receptor ligand having an alpha-C-glycosidic\ structure, was synthesized via a radical cyclization reaction with a temporary connecting allylsilyl group as the key-step. Phenyl 2-O-allyldimethylsilyt-3,4-bis-O-TBS-1-seleno- beta-D-glucopyranoside (10a), conformationally restricted in the unusual C-1(4)-conformation, was treated with Bu3SnH/AIBN to form the desired alpha-cyclization product 16a almost quantitatively. On the other hand, when a conformationally unrestricted O-benzyl-protected 2-O-allyldimethylsilyl -l-selenoglucoside 15 was used as the substrate, the radical reaction was not stereoselective and gave a mixture of the alpha-and beta-products. From 16a, the target C-glucoside trisphosphate 9 was synthesized via phosphorylation of the hydroxyls by the phosphoramidite method. During the synthetic study, an efficient procedure for the oxidative C-Si bond cleavage, via a nucleophilic substitution at the silicon with p-MeOPhLi followed by Fleming oxidation, was developed. The C-glycoside 9 was found to be a full agonist for Ca2+ mobilization, although its activity was weaker than that of the natural ligand IP3. Thus, the alpha-C-glucosidic structure was shown to be a useful mimic of the myo-inositol backbone of IP3. (c) 2005 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tet.2005.02.025

Universal linkers for signal amplification in auto-ligating probes 査読有り

Hiroshi Abe, Eric T. Kool

Nucleic Acids Symposium Series 49 巻頁： 37 - 38 2005年

Universal linkers for signal amplification in auto-ligating probes. 査読有り

Abe H, Kool ET

Nucleic acids symposium series (2004) ( 49 ) 頁： 37-8 2005年

Highly stereoselective Grignard addition to cis-substituted C-cyclopropylaldonitrones. The bisected s-trans transition state can be stabilized effectively by the Lewis acid-coordination 査読有り

Y Kazuta, H Abe, A Matsuda, S Shuto

JOURNAL OF ORGANIC CHEMISTRY 69 巻 ( 26 ) 頁： 9143 - 9150 2004年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

We previously found that Grignard addition to a C-cyclopropylaldonitrone, C-[cis-2-(N,N-diethyl-carbamoyl)-trans-2-phenylcyclopropyl]-N-benzylaldonitrone (1), stereoselectively gave the antiproduct 3, in which the stereoselectivity was particularly high when MgBr2 was the additive. In this study, the reaction pathway was investigated in detail. The stereoselective addition was initially thought to occur via either a 1,5-chelation-controlled or a bisected s-trans conformation-controlled pathway. However, Grignard addition to a nonchelating silyl ether-type substrate, C-[cis-2-(tert-butyldiphenylsilyloxymethyl)-trans-2-phenylcyclopropyl]-N-benzylaldonitrone (7), also gave the antiproduct 9 with high stereoselectivity suggesting that chelation is not important in the reaction. Theoretical calculations of C-cyclopropylaldonitrones showed that the coordination of Mg2+ at the nitrone oxygen significantly stabilizes the bisected s-trans conformer due to the effective hyper-conjugation between the,pi* of the nitrone C=N bond and the electron-donating cyclopropane orbitals. This kind of orbital interaction is able to stabilize the transition state of the nucleophilic addition and is maximized in the bisected conformation, in which the orbitals of the forming bond and the cyclopropane C-C bond are in an almost planar arrangement. Thus, the high stereoselectivity can be explained by nucleophilic attack on the less hindered side of the C=N bond of the substrates in the Mg2+-coordinated bisected s-trans conformation.

DOI： 10.1021/jo048637e

Destabilizing universal linkers for signal amplification in self-ligating probes for RNA. 査読有り

Abe H, Kool ET

Journal of the American Chemical Society 126 巻 ( 43 ) 頁： 13980-6 2004年11月

An efficient synthesis of beta-C-glycosides based on the conformational restriction strategy: Lewis acid promoted silane reduction of the anomeric position with complete stereoselectivity 査読有り

M Terauchi, H Abe, A Matsuda, S Shuto

ORGANIC LETTERS 6 巻 ( 21 ) 頁： 3751 - 3754 2004年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

The reduction of glyconolactols having an anomeric carbon substituent by Et3SiH/TMSOTf proceeded with complete stereoselectivity to produce the corresponding beta-C-glycosides when the substrates were conformationally restricted in the C-4(1)-chair form by a 3,4-O-cyclic diketal or a 4,6-O-benzylidene protecting group. Thus, the efficient construction beta-C-glycosides was achieved on the basis of the conformation restriction strategy.

