Updated on 2024/10/03

写真a

 
NAKAO, Yoichi
 
Affiliation
Faculty of Science and Engineering, School of Advanced Science and Engineering
Job title
Professor
Degree
博士(農学) ( 東京大学 )

Research Experience

  • 2012.04
    -
    Now

    Waseda University

  • 2007.04
    -
    2012.03

    Waseda University   Faculty of Science and Engineering

  • 2004
    -
    2007.03

    The University of Tokyo   Graduate School of Agricultural and Life Sciences

  • 1996.06
    -
    2004

    The University of Tokyo

  • 1994.07
    -
    1996.09

    University of Hawai'i   Dept. Chemistry   posdoc

Education Background

  • 1991.04
    -
    1994.03

    The University of Tokyo  

  • 1989.04
    -
    1991.03

    The University of Tokyo  

  • 1985.04
    -
    1989.03

    The University of Tokyo  

Committee Memberships

  • 2011
    -
    Now

    天然物討論会  世話人

  • 2010
    -
    Now

    日本ケミカルバイオロジー学会  世話人

  • 2007
    -
    Now

    化学生態学研究会  世話人

  • 2012
    -
    2014

    日本化学会  関東支部幹事

  • 2012
    -
    2013

    日本化学会  代議員

  • 1997
    -
    2006

    天然物談話会  世話人

  •  
     
     

    日本水産学会  庶務幹事

▼display all

Professional Memberships

  • 1995
    -
    Now

    American Chemical Society

  •  
     
     

    日本バイオインフォマティクス学会

  •  
     
     

    日本寄生虫学会

  •  
     
     

    日本生物工学会

  •  
     
     

    日本化学会

  •  
     
     

    日本ケミカルバイオロジー学会

  •  
     
     

    日本農芸化学会

  •  
     
     

    日本分子生物学会

  •  
     
     

    日本水産学会

  •  
     
     

    日本エピジェネティクス研究会

  •  
     
     

    日本がん分子標的治療学会

▼display all

Research Areas

  • Food sciences   functional food chemistry / Bioorganic chemistry   natral products chemistry, chemical biology

Research Interests

  • functional food chemistry

  • differentiation control of stem cells

  • epigenetics

  • natural products chemistry

  • chemical biology

Media Coverage

  • しなやかに生きる 94歳、辰巳芳子さんからの伝言。生ハムづくりを通して学んだ、生きること、考えること。

    Newspaper, magazine

    2019.08

  • 教室は「10年待ち」! 93歳料理研究家 いのちのスープ

    TV or radio program

    2018.09

  • 未来の起源

    TBS  

    2017.04

  • 食の未来対談 「食と人間の関係」が、科学的に解明されつつあります

    Newspaper, magazine

    2017.04

  • 万能細胞から心筋細胞作製 海洋生物の抽出物 効果 早大、低コスト育成見込む

    Newspaper, magazine

    2017.02

  • 万能細胞の試薬 神経へ効率分化 早大など開発

    Newspaper, magazine

    2015.12

  • 『大学 知の明日を築く』早稲田大東日本大震災復興研究拠点 防災・医療知識 海外に

    Newspaper, magazine

    2013.05

  • 海中生物から抗原虫薬 リーシュマニア症 安価で効率高く 早大などが候補発見

    Newspaper, magazine

    2012.01

  • 『命のゆりかご 瀬戸内の多様な生態系』15 親薬の源 毒の成分 研究者が熱視線

    Newspaper, magazine

    2011.05

  • 天然化合物で細胞増殖 副作用がない薬の開発へ

    Newspaper, magazine

    2011.03

  • 万能細胞から心筋 新技術で効率良く

    Newspaper, magazine

    2011.03

▼display all

 

Papers

  • Aurantoside L, a New Tetramic Acid Glycoside with Anti-Leishmanial Activity Isolated from the Marine Sponge Siliquariaspongia japonica

    Yasumoto Oyadomari, Yasuyuki Goto, Keisuke Suganuma, Shin-ichiro Kawazu, Leontine Becking, Nobuhiro Fusetani, YOICHI NAKAO

    Marine Drugs    2024.04  [Refereed]

    DOI

    Scopus

  • Two Onnamide Analogs from the Marine Sponge Theonella conica: Evaluation of Geometric Effects in the Polyene Systems on Biological Activity

    Fumiaki Nakamura, Hiroshi Kimura, Nobuhiro Fusetani, Yoichi Nakao

    Molecules    2023.03

    DOI

    Scopus

    1
    Citation
    (Scopus)
  • Cultivation of previously uncultured sponge-associated bacteria using advanced cultivation techniques: A perspective on possible key mechanisms

    Dawoon Jung, Koshi Machida, Yoichi Nakao, Jeffrey S. Owen, Shan He, Tomonori Kindaichi, Akiyoshi Ohashi, Yoshiteru Aoi

    Frontiers in Marine Science   9  2022.08

     View Summary

    Most of the microbes from natural habitats cannot be cultivated with standard cultivation in laboratory, and sponge-associated microbes are no exception. We used two advanced methods based on a continuous-flow bioreactor (CF) and in situ cultivation (I-tip) to isolate previously uncultivated marine sponge-associated bacteria. We also characterized the physiological properties of the isolates from each method and attempted to clarify the mechanisms operating in each cultivation method. A greater number of novel bacteria were isolated using CF and in situ cultivation compared to standard direct plating (SDP) cultivation. Most isolates from CF cultivation were poor growers (with lower specific growth rates and saturated cell densities than those of isolates from SDP cultivation), demonstrating that it is effective to carry out pre-enrichment cultivation targeting bacteria that are less competitive on conventional cultivation, especially K-strategists and bacterial types inhibited by their own growth. Isolates from in situ cultivation showed a positive influence on cell recovery stimulated by chemical compounds in the extract of sponge tissue, indicating that some of the bacteria require a “growth initiation factor” that is present in the natural environment. Each advanced cultivation method has its own distinct key mechanisms allowing cultivation of physiologically and phylogenetically different fastidious bacteria for cultivation compared with conventional methods.

    DOI

    Scopus

    4
    Citation
    (Scopus)
  • Visualization of the dynamic interaction between nucleosomal histone H3K9 tri-methylation and HP1α chromodomain in living cells

    Kazuki Sasaki, Michihiro Suzuki, Takeshi Sonoda, Tilman Schneider-Poetsch, Akihiro Ito, Motoki Takagi, Shinya Fujishiro, Yoshihiro Sohtome, Kosuke Dodo, Takashi Umehara, Hiroyuki Aburatani, Kazuo Shin-ya, Yoichi Nakao, Mikiko Sodeoka, Minoru Yoshida

    Cell Chemical Biology    2022.06

    DOI

    Scopus

    4
    Citation
    (Scopus)
  • Coronarin D, a Metabolite from the Wild Turmeric, Curcuma aromatica, Promotes the Differentiation of Neural Stem Cells into Astrocytes.

    Satoshi Otsuka, Midori Kawamura, Shutaro Fujino, Fumiaki Nakamura, Daisuke Arai, Nobuhiro Fusetani, Yoichi Nakao

    Journal of agricultural and food chemistry   70 ( 10 ) 3300 - 3309  2022.03  [International journal]

     View Summary

    Plants in the genus Curcuma have been widely used as traditional medicines in Asian countries. These plants contain bioactive compounds with neuroprotective properties or activities that increase neural stem cells (NSCs) and neurons. However, bioactive components in Curcuma that promote the differentiation of NSCs into astrocytes have not yet been reported. Here, the effects of Curcuma extracts on the in vitro differentiation of embryonic stem-cell-derived NSCs were evaluated. The extract of the wild turmeric, Curcuma aromatica, strongly promoted the differentiation of NSCs into astrocytes. Bioassay-guided isolation yielded coronarins C (1) and D (2), as well as (E)-labda-8(17),12-diene-15,16-dial (3) as the bioactive compounds. Coronarin D (2) markedly promoted the differentiation of NSCs into astrocytes up to approximately 4 times (3.64 ± 0.48) and increased the expression level of GFAP at the mRNA and protein level, while compounds 1 and 3 exhibited only weak effects, suggesting that the 15-hydroxy-Δ12-γ-lactone moiety is important for bioactivity. Moreover, compound 2 increased the number of pSTAT3-positive cells, suggesting that compound 2 promoted astrocytic differentiation through JAK/STAT signaling pathway.

    DOI PubMed

    Scopus

    7
    Citation
    (Scopus)
  • Mutualistic relationship between Nitrospira and concomitant heterotrophs.

    Chiho Murakami, Koshi Machida, Yoichi Nakao, Tomonori Kindaichi, Akiyoshi Ohashi, Yoshiteru Aoi

    Environmental microbiology reports   14 ( 1 ) 130 - 137  2022.02  [International journal]

     View Summary

    Nitrifying chemoautotrophs support the growth of diverse concomitant heterotrophs in natural or engineered environments by supplying organic compounds. In this study, we aimed to investigate this microbial association, especially (i) to distinguish whether the relationship between nitrifying chemoautotrophs and heterotrophs is commensal or mutualistic, and (ii) to clarify how heterotrophs promote the growth of autotrophic nitrite-oxidizing bacteria (Nitrospira). Pure cultured Nitrospira (Nitrospira sp. ND1) was employed in this study. Heterotrophs growing with metabolic by-products of Nitrospira as a sole carbon source were isolated from several environmental samples and used to test the growth-promoting activity of Nitrospira. Furthermore, liquid chromatography-mass spectrometry analysis was conducted to evaluate how heterotrophs consumed chemical compounds produced by Nitrospira and newly produced during co-cultivation. Notably, Nitrospira growth was stimulated by co-cultivation with some heterotrophs and the addition of spent media of some strains, suggesting that not only heterotrophs but also Nitrospira received benefits from their mutual co-existence. Furthermore, the data suggested that some of the growth-promoting heterotrophs provided as-yet-unidentified growth-promoting factors to Nitrospira. Overall, Nitrospira and heterotrophs thus appear to exhibit a mutualistic relationship. Such mutualistic relationships between autotrophs and heterotrophs would contribute to the stability and diversity of microbial ecosystems.

    DOI PubMed

    Scopus

    5
    Citation
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  • Efficient biallelic knock-in in mouse embryonic stem cells by in vivo-linearization of donor and transient inhibition of DNA Polymerase θ/DNA-PK

    Daisuke Arai, Yoichi Nakao

       2021.05

     View Summary

    CRISPR/Cas9-mediated homology-directed repair (HDR) is used for error-free targeted knock-in of foreign donor DNA. However, the low efficiency of HDR-mediated knock-in hinders establishment of knock-in clones. Double-strand breaks (DSBs) induced by CRISPR/Cas9 are preferentially repaired by non-homologous end joining (NHEJ) or microhomology-mediated end joining (MMEJ) before HDR can occur, thereby preventing HDR-mediated knock-in. NHEJ/MMEJ also cause random integrations, which give rise to false-positive knock-in events, or silently disrupt the genome. In this study, we optimized an HDR-mediated knock-in method for mouse embryonic stem cells (mESCs). We succeeded in improving efficiency of HDR-mediated knock-in of a plasmid donor while almost completely suppressing NHEJ/MMEJ-based integration by combining in vivo-linearization of the donor plasmid, transient knockdown of DNA Polymerase θ, and chemical inhibition of DNA-dependent protein kinase (DNA-PK) by M3814. This method also dramatically improved the efficiency of biallelic knock-in; at the Rosa26a locus, 95% of HDR-mediated knock-in clones were biallelic. We designate this method BiPoD (Biallelic knock-in assisted by Pol θ and DNA-PK inhibition). BiPoD achieved simultaneous efficient biallelic knock-in into two loci. BiPoD, therefore, enables rapid and easy establishment of biallelic knock-in mESC lines.

