Faculty Profiles - ABE Hiroshi

PubMed

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

Structure Formation and Catalytic Activity of DNA Dissolved in Organic Solvents Reviewed

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

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION Vol. 51 ( 26 ) page： 6475 - 6479 2012

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

INTERNATIONAL JOURNAL OF NANOMEDICINE Vol. 7 page： 5437 - 5450 2012

Language：English Publishing type：Research paper (scientific journal) Publisher：DOVE MEDICAL PRESS LTD

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.

DOI： 10.2147/IJN.S36350

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

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 Vol. 22 ( 10 ) page： 2082 - 92 2011.10

Synthesis, Structure, and Biological Activity of Dumbbell-Shaped Nanocircular RNAs for RNA Interference Reviewed

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

BIOCONJUGATE CHEMISTRY Vol. 22 ( 10 ) page： 2082 - 2092 2011.10

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

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 the most promising design for in vivo applications for RNA medicine.

DOI： 10.1021/bc203154

Synthesis and Characterization of a Series of Highly Fluorogenic Substrates for Glutathione Transferases, a General Strategy Reviewed

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

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY Vol. 133 ( 35 ) page： 14109 - 14119 2011.9

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

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.

DOI： 10.1021/ja205500y

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

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

ACS MEDICINAL CHEMISTRY LETTERS Vol. 2 ( 9 ) page： 692 - 697 2011.9

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

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 beta-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).

DOI： 10.1021/ml2001054

PEGylated antibody in organic media Reviewed

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

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 111 ( 5 ) page： 564 - 568 2011.5

Language：English Publishing type：Research paper (scientific journal) Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN

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. (C) 2011, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2011.01.001

Indomethacin analogues that enhance doxorubicin cytotoxicity in multidrug resistant cells without cox inhibitory activity Reviewed

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

ACS Medicinal Chemistry Letters Vol. 2 ( 5 ) page： 353 - 357 2011.5

Language：English Publishing type：Research paper (scientific journal)

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 Reviewed

Mingzhe Liu, Hiroshi Abe, Yoshihiro Ito

KOBUNSHI RONBUNSHU Vol. 68 ( 7 ) page： 405 - 416 2011

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：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 Reviewed

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

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION Vol. 50 ( 50 ) page： 12020 - 12023 2011

Synthesis and characterization of small circular double-stranded RNAs Reviewed

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

CHEMICAL COMMUNICATIONS Vol. 47 ( 7 ) page： 2125 - 2127 2011

Language：English Publishing type：Research paper (scientific journal) Publisher：ROYAL SOC CHEMISTRY

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.

DOI： 10.1039/c0cc04551a

Branched RNA nanostructures for RNA interference Reviewed

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

CHEMICAL COMMUNICATIONS Vol. 47 ( 29 ) page： 8367 - 8369 2011

Language：English Publishing type：Research paper (scientific journal) Publisher：ROYAL SOC CHEMISTRY

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.

DOI： 10.1039/c1cc11780g

Characterization of New Potential Anticancer Drugs Designed To Overcome Glutathione Transferase Mediated Resistance Reviewed

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

MOLECULAR PHARMACEUTICS Vol. 8 ( 5 ) page： 1698 - 1708 2011

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

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 add, 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.

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 Vol. 68 ( 7 ) page： 405-416 2011

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

Liu Mingzhe, Abe Hiroshi, Ito Yoshihiro

KOBUNSHI RONBUNSHU Vol. 68 ( 7 ) page： 405 - 416 2011

Publisher：Kobunshi Ronbunshu

DOI： 10.1295/koron.68.405

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

Yasuyuki Hirama, Hiroshi Abe, Noriaki Minakawa, Akira Matsuda

TETRAHEDRON Vol. 66 ( 43 ) page： 8402 - 8406 2010.10

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

We describe herein the synthesis and properties of the novel nucleoside derivative, 4,7-diamino-2-(2-deoxy-beta-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-beta-D-erythro-pentofuranosyl) pyrrolo[2,3-d]pyrimidine (9) with acrylonitrile afforded 2,4-diamino-5-[(E)-1-cyano-2-ethenyl]-7-(2-deoxy-beta-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-(2deoxy-beta-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. (C) 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 Vol. 1043 page： 111 - 123 2010.8

Publisher：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