DOI： 10.1021/ol048525+

Stereoselective synthesis of α- and β-C-glucosides via radical cyclization with an allylsilyl tether. Control of the stereoselectivity by changing the conformation of the pyranose ring. 査読有り

Satoshi Shuto, Masaru Terauchi, YumiYahiro, Hiroshi Abe, Satoshi Ichikawa, Akira Matsuda

Tetrahedron Letters 45 巻 ( 36 ) 頁： 6819 - 6819 2004年8月

The unusual 1,4-chelation-controlled nucleophilic addition to aldehydes with high stereoselectivity. A systematic study of stereoselectivity in the addition reaction of carbon nucleophiles to cis-substituted cyclopropanecarbaldehydes 査読有り

Y Kazuta, H Abe, T Yamamoto, A Matsuda, S Shuto

TETRAHEDRON 60 巻 ( 31 ) 頁： 6689 - 6703 2004年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

The addition reaction of carbon nucleophiles to cis-substituted cyclopropanecarbaldehydes was systematically investigated. Ab initio calculations of model cyclopropanecarbaldehydes suggested that the bisected s-cis and s-trans conformers are the only two minimum energy conformers, which are stabilized due to the pi-donating stereoelectronic effect of the cyclopropane ring. The experimental results of a series of substrates, that is, cyclopropanecarbaldehydes 1-5 bearing a cis-(tert-butyldiphenyisilyloxy)methyl group, a cis-benzyloxymethyl group, a cis-(p-methoxybenzyloxy)methyl group, cis-N,N-diethylcarbamoyl and trans-phenyl groups, and cis-(tert-butyldiphenylsilyloxy)methyl and trans-phenyl groups, respectively, showed that highly anti-selective Grignard additions could be realized. It turned out that it occurred via an unusual 7-membered 1,4-chelation-controlled pathway. Highly stereoselective Grignard addition via the chelation-controlled pathway occurred even in the reaction of the usually non-chelating silyl ether-type substrate 5. The results have great importance because the 1,4-chelation-controlled stereoselective addition reactions can indeed be realized. Under non-chelation conditions, the syn-products were produced with moderate stereoselectivity, which are likely to be formed via the bisected s-cis conformation-like transition state stabilized by the characteristic orbital interaction. These reactions, especially the chelation-controlled reaction, should be useful because of their t stereoselectivity and stereochemical predictability. (C) 2004 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tet.2004.05.063

Quenched auto-ligating DNAs: Multicolor identification of nucleic acids at single nucleotide resolution 査読有り

S Sando, H Abe, ET Kool

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 126 巻 ( 4 ) 頁： 1081 - 1087 2004年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

We describe the synthesis and study of multicolor quenched autoligating (QUAL) probes for identification and discrimination of closely related RNA and DNA sequences in solution and in bacteria. In these probes, a dabsyl quencher doubles as an activator in the oligonucleotide-joining reaction. The oligonucleotides remain dark until they bind at adjacent sites, and "light up" on nucleophilic displacement of the dabsyl probe by the phosphorothioate probe. Four fluorescent dye conjugates were prepared and tested with probes and targets that differ by one nucleotide. Experiments on polymer beads show clear color-based discrimination of DNAs added in solution. Two-color quenched probe pairs were then tested in the discrimination of 16S rRNA sequences in Escherichia coli. Single nucleotide resolution was achieved in the cells with green/red QUAL probes, allowing identification of a one-base sequencing error in the 16S rRNA database. Finally, QUAL probes were successfully applied in live bacterial cells. The method requires only incubation followed by fluorescence imaging, and requires no enzymes, added reagents, cross-linking, fixing, or washes. Because probes must bind side-by-side to generate signal, there is little or no interference from unintended protein binding, which can occur with other probe types. The results suggest that QUAL probes may be of general use in the detection and identification of sequences in solution, on microarrays, and in microorganisms.