    DOI

  • Disrupting tumor onset and growth via selective cell tagging (SeCT) therapy.

    Kenward Vong, Tsuyoshi Tahara, Sayaka Urano, Igor Nasibullin, Kazuki Tsubokura, Yoichi Nakao, Almira Kurbangalieva, Hirotaka Onoe, Yasuyoshi Watanabe, Katsunori Tanaka

    Science advances   7 ( 17 )  2021.04  [International journal]

     View Summary

    This study presents the early framework of selective cell tagging (SeCT) therapy, which is the concept of preferentially labeling specific cells in vivo with chemical moieties that can elicit a therapeutic response. Using glycosylated artificial metalloenzyme (GArM)-based protein labeling, this study reports two separate functional strategies. In one approach, early tumor onset can be suppressed by tagging cancer cells in living mice with an integrin-blocking cyclic-Arg-Gly-Asp (cRGD) moiety, thereby disrupting cell adhesion onto the extracellular matrix. In another approach, tumor growth in mice can be reduced by tagging with a cytotoxic doxorubicin moiety. Subsequent cell death occurs following internalization and drug release. Overall, experiments have shown that mouse populations receiving the mixture of SeCT labeling reagents exhibited a significant delay/reduction in tumor onset and growth compared with controls. Highlighting its adaptability, this work represents a foundational step for further development of SeCT therapy and its potential therapeutic applications.

    DOI PubMed

    Scopus

    12
    Citation
    (Scopus)
  • Small-Scale Preparation of Fluorescently Labeled Chemical Probes from Marine Cyclic Peptides, Kapakahines A and F

    Rie Kamihira, Yoichi Nakao

    Marine Drugs   19 ( 2 ) 76 - 76  2021.01  [Refereed]  [Invited]

    Authorship:Last author, Corresponding author

     View Summary

    A number of bioactive marine natural products have been isolated so far, but it is still difficult to disclose their modes of action. In this study, we prepared fluorescently labeled chemical probes from the cytotoxic marine cyclic peptides kapakahines A (1) and F (2) to visualize their localization as the first step of the study of their modes of action. We used fluorescent dyes 3a or 3a/b (a 1:1 mixture of 3a and 3b) whose terminal N-hydroxysuccinimide (NHS) group can react with the free amino groups of kapakahines. The fluorescently labeled kapakahine A (Kap A-5-FL, 5a) stained P388 murine leukemia cells and HeLa human cervical cancer cells, while cells treated with fluorescently labeled kapakahine F (Kap F-5-FL, 6a) only weakly stained them. Further analysis of the confocal images of the stained cells with higher magnification (×100) indicated the localization of Kap A-5-FL (5a) in the cells. In this paper, we report the small-scale preparation and a new delivery method of fluorescent probes, as well as the application of these procedures to cell staining.

    DOI

  • Triggering Growth via Growth Initiation Factors in Nature: A Putative Mechanism for in situ Cultivation of Previously Uncultivated Microorganisms.

    Dawoon Jung, Koshi Machida, Yoichi Nakao, Tomonori Kindaichi, Akiyoshi Ohashi, Yoshiteru Aoi

    Frontiers in microbiology   12   537194 - 537194  2021  [International journal]

     View Summary

    Most microorganisms resist cultivation under standard laboratory conditions. On the other hand, cultivating microbes in a membrane-bound device incubated in nature (in situ cultivation) can be an effective approach to overcome this limitation. In the present study, we applied in situ cultivation to isolate diverse previously uncultivated marine sponge-associated microbes and comparatively analyzed this method's efficiencies with those of the conventional method. Then, we attempted to investigate the key and previously unidentified mechanism of growing uncultivated microorganisms by in situ cultivation focusing on growth triggering via growth initiation factor. Significantly more novel and diverse microbial types were isolated via in situ cultivation than by standard direct plating (SDP). We hypothesized that some of environmental microorganisms which resist cultivation are in a non-growing state and require growth initiation factors for the recovery and that these can be provided from the environment (in this study from marine sponge). According to the hypothesis, the effect of the sponge extract on recovery on agar medium was compared between strains derived from in situ and SDP cultivation. Adding small amounts of the sponge extracts to the medium elevated the colony-formation efficiencies of the in situ strains at the starvation recovery step, while it showed no positive effect on that of SDP strains. Conversely, specific growth rates or saturated cell densities of all tested strains were not positively affected. These results indicate that, (1) the sponge extract contains chemical compounds that facilitate recovery of non-growing microbes, (2) these substances worked on the in situ strains, and (3) growth initiation factor in the sponge extract did not continuously promote growth activity but worked as triggers for regrowth (resuscitation from non-growing state).

    DOI PubMed

    Scopus

    12
    Citation
    (Scopus)
  • Histone modification dynamics as revealed by multicolor immunofluorescence-based single-cell analysis

    Yoko Hayashi-Takanaka, Yuto Kina, Fumiaki Nakamura, Leontine E. Becking, Yoichi Nakao, Takahiro Nagase, Naohito Nozaki, Hiroshi Kimura

    Journal of Cell Science   133 ( 14 ) jcs243444 - jcs243444  2020.07  [International journal]

     View Summary

    <title>ABSTRACT</title>Post-translational modifications on histones can be stable epigenetic marks or transient signals that can occur in response to internal and external stimuli. Levels of histone modifications fluctuate during the cell cycle and vary among different cell types. Here, we describe a simple system to monitor the levels of multiple histone modifications in single cells by multicolor immunofluorescence using directly labeled modification-specific antibodies. We analyzed histone H3 and H4 modifications during the cell cycle. Levels of active marks, such as acetylation and H3K4 methylation, were increased during the S phase, in association with chromatin duplication. By contrast, levels of some repressive modifications gradually increased during G2 and the next G1 phases. We applied this method to validate the target modifications of various histone demethylases in cells using a transient overexpression system. In extracts of marine organisms, we also screened chemical compounds that affect histone modifications and identified psammaplin A, which was previously reported to inhibit histone deacetylases. Thus, the method presented here is a powerful and convenient tool for analyzing the changes in histone modifications.

    DOI PubMed

    Scopus

    20
    Citation
    (Scopus)
  • Combinatorial Effects of Soluble, Insoluble, and Organic Extracts from Jerusalem Artichokes on Gut Microbiota in Mice

    Hiroyuki Sasaki, Yijin Lyu, Yuki Nakayama, Fumiaki Nakamura, Aya Watanabe, Hiroki Miyakawa, Yoichi Nakao, Shigenobu Shibata

    Microorganisms   8 ( 6 ) 954 - 954  2020.06  [International journal]

     View Summary

    Jerusalem artichokes contain high amounts of inulin, which is a prebiotic that supports digestive health, as well as a variety of insoluble fibers and caffeoylquinic acid. The individual impact of these components on gut microbiota is well known; however, the combinatorial effects are less clear. In this investigation, we fractionated Jerusalem artichokes into three parts (water-soluble extract, insoluble extract, and organic extract) and powdered them. Mice were fed a high-fat diet that included one or more of these extracts for 10 days, and then their cecal pH, cecal short-chain fatty acids (SCFAs), and fecal microbiota were evaluated. The combination of the water-soluble and organic extract decreased cecal pH and increased the concentration of SCFAs and led to dynamic changes in the composition of the gut microbiota. These results demonstrate that both the water-soluble and organic extracts in Jerusalem artichokes are bioactive substances that are capable of changing SCFA production and the composition of gut microbiota. Powdered Jerusalem artichokes, rather than inulin supplements, may be superior for promoting a healthy gut.

    DOI PubMed

    Scopus

    10
    Citation
    (Scopus)
  • Total Synthesis and Biological Evaluation of Kakeromamide A and Its Analogues

    Meng Zhao, Yi Xiao, Satoshi Otsuka, Yoichi Nakao, Yian Guo, Tao Ye

    Frontiers in Chemistry   8   410 - 410  2020.05  [International journal]

     View Summary

    Kakeromamide A (1), the first marine cyclopeptide inducing neural stem cells differentiation into astrocytes, was synthesized in 12 longest linear steps and 14% overall yield. Using this synthetic approach, four analogs of kakeromamide A were prepared and evaluated for neural differentiation- modulating activity.

    DOI PubMed

    Scopus

    4
    Citation
    (Scopus)
  • Efficient synthesis ofmarine alkaloid ageladine A and its structural modification for exploring new biological activity

    Takayuki Iwata, Koichi Fukase, Yoichi Nakao, Katsunori Tanaka

    Yuki Gosei Kagaku Kyokaishi/Journal of Synthetic Organic Chemistry   78 ( 1 ) 51 - 59  2020

     View Summary

    Natural products have been regarded as a pivotal source for biological active compounds. In addition, structural modification of natural products has been simultaneously received much attention because these modified natural products would show not only enhanced activity and selectivity, but possibly also an entirely distinct bioactivity from those of the parent natural products. We describe herein a series of study on structual modification of ageladine A for exploring new biological activity. Ageladine A was isolated from the marine sponge Agelas nakamurai as an antiangiogenic compound. Based on total syntheses of this alkalod, its structual modifications were caried out as a part of SAR study, which revealed that a pyridine derivative shows a more potent activity. More recently, we have developed an efficient bio-inspired cascade reaction for the synthesis of ageladine A The cascade reaction involves an aza-electrocyclization and a novel 2-aminoimidazole formation that is modeled after a post-translational modification of arginine residue in protein. Employing the cascade reaction, ageladine A and its N1-substituted analogues were efficiently synthesized in one-pot fashion from anilines or guanidines. In addition, it was found that some analogs showed significant activity on modulating neuronal differentiation. Namely, these analogs selectively activate or inhibit the differentiation of neural stem cells to neurons, while negligible in astrocyte differentiation. The series of study represent a successful case in altering native bioactivity of natural products by structural modification.

    DOI

    Scopus

    2
    Citation
    (Scopus)
  • Efficient route to RIKEN click probes for glycoconjugation.

    Nakamura, K, Tsubokura, K, Kurbangalieva, A, Nakao, Y, Murase, T, Shimoda, T, Tanaka, K

    J. Carbohyd. Chem.    2019  [Refereed]

    DOI

  • Effect of mycalolides isolated from a marine sponge Mycale aff. nullarosette on actin in living cells .

    Hayashi-Takanaka, Y, Kina, Y, Nakamura, F, Yamazaki, S, Harata, M, van Soest, R. W. M, Kimura, H, Nakao, Y

    Sci. Rep.   9 ( 1 ) 7540 - 7540  2019  [Refereed]  [International journal]

     View Summary

    Discovery of novel bioactive compounds is important not only for therapeutic purposes but also for understanding the mechanisms of biological processes. To screen bioactive compounds that affect nuclear morphology in marine organism extracts, we employed a microscopy-based assay using DNA staining of human cancer cells. A crude extract from a marine sponge Mycale aff. nullarosette, collected from the east coast of Japan, induced cellular binucleation. Fractionation of the extract led to the isolation of mycalolides A and B, and 38-hydroxymycalolide B as the active components. Mycalolides have been identified as marine toxins that induce depolymerization of the actin filament. Live cell imaging revealed that low concentrations of mycalolide A produce binucleated cells by inhibiting the completion of cytokinesis. At higher concentrations, however, mycalolide A causes immediate disruption of actin filaments and changes in cell morphology, yielding rounded cells. These results suggest that the completion of cytokinesis is a process requiring high actin polymerization activity. Furthermore, luciferase reporter assays with mycalolide A treatments support the view that the level of globular actin can affect transcription of a serum response gene.

    DOI PubMed

    Scopus

    8
    Citation
    (Scopus)
  • Correction to: Halistanol sulfates I and J, new SIRT1-3 inhibitory steroid sulfates from a marine sponge of the genus Halichondria.

    Fumiaki Nakamura, Norio Kudo, Yuki Tomachi, Akiko Nakata, Misao Takemoto, Akihiro Ito, Hodaka Tabei, Daisuke Arai, Nicole de Voogd, Minoru Yoshida, Yoichi Nakao, Nobuhiro Fusetani

    The Journal of antibiotics   71 ( 4 ) 483 - 483  2018.03  [Domestic journal]

     View Summary

    The original article can be found online at https://doi.org/10.1038/ja.2017.145 .

    DOI PubMed

  • Halistanol sulfates i and J, new SIRT1-3 inhibitory steroid sulfates from a marine sponge of the genus Halichondria

    Fumiaki Nakamura, Norio Kudo, Yuki Tomachi, Akiko Nakata, Misao Takemoto, Akihiro Ito, Hodaka Tabei, Daisuke Arai, Nicole De Voogd, Minoru Yoshida, Yoichi Nakao, Nobuhiro Fusetani

    Journal of Antibiotics   71 ( 2 ) 273 - 278  2018.02  [Refereed]

    Authorship:Corresponding author

     View Summary

    Two new analogs of halistanol sulfate (1) were isolated from a marine sponge Halichondria sp. collected at Hachijo-jima Island. Structures of these new halistanol sulfates I (2) and J (3) were elucidated by spectral analyses. Compounds 1-3 showed inhibitory activity against SIRT 1-3 with IC 50 ranges of 45.9-67.9, 18.9-21.1 and 21.8-37.5 μM, respectively. X-ray crystallography of the halistanol sulfate (1) and SIRT3 complex clearly indicates that 1 binds to the exosite of SIRT3 that we have discovered in this study.

    DOI

    Scopus

    19
    Citation
    (Scopus)
  • Kakeromamide A, a new cyclic pentapeptide inducing astrocyte differentiation isolated from the marine cyanobacterium Moorea bouillonii.

    Nakamura, F, Maejima, H, Kawamura, M, Arai, D, Okino, T, Zhao, M, Ye, T, Lee, J, Chang, Y.-T, Fusetan, N, Nakao, Y

    Bioorg. Med. Chem. Lett.   28   2206 - 2209  2018  [Refereed]

    Authorship:Last author, Corresponding author

    DOI

  • Sameuramide A, a new cyclic depsipeptide isolated from an ascidian of the family Didemnidae.

    Machida, K, Arai, D, Katsumata, R, Otsuka, S, Yamashita, J. K, Ye, T, Tang, S, Fusetani, N, Nakao, Y

    Bioorg. Med. Chem.   26 ( 13 ) 3852 - 3857  2018  [Refereed]  [International journal]

    Authorship:Last author, Corresponding author

     View Summary

    Sameuramide A (1), a new cyclic depsipeptide encompassing one each of alanine, N-methyl alanine, N-methyl dehydroalanine, N,O-dimethyl threonine, phenyllactic acid, three β-hydroxy leucines, and two propionates, was isolated from a didemnid ascidian collected at the northern part of Japan. The planar structure was established based on the interpretation of MS and NMR data. The absolute configuration of the subunits was determined by the advanced Marfey's method and the chiral LC-MS analysis. Compound 1 exhibited the activity of maintaining colony formation of murine embryonic stem (mES) cells without leukemia inhibitory factor (LIF). Down regulation of the gene expression of Krüppel-like transcription factor 4 (Klf4) indicated that 1 itself was not able to maintain the undifferentiated state of the mES cells. However, the expression levels of the marker genes (Nestin, T, Sox17) for three germ layers were upregulated in embryoid bodies (EBs) after treatment of 1 together with LIF, suggesting that 1 plays a supportive role for LIF in maintaining the multipotency of mES cells.

    DOI PubMed

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    11
    Citation
    (Scopus)
  • Dolabellol A, a New Halogenated Diterpene Isolated from the Opisthobranch Dolabella auricularia

    Koshi Machida, Takuro Matsumoto, Nobuhiro Fusetani, Yoichi Nakao

    CHEMISTRY LETTERS   46 ( 11 ) 1676 - 1678  2017.09  [Refereed]

     View Summary

    A new halogenated diterpene, dolabellol A (1), was isolated from the opisthobranch Dolabella auricularia. The planar structure of dolabellol A was elucidated by NMR spectroscopic analysis and chemical reactions. The absolute stereochemistry was determined by single-crystal X-ray diffraction studies.

    DOI

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    6
    Citation
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  • Cancer cell targeting driven by selective polyamine reactivity with glycine propargyl esters

    Kenward K. H. Vong, Kazuki Tsubokura, Yoichi Nakao, Tomonori Tanei, Shinzaburo Noguchi, Shinobu Kitazume, Naoyuki Taniguchi, Katsunori Tanaka

    CHEMICAL COMMUNICATIONS   53 ( 60 ) 8403 - 8406  2017.08  [Refereed]

     View Summary

    Rapidly growing cancer cells have increased levels of intracellular polyamines compared to normal, healthy tissues. Based on the selective reactivity of glycine propargyl esters, probes were synthesized that show evidence for selective polyamine reactivity, which was then applied for selective cancer cell imaging studies.

    DOI

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    10
    Citation
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  • Identification of zinc finger transcription factor EGR2 as a novel acetylated protein

    Kota Noritsugu, Akihiro Ito, Yoichi Nakao, Minoru Yoshida

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   489 ( 4 ) 455 - 459  2017.08  [Refereed]

     View Summary

    EGR2 is a zinc finger transcription factor that regulates myelination in the peripheral nervous system and T cell anergy. The transcriptional activity of EGR2 is known to be regulated by its co-activators and/or co-repressors. Although the activity of transcription factors is generally regulated not only by interactions with co-regulators but also posttranslational modifications including acetylation, little is known about posttranslational modifications of EGR2. Here we show that EGR2 is a novel acetylated protein. Through immunoblotting analyses using an antibody that specifically recognizes the acetylated form of EGR2, CBP and p300 were identified as acetyltransferases, while HDAC6, 10 and SIRT1 were identified as deacety-lases of EGR2. Although the NuRD complex containing HDAC1 and HDAC2 is known to associate with EGR2, the present study suggests that acetylation of EGR2 is regulated independently of NuRD. (C) 2017 Elsevier Inc. All rights reserved.

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  • A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B

    Shohei Takase, Rumi Kurokawa, Daisuke Arai, Kind Kanemoto Kanto, Tatsufumi Okino, Yoichi Nakao, Tetsuo Kushiro, Minoru Yoshida, Ken Matsumoto

    SCIENTIFIC REPORTS   7  2017.05  [Refereed]

     View Summary

    Genome-wide RNA interference (RNAi) with pooled and barcoded short-hairpin RNA (shRNA) libraries provides a powerful tool for identifying cellular components that are relevant to the modes/mechanisms of action (MoA) of bioactive compounds. shRNAs that affect cellular sensitivity to a given compound can be identified by deep sequencing of shRNA-specific barcodes. We used multiplex barcode sequencing technology by adding sample-specific index tags to PCR primers during sequence library preparation, enabling parallel analysis of multiple samples. An shRNA library screen with this system revealed that downregulation of ATP1A1, an a-subunit of Na+/K+ ATPase, conferred significant sensitivity to aurilide B, a natural marine product that induces mitochondria-mediated apoptosis. Combined treatment with ouabain which inhibits Na+/K+ ATPase by targeting a-subunits potentiated sensitivity to aurilide B, suggesting that ATP1A1 regulates mitochondria-mediated apoptosis. Our results indicate that multiplex sequencing facilitates the use of pooled shRNA library screening for the identification of combination drug therapy targets.

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  • In Vivo Gold Complex Catalysis within Live Mice

    Kazuki Tsubokura, Kenward K. H. Vong, Ambara R. Pradipta, Akihiro Ogura, Sayaka Urano, Tsuyoshi Tahara, Satoshi Nozaki, Hirotaka Onoe, Yoichi Nakao, Regina Sibgatullina, Almira Kurbangalieva, Yasuyoshi Watanabe, Katsunori Tanaka

    ANGEWANDTE CHEMIE-INTERNATIONAL EDITION   56 ( 13 ) 3579 - 3584  2017.03  [Refereed]

     View Summary

    Metal complex catalysis within biological systems is largely limited to cell and bacterial systems. In this work, a glycoalbumin-Au-III complex was designed and developed that enables organ-specific, localized propargyl ester amidation with nearby proteins within live mice. The targeted reactivity can be imaged through the use of Cy7.5- and TAMRA-linked propargyl ester based fluorescent probes. This targeting system could enable the exploitation of other metal catalysis strategies for biomedical and clinical applications.

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  • In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling

    Yamamizu Kohei, Iwasaki Mio, Takakubo Hitomi, Sakamoto Takumi, Ikuno Takeshi, Miyoshi Mami, Kondo Takayuki, Nakao Yoichi, Nakagawa Masato, Inoue Haruhisa, Yamashita Jun K.

    Stem Cell Reports   8  2017.02

     View Summary

    iPS細胞から血液脳関門モデルの作製に成功. 京都大学プレスリリース. 2017-02-27.The blood-brain barrier (BBB) is composed of four cell populations, brain endothelial cells (BECs), pericytes, neurons, and astrocytes. Its role is to precisely regulate the microenvironment of the brain through selective substance crossing. Here we generated an in vitro model of the BBB by differentiating human induced pluripotent stem cells (hiPSCs) into all four populations. When the four hiPSC-derived populations were co-cultured, endothelial cells (ECs) were endowed with features consistent with BECs, including a high expression of nutrient transporters (CAT3, MFSD2A) and efflux transporters (ABCA1, BCRP, PGP, MRP5), and strong barrier function based on tight junctions. Neuron-derived Dll1, which activates Notch signaling in ECs, was essential for the BEC specification. We performed in vitro BBB permeability tests and assessed ten clinical drugs by nanoLC-MS/MS, finding a good correlation with the BBB permeability reported in previous cases. This technology should be useful for research on human BBB physiology, pathology, and drug development.

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  • Trypanocidal activity, cytotoxicity and histone modifications induced by malformin A(1) isolated from the marine-derived fungus Aspergillus tubingensis IFM 63452

    K. Notarte, Y. Nakao, T. Yaguchi, M. Bungihan, K. Suganuma, T. E. dela Cruz

    MYCOSPHERE   8 ( 1 ) 111 - 120  2017  [Refereed]

     View Summary

    Malformin A(1), a cyclic pentapeptide, was isolated from the marine-derived fungus Aspergillus tubingensis IFM 63452. The identity of the compound was established based on TOFMS and H-1 NMR data. Malformin A(1) exhibited trypanocidal activity against Trypanosoma congolense (IC50: 15.08 ng/mL). Interestingly, the compound was selective for T. congolense rendering a selectivity index value that ranged from 3.33 to 4.67. It also demonstrated cytotoxicity against HeLa (IC50: 50.15 ng/mL) and P388 (IC50: 70.38 ng/mL) cell lines. To further identify the possible mechanism of its cytotoxic effect, immunofluorescence staining was conducted to follow the epigenetic changes induced by the compound in the amino acid lysine of histone H3 and H4 in HeLa. The compound induced repressive levels of H3K27me3, H3K27ac and H4K5ac, and enhanced levels of H3K9me2, H3K9me3 and H4K16ac supporting the compound's chemotherapeutic potential.