DOI： 10.1021/ja038665z

Construction of a cis-cyclopropane via reductive radical decarboxylation. Enantioselective synthesis of cis- and trans-1-arylpiperazyl-2-phenyleyclopropanes designed as antidopaminergic agents 査読有り

K Yamaguchi, Y Kazuta, H Abe, A Matsuda, S Shuto

JOURNAL OF ORGANIC CHEMISTRY 68 巻 ( 24 ) 頁： 9255 - 9262 2003年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

(1S,2S)-, (1S,2R)-, and (1R,2S)-1-(2,4-Dimethylphenyl)piperazyl-2-phenylcyclopropane (2a, 3, and ent-3, respectively), which were designed as conformationally restricted analogues of haloperidol (1), a clinically effective antipsychotic agent, were synthesized from chiral epichlorohydrins using the Barton reductive radical decarboxylation as the key step. (1S,2R)-1-(tert-Butyldiphenylsilyloxy)methyl-2-carboxy-2-phenylcyclopropane (5), which was prepared from (S)-epichlorohydrin ((S)-7), was converted into its N-hydroxypyridine-2-thione ester 12, the substrate for the reductive radical decarboxylation. When 12 was treated with TMS(3)SiH in the presence of Et(3)B or AIBN, the decarboxylation and subsequent hydride attack on the cyclopropyl radical intermediate from the side opposite to the bulky silyloxymethyl moiety occurred, resulting in selective formation of the corresponding reductive decarboxylation product 4-cis with the cis-cyclopropane structure. From 4-cis, the cis-cyclopropane-type target compound 3 was readily synthesized. Starting from (R)epichlorohydrin ((R)-7), ent-3 was similarly synthesized. Epimerization of the cyclopropanecarboxamide ent-16-cis, a synthetic intermediate for ent-3, on treatment with a base prepared from Bu(2)Mg and i-Pr(2)NH in THF occurred effectively to give the corresponding trans isomer 16-trans, which was converted into 2a with the trans-cyclopropane structure.

DOI： 10.1021/jo0302206

A study on the conformation-anomeric effect-stereoselectivity relationship in anomeric radical reactions, using conformationally restricted glucose derivatives as substrates 査読有り

H Abe, M Terauchi, A Matsuda, S Shuto

JOURNAL OF ORGANIC CHEMISTRY 68 巻 ( 19 ) 頁： 7439 - 7447 2003年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

We previously theorized that, since the stereoselectivity of anomeric radical reactions is significantly influenced by the kinetic anomeric effect, which can be controlled by restricting the conformation of the radical intermediate, the proper conformational restriction of the pyranose ring of the substrates would therefore make highly alpha- and beta-stereoselective anomeric radical reactions possible. This theory was based on our previous results of the anomeric radical reactions with D-xylose derivatives as the substrates. We herein report the anomeric radical deuteration reactions with the conformationally restricted 1-phenylseleno-D-glucose derivatives, 2g and 3g, restricted in a C-4(1)-conformation by an beta-cyclic diketal moiety, and 4g, 5g, 6g, 7g, and 8g, restricted in a C-1(4)-conformation by bulky O-silyl protecting groups. The radical deuterations with Bu3SnD, using the C-4(1)-restricted substrates 2g and 3g, afforded the corresponding alpha-products (alpha/beta = 98:2) highly stereoselectively, whereas the C-1(4)-restricted substrate 6g, having a trigonal (sp(2)) carbon substituent, i.e., -CHO, at the 5-position, selectively gave the beta-products (alpha/beta = 0: 100). Thus, the stereoselectivity was significantly increased by the conformational restriction and was completely inverted by changing the substrate conformation from the C-4(1)-form to the C-1(4)-form. On the other hand, the deuterations with the C-1(4)-restricted substrates 4g and 5g showed that the 1,5-steric effect due to the tetrahedral carbon substituent (-CH2OTIPS or -CH2OH) at the 5-axial position dominantly prevented the hydride transfer from the beta-face competing with the kinetic anomeric effect. This study suggests that, depending on the restricted conformation of the substrates to the C-4(1)- or the C-1(4)-form, the alpha- or beta-products would be obtained highly stereoselectively via anomeric radical reactions of hexopyranoses.