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  • One-Pot Evolution of AgeladineA through a Bio-Inspired Cascade towards Selective Modulators of Neuronal Differentiation

    Takayuki Iwata, Satoshi Otsuka, Kazuki Tsubokura, Almira Kurbangalieva, Daisuke Arai, Koichi Fukase, Yoichi Nakao, Katsunori Tanaka

    CHEMISTRY-A EUROPEAN JOURNAL   22 ( 41 ) 14707 - 14716  2016.10  [Refereed]

     View Summary

    A bio-inspired cascade reaction has been developed for the construction of the marine natural product ageladineA and a de novo array of its N1-substituted derivatives. This cascade features a 2-aminoimidazole formation that is modeled after an arginine post-translational modification and an aza-electrocyclization. It can be effectively carried out in a one-pot procedure from simple anilines or guanidines, leading to structural analogues of ageladineA that had been otherwise synthetically inaccessible. We found that some compounds out of this structurally novel library show a significant activity in modulating the neural differentiation. Namely, these compounds selectively activate or inhibit the differentiation of neural stem cells to neurons, while being negligible in the differentiation to astrocytes. This study represents a successful case in which the native biofunction of a natural product could be altered by structural modifications.

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  • Synthesis and antileishmanial activity of the core structure of cristaxenicin A

    Hitoshi Fumiyama, Tomoko Sadayuki, Yasutaka Osada, Yasuyuki Goto, Yoichi Nakao, Seijiro Hosokawa

    BIOORGANIC & MEDICINAL CHEMISTRY LETTERS   26 ( 17 ) 4355 - 217  2016.09  [Refereed]

     View Summary

    The core structure of cristaxenicin A having trans-fused dihydropyran and nine membered ring has been synthesized and evaluated the antileishmanial activity. The dihydropyran ring was synthesized by [4+2] cycloaddition reaction between an unsaturated aldehyde and a beta-alkoxy-alpha,beta-unsaturated ketone. The nine membered ring possessing alpha,beta-unsaturated aldehyde was constructed by the intramolecular NHK reaction followed by the Mitsunobu rearrangement. The racemic core structure of cristaxenicin A was evaluated the anti-leishmanial activity with an IC50 value of 2.4 mu M. (C) 2016 Elsevier Ltd. All rights reserved.

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    5
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  • A Genetically Encoded FRET Probe to Detect Intranucleosomal Histone H3K9 or H3K14 Acetylation Using BRD4, a BET Family Member

    Shiho Nakaoka, Kazuki Sasaki, Akihiro Ito, Yoichi Nakao, Minoru Yoshida

    ACS CHEMICAL BIOLOGY   11 ( 3 ) 729 - 733  2016.03  [Refereed]

     View Summary

    Acetylation is a well-characterized histone modification, which plays important roles in controlling epigenetic gene expression, and its malfunction is tightly associated with cancer. By taking advantage of the specific binding of BRD4 to acetylated lysine residues, we developed a FRET-based probe for visualizing histone H3 acetylation in living cells. BRD4, a protein known to be involved in acute myeloid leukemia and nuclear protein in testis midline carcinoma, recognizes the acetylation of histone H3 via its bromodomains. The probe exhibited a significant change in FRET signaling that was dependent on histone H3 acetylation. Mutagenesis studies revealed that an increase in the emission ratio reflected the acetylation of either K9 or K14 of histone H3 within the probe. Since BRD4 has increasingly drawn attention as a new anticancer drug target, we demonstrated that the developed fluorescent probe will also serve as a powerful tool to evaluate BRD4 inhibitors in living cells.

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  • Piceatannol is superior to resveratrol in promoting neural stem cell differentiation into astrocytes

    Daisuke Arai, Ryousuke Kataoka, Satoshi Otsuka, Midori Kawamura, Hiroko Maruki-Uchida, Masahiko Sai, Tatsuhiko Ito, Yoichi Nakao

    FOOD & FUNCTION   7 ( 10 ) 4432 - 4441  2016  [Refereed]

     View Summary

    Piceatannol (3,3', 4', 5-trans-tetrahydroxystilbene) is a polyphenolic compound abundant in the seeds of passion fruit (Passiflora edulis). Piceatannol is an analogue of resveratrol (3, 4', 5-trans-trihydroxystilbene) and shares the structural motif and biological activities such as activation of SIRT1. Several studies have shown that piceatannol is more potent than resveratrol. In this study, we examined the effects of piceatannol on neural stem cell differentiation into astrocytes compared with those of resveratrol. At a concentration of 2.5 mu M, piceatannol promoted astrocyte differentiation, while resveratrol had no effect at this concentration. Furthermore, we found that oral administration of piceatannol increased the number of astrocytes in the brains of adult mice, while resveratrol administration showed no effects. These results suggest that piceatannol has a superior effect to resveratrol in promoting astrocyte differentiation.

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  • An epigenetic regulatory element of the Nodal gene in the mouse and human genomes

    Daisuke Arai, Koji Hayakawa, Jun Ohgane, Mitsuko Hirosawa, Yoichi Nakao, Satoshi Tanaka, Kunio Shiota

    MECHANISMS OF DEVELOPMENT   136   143 - 154  2015.05  [Refereed]

     View Summary

    Nodal signaling plays critical roles during embryonic development. The Nodal gene is not expressed in adult tissues but is frequently activated in cancer cells, contributing to progression toward malignancy. Although several regulatory elements of the Nodal gene have been identified, the epigenetic mechanisms by which Nodal expression is regulated over the long term remain unclear. We found a region exhibiting dynamic changes in DNA methylation at approximately -3.0 kb to -0.4 kb upstream from the transcriptional start site (TSS) that we termed the epigenetic regulatory element (ERE). The ERE was unmethylated in mouse embryonic stem cells (mESCs) but became increasingly methylated in differentiated cells and tissues, concomitant with the downregulation of Nodal mRNA expression. In vitro reporter assays identified an Oct3/4 binding motif within the ERE, indicating that the ERE is responsible for the activation of Nodal in mESCs. Furthermore, the ERE was a target of differentiation-associated Polycomb silencing, and the chromatin condensed when mESCs differentiated to embryoid bodies (EBs). Pharmacological inhibition of PRC2 led to the reactivation of Nodal expression in EBs and mouse embryonic fibroblasts (MEFs). The ERE was also targeted by PRC2 in normal human cells. In NODAL-expressing human cancer cells, accumulation of EZH2 and trimethylation of H3K27 at the ERE were diminished. In conclusion, Nodal is epigenetically controlled through the ERE in the mouse embryo and human cells. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

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  • Direct Guanylation of Amino Groups by Cyanamide in Water: Catalytic Generation and Activation of Unsubstituted Carbodiimide by Scandium(III) Triflate

    Kazuki Tsubokura, Takayuki Iwata, Misako Taichi, Almira Kurbangalieva, Koichi Fukase, Yoichi Nakao, Katsunori Tanaka

    SYNLETT   25 ( 9 ) 1302 - 1306  2014.06  [Refereed]

     View Summary

    Guanylation proceeded efficiently upon treatment of the various amines with cyanamide in the presence of catalytic amounts of scandium(III) triflate under mild conditions. The method did not require the guanylation reagents to be preactivated, and the reaction proceeded efficiently in water. The method, therefore, has practical utility for substrates that dissolve only in aqueous solutions, for example, peptides or pharmacologically important compounds.

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    23
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  • Inhibition of protein SUMOylation by davidiin, an ellagitannin from Davidia involucrata

    Misao Takemoto, Yumi Kawamura, Mikako Hirohama, Yuki Yamaguchi, Hiroshi Handa, Hisato Saitoh, Yoichi Nakao, Manabu Kawada, Khan Khalid, Hiroyuki Koshino, Ken-ichi Kimura, Akihiro Ito, Minoru Yoshida

    JOURNAL OF ANTIBIOTICS   67 ( 4 ) 335 - 338  2014.04  [Refereed]

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  • Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system

    Mikako Hirohama, Arnout R. D. Voet, Takeaki Ozawa, Hisato Saitoh, Yoichi Nakao, Kam Y. J. Zhang, Akihiro Ito, Minoru Yoshida

    ANALYTICAL BIOCHEMISTRY   448   92 - 94  2014.03  [Refereed]

     View Summary

    SUMOylation is a posttranslational process that attaches a small ubiquitin-like modifier (SUMO) to its target proteins covalently. SUMOylation controls multiple cellular processes through the recognition of SUMO by a SUMO-interacting motif (SIM). In this study, we developed assay systems for detecting noncovalent interactions between SUMO and SIM in cells using split-luciferase complementation. We applied a version of this assay to the detection of in vitro SUMO-SIM interactions using a bacterial expression system. These novel assays enable screening of inhibitors of SUMO-dependent protein-protein interactions, either in vivo or in vitro, in a high-throughput manner. (C) 2013 Elsevier Inc. All rights reserved.

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  • Cinanthrenol A, an Estrogenic Steroid Containing Phenanthrene Nucleus, from a Marine Sponge Cinachyrella sp.

    Koshi Machida, Takahiro Abe, Daisuke Arai, Mayumi Okamoto, Isao Shimizu, Nicole J. de Voogd, Nobuhiro Fusetani, Yoichi Nakao

    ORGANIC LETTERS   16 ( 6 ) 1539 - 1541  2014.03  [Refereed]

     View Summary

    Cinanthrenol A (1), a new steroid composed of a phenanthrene and a spiro[2,4]heptane system, was isolated from the marine sponge Cinachyrella sp. It is the first phenathrene-containing steroid with estrogen activity.

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  • Insights on pregnane-X-receptor modulation. Natural and semisynthetic steroids from Theonella marine sponges

    Valentina Sepe, Claudio D'Amore, Raffella Ummarino, Barbara Renga, Maria Valeria D'Auria, Ettore Novellino, Annamaria Sinisi, Orazio Taglialatela-Scafati, Yoichi Nakao, Vittorio Limongelli, Angela Zampella, Stefano Fiorucci

    EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY   73   126 - 134  2014.02  [Refereed]

     View Summary

    Pregnane-X-receptor (PXR) is a member of nuclear receptors superfamily that activates gene transcription by binding to responsive elements in the promoter of target genes. PXR is a master gene orchestrating the expression/activity of genes involved in the metabolism of endobiotics including bilirubin, bile acids, glucose and lipid. In addition PXR oversights the metabolism of the large majority of xenobiotics including a large amount of prescribing drugs. Thus, developing PXR ligands represents a great opportunity for a therapeutic intervention on human diseases including diabetes, obesity, dyslipidemias and liver disorders. To this end, natural compounds represent an arsenal of new chemical scaffolds useful for the identification of novel PXR ligands. Here, we report a series of 4-methylenesteroid derivatives isolated from Theonella marine sponges as novel PXR modulators. In addition, combining medicinal chemistry, pharmacological experiments and computational studies, we have investigated the effects of different modifications on ring A and on the side chain of 4-methylenesteroid derivatives toward PXR modulation. This study provides the molecular bases of ligand/PXR interaction useful for designing novel PXR modulators. (C) 2013 Elsevier Masson SAS. All rights reserved.

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  • 11C-Labeling of the C(1)-C(10) dihydroxy acid moiety for the study on the synthesis of kulokekahilide-2 PET tracer.

    Han, C.G, Doi, H, Kimura, J, Nakao, Y, Suzuki, M

    Int. J. Org. Chem.   4   269 - 277  2014  [Refereed]

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  • Spectomycin B1 as a novel sumoylation inhibitor that directly binds to SUMO E2

    Mikako Hirohama, Ashutosh Kumar, Isao Fukuda, Seiji Matsuoka, Yasuhiro Igarashi, Hisato Saitoh, Motoki Takagi, Kazuo Shin-Ya, Kaori Honda, Yasumitsu Kondoh, Tamio Saito, Yoichi Nakao, Hiroyuki Osada, Kam Y. J. Zhang, Minoru Yoshida, Akihiro Ito

    ACS Chemical Biology   8 ( 12 ) 2635 - 2642  2013.12  [Refereed]

     View Summary

    Conjugation of small ubiquitin-like modifier (SUMO) to protein (SUMOylation) regulates multiple biological systems by changing the functions and fates of a large number of proteins. Consequently, abnormalities in SUMOylation have been linked to multiple diseases, including breast cancer. Using an in situ cell-based screening system, we have identified spectomycin B1 and related natural products as novel SUMOylation inhibitors. Unlike known SUMOylation inhibitors such as ginkgolic acid, spectomycin B1 directly binds to E2 (Ubc9) and selectively blocks the formation of the E2-SUMO intermediate
    that is, Ubc9 is the direct target of spectomycin B1. Importantly, either spectomycin B1 treatment or Ubc9 knockdown inhibited estrogen-dependent proliferation of MCF7 human breast-cancer cells. Our findings suggest that Ubc9 inhibitors such as spectomycin B1 have potential as therapeutic agents against hormone-dependent breast cancers. © 2013 American Chemical Society.

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  • Stereochemical analysis and cytotoxicity of kulokekahilide-2 and its analogues

    Masahiro Umehara, Takayuki Negishi, Yukie Maehara, Yoichi Nakao, Junji Kimura

    TETRAHEDRON   69 ( 14 ) 3045 - 3053  2013.04  [Refereed]

     View Summary

    We report the conformational analysis of kulokekahilide-2, a cytotoxic cyclic depsipeptide, and its analogues. We also evaluated their cytotoxicity against human cancer cells. Although both the cis and trans conformations are possible for the amide bond between MePhe and MeGly, only one conformation was observed in DMSO. We also reveal that the configuration at C-43 in kulokekahilide-2 controls intramolecular ester exchange between the 26- and 24-membered cyclic depsipeptides. Kulokekahilide-2 and its analogues were evaluated for their cell-growth inhibition profile and using COMPARE analysis, which suggested a mechanism of action different from that of standard anticancer drugs. (C) 2013 Elsevier Ltd. All rights reserved.

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  • Structure-related cytotoxic activity of derivatives from kulokekahilide-2, a cyclodepsipeptide in Hawaiian marine mollusk

    Masahiro Umehara, Takayuki Negishi, Tomoko Tashiro, Yoichi Nakao, Junji Kimura

    BIOORGANIC & MEDICINAL CHEMISTRY LETTERS   22 ( 24 ) 7422 - 7425  2012.12  [Refereed]

     View Summary

    Kulokekahilide-2, a 26-membered cyclodepsipeptide, was isolated from Hawaiian marine mollusk and possessed potent cytotoxicity in mammalian tumor cells. In the present study, we synthesized kulokekahilide-2 and its derivatives and examined the structure-activity relationships of these peptides in human cancer cells (A549, K562, and MCF7 cells). This study demonstrated that the cyclization of depsipeptide and the chirality of the 21 position in Ala in kulokekahilide-2 were important for its cytotoxic property and that addition of halogen at the para position of phenyl group in the 24-D-MePhe in kulokekahilide-2 as well as some derivatives remarkably increased their cytotoxicity in human cancer cells. These results suggest that the modifications of 24-D-MePhe in kulokekahilide-2, preserving its cyclization and the chirality at the 21-position, are promising strategy for exploring new derivative of kulokekahilide-2 as anti-tumor drug. (c) 2012 Elsevier Ltd. All rights reserved.

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  • Cristaxenicin A, an Antiprotozoal Xenicane Diterpenoid from the Deep Sea Gorgonian Acanthoprimnoa cristata

    Shin-Taro Ishigami, Yasuyuki Goto, Noboru Inoue, Shin-Ichiro Kawazu, Yoshitsugu Matsumoto, Yukimitsu Imahara, Moto Tarumi, Hiromi Nakai, Nobuhiro Fusetani, Yoichi Nakao

    JOURNAL OF ORGANIC CHEMISTRY   77 ( 23 ) 10962 - 10966  2012.12  [Refereed]

     View Summary

    A new xenicane diterpenoid, cristaxenicin A (1), has been isolated from the deep sea gorgonian Acanthoprimnoa cristata. The structure of 1 was elucidated on the basis of spectral analysis including NMR and MS. The absolute configuration of 1 was determined on the basis of quantum chemical calculation of CD spectra. Cristaxenicin A (1) showed antiprotozoal activities against Leishmania amazonensis and Trypanosoma congolense with IC50 values of 0.088 and 0.25 mu M, respectively.

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  • Preliminary Structure-Activity Relationship on Theonellasterol, a New Chemotype of FXR Antagonist, from the Marine Sponge Theonella swinhoei

    Valentina Sepe, Raffaella Ummarino, Maria Valeria D'Auria, Orazio Taglialatela-Scafati, Simona De Marino, Claudio D'Amore, Barbara Renga, Maria Giovanna Chini, Giuseppe Bifulco, Yoichi Nakao, Nobuhiro Fusetani, Stefano Fiorucci, Angela Zampella

    MARINE DRUGS   10 ( 11 ) 2448 - 2466  2012.11  [Refereed]

     View Summary

    Using theonellasterol as a novel FXR antagonist hit, we prepared a series of semi-synthetic derivatives in order to gain insight into the structural requirements for exhibiting antagonistic activity. These derivatives are characterized by modification at the exocyclic carbon-carbon double bond at C-4 and at the hydroxyl group at C-3 and were prepared from theonellasterol using simple reactions. Pharmacological investigation showed that the introduction of a hydroxyl group at C-4 as well as the oxidation at C-3 with or without concomitant modification at the exomethylene functionality preserve the ability of theonellasterol to inhibit FXR transactivation caused by CDCA. Docking analysis showed that the placement of these molecules in the FXR-LBD is well stabilized when on ring A functional groups, able to form hydrogen bonds and pi interactions, are present.

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  • The total synthesis and structure-activity relationships of a highly cytotoxic depsipeptide kulokekahilide-2 and its analogs

    Yuuki Takada, Masahiro Umehara, Ryosuke Katsumata, Yoichi Nakao, Junji Kimura

    TETRAHEDRON   68 ( 2 ) 659 - 669  2012.01  [Refereed]

     View Summary

    We successfully completed the total synthesis of kulokekahilide-2, a cytotoxic depsipeptide from the Cephalaspiedean mollusk Phillinopsis speciosa. We have revised the absolute stereochemistry of kulokekahilide-2 to 21-L-Ala, 24-D-MePhe, 37-L-Ile, 43-D-Ala, 15-D-Hica, and 5S,6S,7S-Dtda. We also investigated the cause of the mis-assignment of the configuration in the originally proposed kulokekahilide-2 and concluded that methanolysis using MeONa caused partial racemization, which led to the mis-assignment. The structure activity relationships of kulokekahilide-2 and its analogs indicate the importance of an L-amino acid at position 21. (C) 2011 Elsevier Ltd. All rights reserved.

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    16
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  • Shishicrellastatins, inhibitors of cathepsin B, from the marine sponge Crella (Yvesia) spinulata

    Shuhei Murayama, Yasufumi Imae, Kentaro Takada, Jo Kikuchi, Yoichi Nakao, Rob W. M. van Soest, Shigeru Okada, Shigeki Matsunaga

    BIOORGANIC & MEDICINAL CHEMISTRY   19 ( 22 ) 6594 - 6598  2011.11  [Refereed]

     View Summary

    Two dimeric steroid derivatives, shishicrellastatin A (1) and B (2), have been isolated as cathepsin B inhibitors from the marine sponge Crella (Yvesia) spinulata. Their structures were determined by interpretation of spectroscopic data. Shishicrellastatins inhibit cathepsin B with an IC(50) value of 8 mu g/mL each. (C) 2011 Elsevier Ltd. All rights reserved.

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  • Halenaquinone, a chemical compound that specifically inhibits the secondary DNA binding of RAD51

    Motoki Takaku, Takashi Kainuma, Takako Ishida-Takaku, Shintaro Ishigami, Hidekazu Suzuki, Satoshi Tashiro, Rob W. M. van Soest, Yoichi Nakao, Hitoshi Kurumizaka

    GENES TO CELLS   16 ( 4 ) 427 - 436  2011.04  [Refereed]

     View Summary

    Mutations and single-nucleotide polymorphisms affecting RAD51 gene function have been identified in several tumors, suggesting that the inappropriate expression of RAD51 activity may cause tumorigenesis. RAD51 is an essential enzyme for the homologous recombinational repair (HRR) of DNA double-strand breaks. In the HRR pathway, RAD51 catalyzes the homologous pairing between single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), which is the central step of the HRR pathway. To identify a chemical compound that regulates the homologous-pairing activity of RAD51, in the present study, we screened crude extract fractions from marine sponges by the RAD51-mediated homologous-pairing assay. Halenaquinone was identified as an inhibitor of the RAD51 homologous-pairing activity. A surface plasmon resonance analysis indicated that halenaquinone directly bound to RAD51. Intriguingly, halenaquinone specifically inhibited dsDNA binding by RAD51 alone or the RAD51-ssDNA complex, but only weakly affected the RAD51-ssDNA binding. In vivo, halenaquinone significantly inhibited the retention of RAD51 at double-strand break sites. Therefore, halenaquinone is a novel type of RAD51 inhibitor that specifically inhibits the RAD51-dsDNA binding.

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    37
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  • A One-Pot Synthesis and Biological Activity of Ageladine A and Analogues

    Sudhir R. Shengule, Wendy L. Loa-Kum-Cheung, Christopher R. Parish, Melina Blairvacq, Laurent Meijer, Yoichi Nakao, Peter Karuso

    JOURNAL OF MEDICINAL CHEMISTRY   54 ( 7 ) 2492 - 2503  2011.04  [Refereed]

     View Summary

    A one-pot synthesis of ageladine A and analogues is reported. The key Pictet - Spengler reaction between 2-aminohistamine and aryl aldehydes has been successfully utilized for the synthesis of the natural product and 14 analogues. These compounds were screened for their matrix metalloprotease (MMP) and kinase inhibition to develop the first structure-activity relationship of ageladine A analogues. One compound, which showed significant kinase activity but little MMP inhibitory activity, was found to be highly active in an antiangiogenic screen, suggesting that the angiogenic activity of ageladine A is not associated with MMP inhibition but rather kinase inhibitory activity. Cytotoxicity was excluded as a mode of action by the assay of ageladine A and an analogue against 60 human cell lines.

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  • Marine invertebrates: Sponges

    Yoichi Nakao, Nobuhiro Fusetani

    Comprehensive Natural Products II: Chemistry and Biology   2   327 - 362  2010.03

  • Gracilioethers A-C, Antimalarial Metabolites from the Marine Sponge Agelas gracilis

    Reiko Ueoka, Yoichi Nakao, Shizuka Kawatsu, Junko Yaegashi, Yoshitsugu Matsumoto, Shigeki Matsunaga, Kazuo Furihata, Rob W. M. van Soest, Nobuhiro Fusetani

    JOURNAL OF ORGANIC CHEMISTRY   74 ( 11 ) 4203 - 4207  2009.06  [Refereed]

     View Summary

    Three new antiprotozoan compounds, gracilioethers A-C (1-3), have been isolated from the marine sponge Agelas gracilis. Their structures were elucidated on the basis of spectroscopic and chemical methods. Gracilioethers A-C showed antimalarial activity against Plasmodium falciparum with IC(50) values of 0.5-10 mu g/mL, whereas gracilioether B (2) also showed antileishmanial activity.