DOI： 10.1021/jo030128+

A systematic study of the hydride reduction of cyclopropyl ketones with structurally simplified substrates. Highly stereoselective reductions of trans-substituted cyclopropyl ketones via the bisected s-cis conformation 査読有り

Y Kazuta, H Abe, T Yamamoto, A Matsuda, S Shuto

JOURNAL OF ORGANIC CHEMISTRY 68 巻 ( 9 ) 頁： 3511 - 3521 2003年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

The stereoselective hydride reduction of the cis- and trans-substituted cyclopropyl ketones was systematically investigated using a series of structurally simplified substrates, trans-[tert-butyldiphenylsilyloxymethyl]cyclopropyl ketones la-a and trans- (benzyloxymethyl)cyclopropyl methyl ketone (2), and the corresponding cis congeners 3a,b,e and 4. The results showed that, not only in the reduction of the cis-substituted cyclopropyl ketones but also in that of the trans-substituted ketones, high stereoselectivity can be realized when the substrate has a bulky substituent on the cyclopropane ring, even though it is attached to the position trans to the acyl moiety. Ab initio calculations based on the density functional theory (DFT) of cyclopropyl ketones showed that (1) the bisected s-cis and s-trans conformers were the only two minimum energy conformers; while the s-cis conformer was more stable than the s-trans and (2) a bulky alkyl group in the acyl moiety and a cis substituent on the cyclopropane ring made the bisected s-cis conformer much more stable. On the basis of these calculations and experimental results, it is likely that the more stable the bisected s-cis conformer of the substrate, the more stereoselective the hydride reduction. Thus, the stereochemistry can be explained by hydride attack on the bisected s-cis conformation of the substrate from the less-hindered face. The predictability of the stereochemical results is predicated on the bisected s-cis transition-state model, which is very important from the viewpoint of synthetic organic chemistry.

DOI： 10.1021/jo030019v

Control of alpha/beta stereoselectivity in Lewis acid promoted C-glycosidations using a controlling anomeric effect based on the conformational restriction strategy 査読有り

S Tamura, H Abe, A Matsuda, S Shuto

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 42 巻 ( 9 ) 頁： 1021 - + 2003年

Development of New Radical Reactions with a Vinylsilyl Group and Their Application to the Synthesis of Branched-Chain Sugar Nucleosides

Satoshi Shuto, Makiko Kanazaki, Isamu Sugimoto, Satoshi Ichikawa, Yuki Nagasawa, Yoshihito Ueno, Hiroshi Abe, Noriaki Minakawa, Makoto Sukeda, Tetsuya Kodama, Makoto Nomura, Akira Matsuda

Recent Advances in Nucleosides: Chemistry and Chemotherapy 頁： 21 - 55 2002年

Synthesis and biological activity of adenophostin derivatives as IP3 receptor ligands. 査読有り

Shuto S, Mochizuki T, Abe H, Kondo Y, Matsuda A

Nucleic acids research. Supplement (2001) ( 2 ) 頁： 23-4 2002年

Development of new radical reactions with a vinylsilyl group and their application to the synthesis of branched-chain sugar nucleosides

Satoshi Shuto, Maki kanazaki, Isamu Sugimoto, Satoshi Ichikawa, Yuki Nagasawa, Yoshihito Ueno, Hiroshi Abe, Noriaki Minakawa, Makoto Sukeda, Tetsuya Kodama, Makoto Nomura, Akira matsuda

Recent Advances in Nucleosides 頁： 21-55 2002年

Synthesis and biological activity of adenophostin derivatives as IP3 receptor ligands 査読有り