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    69
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  • A new antiangiogenic C(24) oxylipin from the soft coral Sinularia numerosa

    Takahiro Yamashita, Yoichi Nakao, Shigeki Matsunaga, Tsutomu Oikawa, Yukimitsu Imahara, Nobuhiro Fusetani

    BIOORGANIC & MEDICINAL CHEMISTRY   17 ( 6 ) 2181 - 2184  2009.03  [Refereed]

     View Summary

    A new oxylipin, 15-hydroxy-tetracosa-6,9,12,16,18-pentaenoic acid (15-HTPE; 1) was isolated as an inhibitor of tube-formation from the soft coral Sinularia numerosa. Its structure was elucidated by means of spectral analysis and chemical degradation. 15-HTPE inhibited tube formation of EA.hy926 cells at the concentration of 20-40 mu M. (C) 2008 Elsevier Ltd. All rights reserved.

    DOI

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    14
    Citation
    (Scopus)
  • Intramolecular ester exchange of potent cytotoxic kulokekahilide-2

    Masahiro Umehara, Yuuki Takada, Yoichi Nakao, Junji Kimura

    TETRAHEDRON LETTERS   50 ( 7 ) 840 - 843  2009.02  [Refereed]

     View Summary

    Kulokekahilide-2 is a 26-membered cyclic depsipeptide that exhibits potent cytotoxicity against HeLa and P388 cells with IC(50) values of 3.2 and 16 ng/mL, respectively. We have achieved a total synthesis of natural kulokekahilide-2, the NMR spectra of which showed complex signals because of trans and cis conformers at the amide bond. Besides, the spectra revealed that a mixture of 26- and 24-membered cyclic depsipeptides had been produced due to intramolecular ester exchange. The isolated 24-membered compound was transformed into the 26-membered compound over a period of several days. The two isomers have been shown to be in equilibrium and to display almost the same cytotoxicity. (C) 2008 Elsevier Ltd. All rights reserved.

    DOI

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    10
    Citation
    (Scopus)
  • Koshikamide B, a cytotoxic peptide lactone from a marine sponge Theonella sp

    Takahiro Araki, Shigeki Matsunaga, Yoichi Nakao, Kazuo Furihata, Lyndon West, D. John Faulkner, Nobuhiro Fusetani

    JOURNAL OF ORGANIC CHEMISTRY   73 ( 20 ) 7889 - 7894  2008.10  [Refereed]

     View Summary

    Koshikamide B (1) has been isolated from two separate collections of the marine sponge Theonella sp. as the major cytotoxic constituent. Koshikamide B is a 17-residue peptide lactone composed of six proteinogenic amino acids, two D-isomers of proteinogenic amino acids, seven N-methylated amino acids, and two unusual amino acid residues. The unusual amino acids are N(delta)-carbamoylasparagine and 2-(3-amino-2-hydroxy-5-oxopyrrolidin-2-yl)propionic acid (AHPP); the former is first found as the constituent of peptides, whereas the latter is a new amino acid residue. The N-terminus of koshikamide B is blocked by a methoxyacetyl group. The structure of koshikamide B (1) has been determined by interpretation of spectral data and analysis of chemical degradation products. Koshikamide B (1) exhibits cytotoxicity against P388 murine leukemia cells and the human colon tumor (HCT-116) cell line with an IC(50) value of 0.45 and 7.5 mu g/mL, respectively.

    DOI

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    28
    Citation
    (Scopus)
  • Aplysinoplides A-C, cytotoxic sesterterpenes from the marine sponge Aplysinopsis digitata

    Reiko Ueoka, Yoichi Nakao, Shinobu Fujii, Rob W. M. van Soest, Shigeki Matsunaga

    JOURNAL OF NATURAL PRODUCTS   71 ( 6 ) 1089 - 1091  2008.06  [Refereed]

     View Summary

    Three sesterterpenoids, aplysinoplides A -C (1-3), were isolated from the marine sponge Aplysinopsis digitata. Their structures were determined on the basis of spectroscopic data. They exhibited cytotoxic activity against P388 mouse leukemia cells.

    DOI

    Scopus

    24
    Citation
    (Scopus)
  • Asteropterin, an inhibitor of cathepsin B, from the marine sponge Asteropus simplex

    Shuhei Murayama, Yoichi Nakao, Shigeki Matsunaga

    TETRAHEDRON LETTERS   49 ( 26 ) 4186 - 4188  2008.06  [Refereed]

     View Summary

    A new pteridine derivative, asteropterin (1), was isolated as a cathepsin B inhibitor from the marine sponge Asteropus simplex. The structure of asteropterin (1) was elucidated by the analysis of spectral data. (C) 2008 Elsevier Ltd. All rights reserved.

    DOI

    Scopus

    15
    Citation
    (Scopus)
  • Evaluation of antiangiogenic activity of azumamides by the in vitro vascular organization model using mouse induced pluripotent stem (iPS) cells

    Yoichi Nakao, Genta Narazaki, Takuhiro Hoshino, Satoko Maeda, Minoru Yoshida, Hiroshi Maejima, Jun K. Yamashita

    BIOORGANIC & MEDICINAL CHEMISTRY LETTERS   18 ( 9 ) 2982 - 2984  2008.05  [Refereed]

     View Summary

    Evaluation of antiangiogenic activity of marine sponge derived azumamides by the in vitro vascular organization model using mouse induced pluripotent stem (iPS) cells was carried out. Azumamide E (5) strongly inhibited in vitro angiogenesis from iPS cells at 1.9 mu M while azumamide A (1) showed only weak inhibition at 19 mu M. These results were well correlated with HDAC inhibitory activity of these compounds, revealing the prospect of azumamides as the probe molecules useful for stem cell chemical biology. (C) 2008 Published by Elsevier Ltd.

    DOI

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    27
    Citation
    (Scopus)
  • Ciliatamides A-C, bioactive lipopeptides from the deep-sea sponge Aaptos ciliatao

    Yoichi Nakao, Shizuka Kawatsu, Chikane Okamoto, Masaaki Okamoto, Yoshitsugu Matsumoto, Shigeki Matsunaga, Rob. W. M. van Soest, Nobuhiro Fusetani

    JOURNAL OF NATURAL PRODUCTS   71 ( 3 ) 469 - 472  2008.03  [Refereed]

     View Summary

    Three lipopeptides, ciliatamides A-C (1-3), were isolated from the deep-sea sponge Aaptos ciliata, and their structures were elucidated on the basis of spectroscopic and chemical methods. Ciliatamides A (1) and B (2) were found to be antileishmanial, while 2 also exhibited marginal cytotoxicity to HeLa cells.

    DOI

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    32
    Citation
    (Scopus)
  • Revised absolute stereochemistry of natural kulokekahilide-2

    Yuuki Takada, Masahiro Umehara, Yoichi Nakao, Junji Kimura

    TETRAHEDRON LETTERS   49 ( 7 ) 1163 - 1165  2008.02  [Refereed]

     View Summary

    Kulokekahilide-2 is a potent cytotoxic depsipeptide isolated from the Hawaiian marine mollusk Philinopsis speciosa. The structure of kutokekahilide-2 was reported to be composed of five amino acids (L-Ala, L-Ile, MeGly, L-MePhe, D-Ala) and two hydroxy acids (D-Hica, 5S,6S,7S-Dtda); however, following its total synthesis, the (1)H NMR spectrum of the synthetic compound was found to be different from that of the natural one, suggesting that the stereochemistry of the reported structure was incorrect. To determine the stereochemistry of the natural compound, arrays of analogues have been prepared using different sets of chiral amino acids, and the absolute stereochemistry of kulokekahilide-2 has been unambiguously confirmed to involve the combination 21-L-Ala, 24-D-MePhe, and 43-D-Ala. (C) 2007 Elsevier Ltd. All rights reserved.

    DOI

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    19
    Citation
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  • Cytotoxic C-47-polyacetylene carboxylic acids from a marine Sponge pertrosia sp.

    Chikane Okamoto, Yoichi Nakao, Tsuyoshi Fujita, Takashi Iwashita, W. M. van Soest, Nobuhiro Fusetani, Shigeki Matsunaga

    JOURNAL OF NATURAL PRODUCTS   70 ( 11 ) 1816 - 1819  2007.11  [Refereed]

     View Summary

    Two new polyacetylene carboxylic acids, petroformynic acids B (3) and C (4), were isolated from a marine sponge Pertrosia sp. as cytotoxic constituents. Their structures were determined by interpretation of 2D NMR data and tandem FABMS data. Absolute stereochemistry of 3 was assigned by application of the modified Mosher analysis. Petroformynic acids exhibit moderate cytotoxic activity against P388 cells.

    DOI

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    18
    Citation
    (Scopus)
  • Absolute structure of prunustatin A, a novel GRP78 molecular chaperone down-regulator

    Yukiko Umeda, Kazuo Furihata, Shohei Sakuda, Hiromichi Nagasawa, Ken Ishigami, Hidenori Watanabe, Who Izumikawa, Motoki Takagi, Takayuki Doi, Yoichi Nakao, Kazuo Shin-ya

    ORGANIC LETTERS   9 ( 21 ) 4239 - 4242  2007.10  [Refereed]

     View Summary

    In the course of our screening program for regulators of a molecular chaperone GRP78 expression, we isolated a novel inhibitor of GRP78 expression, designated as prunustatin A, from Streptomyces violaceoniger 4521-SVS3. The planar structure of prunustatin A was determined to be an oxidized type of the neoantimycin family. Its absolute stereochemistry was established to be 2R, 4S, 6S, 7R, 9S, and 29S by analyzing chemically degraded components obtained from the derivative of prunustatin A.

    DOI

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    27
    Citation
    (Scopus)
  • The structures of three new shishididemniols from a tunicate of the family Didemnidae

    Hirotsugu Kobayashi, Yoshinari Miyata, Kohtaro Okada, Tsuyoshi Fujita, Takashi Iwashita, Yoichi Nakao, Nobuhiro Fusetani, Shigeki Matsunaga

    TETRAHEDRON   63 ( 29 ) 6748 - 6754  2007.07  [Refereed]

     View Summary

    Three new serinolipid derivatives, shishididemniols C (1), D (2), and E (3), were isolated as antibacterial constituents of a tunicate of the family Didemnidae. Their planar structures were elucidated by interpretation of NMR and MS data, whereas the absolute stereochemistry was determined by chemical conversions. Shishididemniols C (3), D (4), and E (5) exhibited antibacterial activity against the fish pathogenic bacterium Vibrio anguillarum. similar to 2007 Published by Elsevier Ltd.

    DOI

    Scopus

    12
    Citation
    (Scopus)
  • Application of a 6 pi-1-azatriene electrocyclization strategy to the total synthesis of the marine sponge metabolite ageladine A and biological evaluation of synthetic analogues

    Matthew L. Meketa, Steven M. Weinreb, Yoichi Nakao, Nobuhiro Fusetani

    JOURNAL OF ORGANIC CHEMISTRY   72 ( 13 ) 4892 - 4899  2007.06  [Refereed]

     View Summary

    A 12-step synthesis of the angiogenesis inhibitory marine metabolite ageladine A is reported. The key steps include a 6 pi-1-azatriene electrocyclization for formation of the pyridine ring and a Suzuki-Miyaura coupling of N-Boc-pyrrole-2-boronic acid with a chloroimidazopyridine. In addition, an assessment of the biological activity of a variety of synthetic analogues of ageladine A prepared during this synthesis is described.

    DOI

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    42
    Citation
    (Scopus)
  • Carteramine A, an inhibitor of neutrophil chemotaxis, from the marine sponge Stylissa carteri

    Hirotsugu Kobayashi, Koki Kitamura, Koji Nagai, Yoichi Nakao, Nobuhiro Fusetani, Rob W. M. van Soest, Shigeki Matsunaga

    TETRAHEDRON LETTERS   48 ( 12 ) 2127 - 2129  2007.03  [Refereed]

     View Summary

    A new dimeric oroidin derivative, carteramine A (1), was isolated as a neutrophil chemotaxis inhibitor from the marine sponge Stylissa carteri. The structure of 1 was elucidated by the analysis of spectral data. (c) 2007 Elsevier Ltd. All rights reserved.

    DOI

    Scopus

    89
    Citation
    (Scopus)
  • Total synthesis of azumamide A and azumamide E, evaluation as histone deacetylase inhibitors, and design of a more potent analogue

    Shijun Wen, Krystle L. Carey, Yoichi Nakao, Nobuhiro Fusetani, Graham Packham, A. Ganesan

    ORGANIC LETTERS   9 ( 6 ) 1105 - 1108  2007.03  [Refereed]

     View Summary

    The unprecedented diastereoselective Mannich reaction of a Z-allylsulfoximine was a key step in the total synthesis of the marine natural products azumamide A and E, and an unnatural analogue. Their relative potency as histone deacetylase inhibitors was evaluated and found to correlate with predicted zinc-binding affinity.

    DOI

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    55
    Citation
    (Scopus)
  • Complete structure elucidation of shishididemniols, complex lipids with tyramine-derived tether and two serinol units, from a marine tunicate of the family didemnidae

    Hirotsugu Kobayashi, Jun'ichiro Ohashi, Tsuyoshi Fujita, Takashi Iwashita, Yoichi Nakao, Shigeki Matsunaga, Nobuhiro Fusetani

    JOURNAL OF ORGANIC CHEMISTRY   72 ( 4 ) 1218 - 1225  2007.02  [Refereed]

     View Summary

    Two new serinolipid derivatives, shishididemniols A (1) and B (2), were isolated as antibacterial constituents of a tunicate of the family Didemnidae. The structure of 1 was elucidated by interpretation of spectral data and the application of the modified Mosher method to 1 and its suitable degradation products. Compound 2 was the chlorohydrin of 1. Compounds 1 and 2 exhibited antibacterial activity against fish pathogenic bacterium Vibrio anguillarum.

    DOI

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    22
    Citation
    (Scopus)
  • Reinvestigation of the stereochemistry of kulokekahilide-2

    Takada, Y, Mori, E, Umehara, M, Nakao, Y, Kimura, J

    Tetrahedron Lett.   48   4892 - 4899  2007  [Refereed]

  • Asteropine A, a sialidase-inhibiting conotoxin-like peptide from the marine sponge Asteropus simplex

    Takada, K, Hamada,T, Hirota, H, Nakao, Y, Matsunaga, S, van Soest, R. W. M, Fusetani, N

    Chem. Biol.   13   569 - 574  2006  [Refereed]

  • Azumamides A-E, new HDAC inhibitory cyclic tetrapeptides from the marine sponge Mycale izuensis

    Nakao, Y, Yoshida, S, Matsunaga, S, Shindoh, N, Terada, Y, Nagai, K, Yamashita, J. K, Ganesan, A, van Soest, R. W. M, Fusetani N

    Angew. Chem. Int. Ed.   45   7553 - 7557  2006  [Refereed]

    Authorship:Lead author

  • Structure-activity relationship study on 13-deoxytedanolide, a highly antitumor macrolide from the marine sponge Mycale adhaerens

    Nishimura, S, Matsunaga, S, Yoshida, S, Nakao, Y, Hirota, H, Fusetani, N

    Bioorg. Med. Chem.   13   455 - 462  2005  [Refereed]

    DOI CiNii

  • Axinelloside A, an unprecedented highly sulfated lipopolysaccharide inhibiting telomerase, from the marine sponge, Axinella infundibula

    Warabi, K, Hamada, T, Nakao, Y, Matsunaga, S, Hirota, H, van Soest, R. W. M, Fusetani, N

    J. Am. Chem. Soc.   127   13262 - 13270  2005  [Refereed]

  • Penasulfate A, a new -glucosidase inhibitor from a marine sponge Penares sp.

    Nakao, Y, Maki, T, Matsunaga, S, Fusetani, N

    J. Nat. Prod.   67   1346 - 1350  2004  [Refereed]

    Authorship:Lead author

  • Kulokekahilide-2, a cytotoxic depsipeptide from a cephalaspidean mollusk Philinopsis speciosa

    Nakao, Y, Yoshida, W. Y, Takada, Y, Kimura, J, Yang, L, Mooberry, S. L, Scheuer, P. J

    J. Nat. Prod.   67   1332 - 1340  2004  [Refereed]

    Authorship:Lead author

  • Identification of Renieramycin A as an antileishmanial substance in a marine sponge Neopetrosia sp

    Nakao Y, Shiroiwa T, Murayama S, Matsunaga S, Goto Y, Matsumoto Y, Fusetani N

    Marine Drugs   2   55 - 62  2004  [Refereed]

    DOI

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    49
    Citation
    (Scopus)
  • Schulzeine A-C, New Glycosidase Inhibitors from the Marine Sponge Penares schulzei.

    Takada K, Uehara T, Nakao Y, Matsunaga S, van Soest RWM, Fusetani N

    J. Am. Chem. Soc.   126   187 - 193  2004.01  [Refereed]

    DOI

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    105
    Citation
    (Scopus)
  • 55(P-25) Schulzeines A-C, New α-Glucosidase Inhibitors from the Marine Sponge Penares schulzei

    Takada Kentaro, Uehara Taisuke, Nakao Yoichi, Matsunaga Shigeki, Fusetani Nobuhiro

    Symposium on the Chemistry of Natural Products, symposium papers   ( 45 ) 323 - 328  2003.09

     View Summary

    α-Glucosidases not only process protein glycosylation, but also control oligosaccharide metabolism. Thus, inhibitors of α-glucosidases are potential therapeutics for the treatment of such diseases as viral diseases, cancer, and diabetes. In the course of our continuing search for potential drug leads from Japanese marine invertebrates, we found a potent α-glucosidase-inhibitory activity in the hydrophilic extract of the marine sponge Penares schulzei, whose bioassay-guided fractionation afforded three new tetrahydroisoquinoline alkaloids named schulzeines A-C. The structure of schulzeine A was elucidated by spectral analysis and chemical degradations to be the isoquinoline alkaloids, encompassing two amino acids and a C_<28> trisulfated fatty acid. The absolute stereochemistry of schulzeine A was determined by application of the advanced Mosher analysis to fragments obtained by chemical degradation. Schulzeine B was a desmethyl derivative of schulzeine A, with a stereogenic center in the tricyclic portion being epimerised. Schulzeine C was an epimer of schulzeine B. NMR data of the tricyclic part in schulzeine C was superimposable on that of schulzeine A, while the fatty acid portion was identical with that of schulzeine B. Schulzeines A-C inhibit α-glucosidase with IC_<50> values of 48-170nM. It should be noted that desulfated schulzeines also inhibited the enzyme albeit with diminished activity, thereby indicating that the activity of schulzeines was not thoroughly due to the presence of the three sulfate groups.

    DOI CiNii

  • (Z)-Sarcodictyin A, a new highly cytotoxic diterpenoid from the soft coral Bellonella albiflora

    Nakao, Y, Yoshida, S, Matsunaga, S, Fusetani, N

    J. Nat. Prod.   66   524 - 527  2003  [Refereed]

    Authorship:Lead author

  • Callysponginol sulfate A; an MT1-MMP inhibitor isolated from the marine sponge Callyspongia truncata

    Fujita, M, Nakao, Y, Matsunaga, S, van Soest, R. W. M, Itoh, Y, Seiki, M, Fusetani, N

    J. Nat. Prod.   66   569 - 571  2003  [Refereed]

  • More Kapakahines from the Marine Sponge Cribrochalina olemda

    Nakao, Y, Kuo, J, Yoshida, W. Y, Kelly, M, Scheuer, P. J

    Org. Lett.   5   1387 - 1390  2003  [Refereed]

    Authorship:Lead author

  • Ageladine A : an anti-angiogenic matrixmetalloproteinase inhibitor from the marine sponge Agelas nakamurai

    Fujita, M, Nakao, Y, Matsunaga, S, Seiki, M, Itoh, Y, Yamashita, J. van Soest, R. W. M, Fusetani, N

    J. Am. Chem. Soc.   125   15700 - 15701  2003  [Refereed]

  • Kulokekahilide-1, a cytotoxic depsipeptide from the cephalaspidean mollusk Philinopsis speciosa

    Kimura, J, Takada, Y, Inayoshi, T, Nakao, Y, Goetz, G, Yoshida, W. Y, Scheuer, P. J

    J. Org. Chem.   67   1760 - 1767  2002  [Refereed]

  • Callysponginic acid, a polyacetylenic acid which inhibits -glucosidase, from the marine sponge Callyspongia truncata

    Nakao, Y, Uehara, T, Matsunaga, S, Fusetani, N, van Soest, R. W. M

    J. Nat. Prod.   65   922 - 924  2002  [Refereed]

    Authorship:Lead author

    CiNii

  • Sodium 1-(12-hydroxy)octadecanyl sulfate, an MMP2 inhibitor, isolated from a tunicate of the family Polyclinidae

    Fujita, M, Nakao, Y, Matsunaga, S, Nishikawa, T, Fusetani, N

    J. Nat. Prod.   65   922 - 924  2002  [Refereed]

  • Nobiloside, a new neuraminidase inhibitory triterpenoidal saponin from the marine sponge Erylus nobilis

    Takada, K, Nakao, Y, Matsunaga, S, van Soest, R. W. M, Fusetani, N

    J. Nat. Prod.   65   411 - 413  2002  [Refereed]

    DOI

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    33
    Citation
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  • Dogger Bank Itch revisited: isolation of (2-hydroxyethyl) dimethylsulfoxonium chloride from the marine sponge Theonella aff. mirabilis as a cytotoxic constituent

    Warabi, K, Nakao, Y, Matsunaga, S, Fukuyama, T, Kan, T, Yokoshima, S, Fusetani, N

    Comp. Biochem. Physiol. Part B   128   27 - 30  2001  [Refereed]

  • Ancorinosides B-D, inhibitors of membrane type 1 matrix metalloproteinase (MT1-MMP), from the marine sponge Penares sollasi

    Fujita, M, Nakao, Y, Matsunaga, S, Seiki, M, Itoh, Y, van Soest, R. W. M, Fusetani, N

    Tetrahedron   57   1229 - 1234  2001  [Refereed]

    CiNii

  • Isolation and structure elucidation of two phosphorylated sterol sulfates, MT1-MMP inhibitors from a marine sponge Cribrochalina sp.: revision of the structures of haplosamates A and B

    Fujita, M, Nakao, Y, Matsunaga, S, Seiki, M, Itoh, Y, van Soest, R. W. M, Heubes, M, Faulkner, D. J, Fusetani, N

    Tetrahedron   57   3885 - 3890  2001  [Refereed]

    CiNii

  • Three new bromotyrosine derivatives lethal to crab from the marine sponge, Pseudoceratina purpurea

    Fusetani, N, Masuda, Y, Nakao, Y, Matsunaga, S, van Soest, R. W. M

    Tetrahedron   57   7507 - 7511  2001  [Refereed]

  • Calyceramides A-C, neuraminidase inhibitory sulfated ceramides from the marine sponge Discodermia calyx

    Nakao, Y, Takada, K, Matsunaga, S, Fusetani, N

    Tetrahedron   57   3013 - 3017  2001  [Refereed]

    Authorship:Lead author

    DOI

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    37
    Citation
    (Scopus)
  • Penarolide sulfates A1 and A2, new -glucosidase inhibitors from a marine sponge Penares sp.