Satoshi Shuto, Tetsuya Mochizuki, Hiroshi Abe, Yoshihiko Kondo, Akira Matsuda

Nucleic Acids Symposium Series 2 巻頁： 23 - 24 2002年

Highly alpha- and beta-selective radical C-glycosylation reactions using a controlling anomeric effect based on the conformational restriction strategy. A study on the conformation-anomeric effect-stereoselectivity relationship in anomeric radical reactions 査読有り

H Abe, S Shuto, A Matsuda

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 123 巻 ( 48 ) 頁： 11870 - 11882 2001年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

We hypothesized that, because the stereoselectivity of anomeric radical reactions was significantly influenced by the anomeric effect, which can be controlled by restricting the conformation of the radical intermediate, the proper conformational restriction of the pyranose ring of the substrates would therefore make highly alpha- and beta -stereoselective anomeric radical reactions possible. Thus, the conformationally restricted 1-phenylseleno-D-xylose derivatives 9 and 10, restricted in a C-4(1)-conformation, and 11 and 12, restricted in a C-1(4)-conformation, were designed and synthesized by introducing the proper protecting groups on the hydroxyl groups on the pyranose ring as model substrates for the anomeric radical reactions. The radical deuterations with Bu3SnD and the C-glycosylation with Bu3SnCH2CH=CH2 or CH2 CHCN, using the C-4(1)-restricted substrates 9 and 10, afforded the corresponding alpha -products (alpha/beta = 97:3-85:15) highly stereoselectively, whereas the C-1(4)-restricted substrates 11 and 12 selectively gave the fl-products (alpha/beta = 1:99-0:100). Thus, stereoselectivity was significantly increased by conformational restriction and was completely inverted by changing the substrate conformation from the C-4(1)-form into the C-1(4)-form. Ab initio, calculations suggested that the radical intermediates produced from these substrates possessed the typical C-4(1)- or C-1(4)-Conformation, which was similar to that of the substrates, and that the anomeric effect in these conformations would be the factor controlling the transition state of the reaction. Therefore, the highly alpha- and beta -selective reactions would occur because of the anomeric effect, which could be manipulated by conformational restriction of the substrates, as expected. This would be the first radical C-glycosylation reaction to provide both alpha- and beta -C-glycosides highly stereoselectively.

DOI： 10.1021/ja011321t

An efficient method for preparing fully O-silylated pyranoses conformationally restricted in the unusual C-1(4)-form 査読有り

H Abe, S Shuto, S Tamura, A Matsuda

TETRAHEDRON LETTERS 42 巻 ( 35 ) 頁： 6159 - 6161 2001年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

An efficient method for preparing fully O-silylated pyranoses, which are conformationally restricted in the unusual C-1(4)-form, was developed. Thus, successive treatment of pyranosides, such as xylose and glucose derivatives, with NaH and TIPSOTf or TBSOTf in THF at room temperature gave the corresponding fully O-silylated products. (C) 2001 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0040-4039(01)01233-3

Significance of substrate hydrophobicity for recognition by an oligopeptide transporter (PEPT1) 査読有り

R Tateoka, H Abe, S Miyauchi, S Shuto, A Matsuda, M Kobayashi, K Miyazaki, N Kamo

BIOCONJUGATE CHEMISTRY 12 巻 ( 4 ) 頁： 485 - 492 2001年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

Our previous paper [(1999) Bioconjugate Chem. 10, 24-31] pointed out that hydrophobicity of substrates/inhibitors plays an important role in the recognition by an oligopeptide transporter (PEPT1) expressed in the human intestinal epithelial cell line Caco-8. To determine the significance of that hydrophobicity, we have now synthesized dipeptide analogues conjugating the E-amino group of Lys in Val-Lys with aliphatic carboxylic acids: acetic acid (C2), propanoic acid (C3), pentanoic acid (C5), hexanoic acid (C6), and decanoic acid (C10). The affinities of these conjugates were estimated by their inhibition of the accumulation rate of Gly-Sar, a well-established substrate for PEPT1. With the increase in length of the hydrocarbon chain of the conjugates, i.e., in the hydrophobicity of the conjugates, the inhibition strengthened. Dixon-Webb plot analysis of the inhibition by the C10-conjugated dipeptide showed competitive inhibition. The trans-stimulation effect of Val-Lys conjugated to C10 or C5 on the uptake of Ceftibuten was observed using rat brush border membrane vesicles. This findings showed that these conjugates are transportable substrates. These results confirmed that the hydrophobicity of substrates/inhibitor is one of the factors in the recognition by PEPT1.

DOI： 10.1021/bc000135u

INTRACELLULAR Ca2+-MOBILIZING ADENINE NUCLEOTIDES. SYNTHESIS AND BIOLOGICAL ACTIVITY OF CYCLIC ADP-CARBOCYCLIC-RIBOSE ANDC-GLYCOSIDIC ANALOG OF ADENOPHOSTIN A

Satoshi Shuto, Masayoshi Fukuoka, Hiroshi Abe, Akira Matsuda

Nucleosides, Nucleotides and Nucleic Acids 20 巻 ( 4-7 ) 頁： 461 - 470 2001年3月

The bisected s-trans conformation-controlled highly stereoselective addition of Grignard reagents to C-cyclopropylaldonitrone. An efficient synthesis of 1-phenyl-2-[(S)-1-aminoalkyl]-N,N-diethylcyclopropanecarboxamides, a new class of potent NMDA receptor antagonists 査読有り

Y Kazuta, S Shuto, H Abe, A Matsuda

JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1 1 巻 ( 6 ) 頁： 599 - 604 2001年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ROYAL SOC CHEMISTRY

An efficient synthesis of 1-phenyl-2-[(S)-1-aminoethyl]- and 1-phenyl-2-[(S)-1-aminopropyl]-N,N-diethylcyclopropanecarboxamides [1a (PEDC) and 1b (PPDC), respectively], potent NMDA receptor antagonists having a cyclopropane structure, was achieved. We have shown for the first time that C-cyclopropylaldonitrone preferentially exists in the bisected s-trans conformation, due to the characteristic stereoelectronic effects of the cyclopropane ring, by X-ray crystallographic analysis, NMR studies, and theoretical calculations. Based on these findings, the highly stereoselective addition reaction of Grignard reagents to C-cyclopropylaldonitrone 6 was developed, and the reaction was successfully used as the key step for the preparation of the NMDA receptor antagonists 1a and 1b as well as for a newly designed isopropyl-type congener 1c. The facial selectivity of the addition of Grignard reagents can be explained by the attack of the reagents from the less hindered side of the substrate in the predicted bisected s-trans conformation. This Grignard reaction is the first example of a highly stereoselective addition to a nitrone via a non-chelation controlled pathway.

DOI： 10.1039/b008230i

Intracellular Ca2+-mobilizing adenine nucleotides. Synthesis and biological activity of cyclic ADP-carbocyclic-ribose and C-glycosidic analog of adenophostin A

頁： 461-470 2001年

The stereoselective synthesis of 4 '-beta-thioribonucleosides via the Pummerer reaction 査読有り

T Naka, N Minakawa, H Abe, D Kaga, A Matsuda

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 122 巻 ( 30 ) 頁： 7233 - 7243 2000年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

An efficient stereoselective synthesis of 4'-beta-thioribonucleosides 14, 15, 27, and 30 using the Pummerer reaction as the key step is described. The Pummerer reaction of 1,4-anhydro-2-0-(2,4-dimethoxybenzoyl)-3,5-O-( 1,1,3,3-tetraisopropyldisiloxane-1,3-diyl)-3-sulfinyl-D-ribitol (R-10:S-10 = 2.7:1) in the presence of silylated uracil afforded the desired p-anomer of the 4'-thiouridine derivative 11 in 66% yield without formation of its a-anomer. The reaction with R-10 gave 11 in 87% yield, while the one with S-10 resulted in a 27% decrease of the desired product 11 along with a 22% yield of 3,6-O-(1,1,3,3-tetraisopropyldisiloxane-1,3-diyl)-3-hydroxy-2-hydroxymethylthiophene (12). A likely explanation for the observed difference in the reaction of R-10 and S-10 is that the reaction proceeds via an E2 type pathway, which prefers anti elimination. Thus, R-10 would preferentially afford the alpha-thiocarbocation intermediate 21 via an E2 anti elimination under the reaction conditions. The resulting 21 would be expected to react with silylated uracil stereoselectively to give 11 in good yield. However, formation of the more stable tertiary alpha-thiocarbocation intermediate 23, which would prefer to give 12 and/or to decompose, would compete with the formation of the desired 21 in the reaction with S-10. Consequently, this argument would explain the low yields of the desired product 11 and the poor mass balance in the reaction with S-10. When the sulfoxide 10 (R-10:S-10 = >16:1) prepared by oxidation of 9 with ozone was used for the Pummerer reaction, the desired 11 was obtained in 80% yield. Compound 11 was converted to 4'-beta-thiouridine (14) by treatment of 11 with ammonium fluoride, followed by methanolic ammonia. Similarly, 4'-beta-thiocytidine (15) was prepared when silylated N-4-acetylcytosine was used in the Pummerer reaction. For the Pummerer reactions with purine bases, 6-chloropurine and 2-amino-6-chloropurine were found to be the most suitable. When the reactions were conducted in a mixture of acetonitrile and 1,2-dichloroethane at room temperature, followed by reflux, the desired products 25 and 28 were obtained in 65% and 56% yields, respectively. These compounds were then converted to 4'-beta-thioadenosine (27) and 4'-beta-thioguanosine (30) under the usual conditions. This is therefore the first time that the stereoselective synthesis of 4'-beta-thioribonucleosides has been performed using the neighboring group participation of the Pummerer reaction.

DOI： 10.1021/ja000541o

Synthesis of the C-glycosidic analogue of adenophostin A and its uracil congener as potential IP3 receptor ligands. Stereoselective construction of the C-glycosidic structure by a temporary silicon-tethered radical coupling reaction 査読有り

H Abe, S Shuto, A Matsuda

JOURNAL OF ORGANIC CHEMISTRY 65 巻 ( 14 ) 頁： 4315 - 4325 2000年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

Synthesis of the C-glycosidic analogue 9 of adenophostin A, a very potent IP3 receptor agonist, and its uracil congener 10 was achieved via a temporary silicon-tethered radical coupling reaction as the key step. Phenyl 3,4,6-tri-O-(p-methoxybenzyl)-1-seleno-beta-D-glucopyranoside (27) and 3-deoxy3-methylene-1,2-O-isopropylidene-alpha-D-erythro (30) were connected by a dimethylsilyl tether to give the radical coupling reaction substrate 24, which was successively treated with Bu-3-SnH/AIBN in benzene and TBAF in THF to give the coupling product 25 with the desired (3a,1'a)-configuration as the major product. From 25, the targets 9 and 10 were synthesized via introduction of adenine or uracil base by Vorbruggen's method and phosphorylation of the hydroxyls by the phosphoramidite method.

DOI： 10.1021/jo0001333

Stereoselective synthesis of alpha and beta-C-glucosides via radical cyclization with an allylsilyl tether. Control of the stereoselectivity by changing the conformation of the pyranose ring 査読有り

S Shuto, M Terauchi, Y Yahiro, H Abe, S Ichikawa, A Matsuda

TETRAHEDRON LETTERS 41 巻 ( 21 ) 頁： 4151 - 4155 2000年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

An efficient method for preparing both 1 alpha- and 1 beta-C-glucosides having a 3-hydroxypropyl group at the anomeric position via a radical cyclization reaction with an allylsilyl tether was developed. The stereoselectivity of the radical cyclization can be controlled by the conformation of the pyranose ring, which is effectively manipulated by the hyroxyl protecting groups. (C) 2000 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0040-4039(00)00556-6

Synthesis of the C-glycosidic analog of adenophostin A, a potent IP3 receptor agonist, using a temporary silicon-tethered radical coupling reaction as the key step 査読有り

H Abe, S Shuto, A Matsuda

TETRAHEDRON LETTERS 41 巻 ( 14 ) 頁： 2391 - 2394 2000年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

Synthesis of the C-glycosidic analog (3) of adenophostin A, a very potent IP3 receptor agonist, was achieved using a temporary silicon-tethered reductive radical coupling reaction as the key step. Radical reaction of the silaketal substrate 6 with Bu3SnH/AIBN in benzene occurred stereoselectively, and subsequent desilylation gave the desired C-glycosidic disaccharide 7 with the (3 alpha,1'alpha)-configuration as the major product. Compound 7 was converted into the target 3 via the introduction of an adenine base by a Vorbruggen glycosylation reaction. (C) 2000 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0040-4039(00)00171-4

Mechanistic study of the ring-enlargement reaction of (3-oxa-2-silacyclopentyl)methyl radicals into 4-oxa-3-silacyclohexyl radicals. Evidence for a pentavalent silicon-bridging radical transition state in 1,2-rearrangement reactions of beta-silyl radicals 査読有り

S Shuto, Sugimoto, I, H Abe, A Matsuda

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 122 巻 ( 7 ) 頁： 1343 - 1351 2000年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

A mechanistic study was performed on a novel radical ring-enlargement reaction of (3-oxa-2-silacyclopentyl)methyl radicals into 4-oxa-3-silacyclohexyl radicals. Two pathways, one via a pentavalent silicon bridging radical transition state (or intermediate), the other via beta-elimination to give a ring-opened silyl radical, can be postulated. The radical reactions of 1 and 2, which are precursors for a (3-oxa-2-silacyclopentyl)methyl radical C' and a 4-oxa-3-silacyclohexyl radical D', respectively, showed that the ring-enlargement rearrangement of C' into D' is irreversible. H-1 NMR analysis of the radical reactions of 8a and 8b, which have an asymmetric center at silicon, indicated that the configuration at the silicon atom is retained via a pentavalent silicon-bridging radical transition state (or intermediate) during the ring-enlargement reaction. Furthermore, examination of the radical ring-enlargement reaction with a deuterium-labeled substrate 12D showed that the ring-enlargement reaction did not involve beta-elimination to give a ring-opened silyl radical. Based on these results, we conclude that the ring-enlargement reaction occurs via a pentavalent silicon-bridging radical transition state (or intermediate). This is the first experimental evidence for such a pentavalent silicon radical, which has been previously postulated to understand radical reactions of organic silicon compounds.

DOI： 10.1021/ja993239s

Conjugation of dipeptide to fluorescent dyes enhances its affinity for a dipeptide transporter (PEPT1) in human intestinal Caco-2 cells 査読有り

H Abe, M Satoh, S Miyauchi, S Shuto, A Matsuda, N Kamo

BIOCONJUGATE CHEMISTRY 10 巻 ( 1 ) 頁： 24 - 31 1999年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER CHEMICAL SOC

Dipeptide transporters in small intestine have a very wide substrate specificity, so that the transporter sometimes serves as a carrier for peptide-like compounds. We have synthesized dipeptide analogues conjugated at an E-amino group of Lys in Val-Lys or Lys-Sar with fluorescent compounds such as fluorescein isothiocyanate and coumarin-3-carboxylic acid. Uptakes of these peptide analogues were examined by measuring intracellular accumulations into monolayers of the human intestinal epithelial cell line Caco-2 expressing the dipeptide transporter PEPT1. Kinetic analysis and effects of addition either of uncoupler (protonophore) or by Gly-Sar, one of the good substrates of PEPT1, revealed that fluorescent dipeptides were taken up by passive diffusion. In contrast, these analogues remarkably inhibited the Gly-Sar uptake by Caco-2 cells. Among the fluorescent analogues synthesized in this paper, Vaf-Lys(Flu) was the most potent competitive inhibitor against the Gly-Sar uptake with an inhibition constant of 5 mu M. This value is the smallest among those ever reported: Val-Lys(Flu) has the highest affinity for PEPT1 among chemicals ever reported. The importance of the hydrophobic part of the substrate was pointed out.

DOI： 10.1021/bc980049i

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