    Nakao, Y, Maki, T, Matsunaga, S, van Soest, R. W. M, Fusetani, N

    Tetrahedron   56   8977 - 8987  2000  [Refereed]

    Authorship:Lead author

    DOI CiNii

    Scopus

    36
    Citation
    (Scopus)
  • Miraziridine A, a novel cysteine protease inhibitor from the marine sponge Theonella aff. mirabilis

    Nakao, Y, Fujita, M, Warabi, K, Matsunaga, S, Fusetani, N

    J. Am. Chem. Soc.   122   10462 - 10463  2000  [Refereed]

    Authorship:Lead author

  • Pseudotheonamides, serine protease inhibitors from the marine sponge Theonella swinhoei

    Nakao, Y, Masuda, A, Matsunaga, S, Fusetani, N

    J. Am. Chem. Soc.   121   2425 - 2431  1999  [Refereed]

    Authorship:Lead author

  • Koshikamide A1, a new cytotoxic linear peptide isolated from the marine sponge, Theonella sp.

    Fusetani, N, Warabi, K, Nogata, Y, Nakao, Y, Matsunaga, S

    Tetrahedron Lett.   40   4687 - 4690  1999  [Refereed]

    CiNii

  • Tokaramide A, a new cathepsin B inhibitor from the marine sponge Theonella aff. mirabilis

    Fusetani, N, Fujita, M, Nakao, Y, Matsunaga, S

    Bioorg. Med. Chem. Lett.   9 ( 24 ) 3397 - 3402  1999  [Refereed]

    DOI

  • More peptides and other constituents of the marine mollusk Philinopsis speciosa

    Nakao, Y, Yoshida, W. Y, Szabo, C. M, Baker, B. J, Scheuer, P. J

    J. Org. Chem.   63   3272 - 3280  1998  [Refereed]

    Authorship:Lead author

    CiNii

  • Isolation of 1-methylherbipoline salts of halisulfate-1 and suvanine as serine protease inhibitors from a marine sponge, Coscinoderma mathewsi

    Kimura, J, Ishitsuka, E, Nakao, Y, Yoshida, W. Y, Scheuer, P. J, Kelly-Borges, M

    J. Nat. Prod.   61   248 - 250  1998  [Refereed]

  • Cyclotheonamides E2 and E3, new potent serine protease inhibitors from the marine sponge of the genus Theonella

    Nakao, Y, Oku, N, Matsunaga, S, Fusetani, N

    J. Nat. Prod.   61   667 - 670  1998  [Refereed]

    Authorship:Lead author

  • Two acyclic kahalalides from the sacoglossan mollusk Elysia rufescens

    Goetz, G, Nakao, Y, Scheuer, P. J

    J. Nat. Prod.   60   562 - 567  1997  [Refereed]

  • Cupolamide A: a cytotoxic cyclic heptapeptide from two samples of the sponge Theonella cupola

    Bonnington, L. S, Tanaka, J, Higa, T, Kimura, J, Yoshimura, Y, Nakao, Y, Yoshida, W. Y, Scheuer, P. J

    J. Org. Chem.   62 ( 22 ) 7765 - 7767  1997  [Refereed]

     View Summary

    A new cyclic heptapeptide cupolamide A (1) was isolated from two samples of the sponge Theonella cupola collected in Indonesia and Okinawa. It contains two L-Val, one D-Leu, and one D-Ser, and three uncommon amino acid residues, D-homoarginine (Har), trans-4-hydroxy-L-proline (Hyp), and L-2,4-diaminobutanoic acid (Dba). The structure elucidation of 1 was performed by NMR spectroscopic techniques and the stereochemistry was determined by Marfey's method. Cupolamide A (1) was active against P388 murine leukemia cells with an IC50 = 7.5 μg/mL.

    DOI

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    35
    Citation
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  • Two diverse constituents of the cephalaspidean mollusk Smaragdinella calyculata

    Szabo, C. M, Nakao, Y, Yoshida, W. Y, Scheuer, P. J

    Tetrahedron   52   9681 - 9686  1996  [Refereed]

  • Waiakeamide, a cyclic hexapeptide from the sponge Ircinia dendroides

    Mau, C. M. S, Nakao, Y, Yoshida, W. Y, Scheuer, P. J

    J. Org. Chem.   61   6302 - 6304  1996  [Refereed]

  • The kapakahines, cyclic peptides from the marine sponge Cribrochalina olemda

    Yeung, B. K. S, Nakao, Y, Kinnel, R. B, Carney, J. R, Yoshida, W. Y, Scheuer, P. J

    J. Org. Chem.   61   7168 - 7173  1996  [Refereed]

  • Kulolide, a cytotoxic depsipeptide from a cephalaspidean mollusk, Philinopsis speciosa

    Reese, M. T, Gulavita, N. K, Nakao, Y, Hamann, M. T, Yoshida, W. Y, Coval, S. J, Scheuer, P. J

    J. Am. Chem. Soc.   118   11081 - 11084  1996  [Refereed]

  • Pupukeamide, a linear tetrapeptide from a cephalaspidean mollusk Philinopsis speciosa

    Nakao, Y, Yoshida, W. Y, Scheuer, P. J

    Tetrahedron Lett.   37   8993 - 8996  1996  [Refereed]

    Authorship:Lead author

    CiNii

  • Three more cyclotheonamides, C, D, and E, potent thrombin inhibitors from the marine sponge Theonella swinhoei

    Nakao, Y, Matsunaga, S, Fusetani, N

    Bioorg. Med. Chem.   3   1115 - 1122  1995  [Refereed]

    Authorship:Lead author

  • Kumusine, a chloroadenine ribosidefrom a sponge, Theonella sp.

    Ichiba, T, Nakao, Y, Scheuer, P. J, Sata, U. S, Kelly-Borges, M

    Tetrahedron Lett.   36   3977 - 3980  1995  [Refereed]

  • Kapakahine B: a cyclic hexapeptide with an alpha-carboline ring system from the marine sponge Cribrochalina olemda

    Nakao, Y, Yeung, B. K. S, Yoshida, W. Y, Scheuer, P. J, Kelly-Borges, M

    J. Am. Chem. Soc.   117   8271 - 8272  1995  [Refereed]

    Authorship:Lead author

  • Axinelloside A, an unprecedented highly sulfated lipopolysaccharide inhibiting telomerase, from the marine sponge, Axinella infundibula

    Nakao, Y, Matsunaga, S, Fusetani, N

    Tetrahedron Lett.   34   1511 - 1514  1993  [Refereed]

    Authorship:Lead author

  • Toxadocials B, C and Toxadocic acid A: thrombin-inhibitory aliphatic tetrasulfates from the marine sponge, Toxadocia cylindrica

    Nakao, Y, Matsunaga, S, Fusetani, N

    Tetrahedron   49   11183 - 11188  1993  [Refereed]

    Authorship:Lead author

  • Eight new cytotoxic metabolites closely related to onnamide A from two marine sponges of the genus Theonella

    Matsunaga, S, Fusetani, N, Nakao, Y

    Tetrahedron   39   8369 - 8376  1992  [Refereed]

  • Nazumamide A, a thrombin-inhibitory tetrapeptide, from a marine sponge, Theonella sp.

    Fusetani, N, Nakao, Y, Matsunaga, S

    Tetrahedron Lett.   32   7073 - 7074  1991  [Refereed]

▼display all

Books and Other Publications

  • 理工系のための一般化学

    柴田, 高範, 石原, 浩二, 井村, 考平, 鹿又, 宣弘, 寺田, 泰比古, 中井, 浩巳, 中尾, 洋一, 中田, 雅久, 古川, 行夫, 山口, 正( Part: Contributor, pp 174-184)

    東京化学同人  2021.01 ISBN: 9784807909940

  • Handbook of marine natural products

    Yoichi Nakao, Nobuhiro Fusetani( Part: Contributor)

    Springer  2012.04 ISBN: 9789048138333

  • マリンバイオテクノロジーの新潮流

    児玉公一郎, 中尾洋一( Part: Contributor, pp 74-92)

    シーエムシー出版  2011.11 ISBN: 9784781304533

  • マリンバイオテクノロジー : 海洋生物成分の有効利用

    中尾洋一( Part: Contributor, pp 82-96)

    シーエムシー出版  2010.09 ISBN: 9784781302676

  • Comprehensive natural products II : chemistry and biology

    Nakao, Yoichi, Fusetani, Nobuhiro( Part: Contributor)

    Elsevier  2010.05 ISBN: 9780080453811

  • 海洋生物成分の利用 : マリンバイオのフロンティア

    中尾洋一( Part: Contributor, pp 82-96)

    シーエムシー出版  2005.03 ISBN: 4882314940

▼display all

Presentations

  • Anti-parasitic agents from marine organisms

    Nakao, Y

    Presentation date: 2020.09

  • ココア主要成分テオブロミンの新たな生理作用

     [Invited]

    Presentation date: 2019.09

  • ケミカルエピゲノミクスによる天然化合物の探索

    中尾洋一  [Invited]

    Presentation date: 2019.03

  • 機能性味噌とエピゲノム調節

    中尾洋一  [Invited]

    Presentation date: 2018.10

  • 味噌に含有する抗ストレス物質の探索及びその成分を強化したみその製法について

    中尾洋一  [Invited]

    Presentation date: 2018.10

  • 医療のさらなる安心・安全を志向する若手研究者のイノベーティブチャレンジ 時計遺伝子Per2の発現周期やマウス行動周期に対する遠志の影響について

    原口 敦嗣, 中村 文彬, 阿部 真太郎, 中尾 洋一, 柴田 重信

    日本薬学会年会要旨集  (公社)日本薬学会

    Presentation date: 2018.03

    Event date:
    2018.03
     
     
  • 共生微生物によって生合成される海洋天然化合物

     [Invited]

    Presentation date: 2017.09

  • 味噌に含まれる抗ストレス成分の探索

     [Invited]

    Presentation date: 2017.03

  • 味噌に含まれる抗ストレス有効成分とその生成過程における発酵の意義

     [Invited]

    Presentation date: 2017.01

  • 既知天然化合物の再発見

     [Invited]

    Presentation date: 2016.11

  • 発酵食品とエピゲノム情報

     [Invited]

    Presentation date: 2016.07

  • New approach to study histone modification changes controlling biological phenomena by small molecules

    Nakao, Y, Arai, D, Kina, Y, Kataoka, R, Hayashi-Takanaka, Y, Kimura, H  [Invited]

    Presentation date: 2015.12

  • 海洋毒・海洋生理活性物質の探索研究へのMSの応用

     [Invited]

    Presentation date: 2015.06

  • Chemical biology of cell differentiation using chemical probes derived from marine natural products

    Otsuka, S, Iwata, T, Tsubokura, K, Arai, D, Fukase, K, Tanaka, K, Nakao, Y

    Presentation date: 2014.11

  • Search for natural products controlling epigenetics

    Nakao, Y  [Invited]

    Presentation date: 2014.10

  • 天然化合物によるエピゲノム制御

     [Invited]

    Presentation date: 2014.06

  • 味噌成分のクロマチン修飾におよぼす生理活性プロファイルの解析

     [Invited]

    Presentation date: 2014.06

  • 環境エピゲノム研究におけるケミカルバイオロジー

     [Invited]

    Presentation date: 2013.06

  • Shinanthrene Aの立体化学研究と生物活性

    町田光史, 岡本真由美, 清水功雄, 中尾洋一

    日本化学会講演予稿集 

    Presentation date: 2013.03

    Event date:
    2013.03
     
     
  • 新規天然化合物shinanthrene Aとそのエストロゲン活性

    町田光史, 岡本真由美, 清水功雄, 新井大祐, 伏谷伸宏, 中尾洋一

    日本分子生物学会年会プログラム・要旨集(Web) 

    Presentation date: 2013

    Event date:
    2013
     
     
  • Anti-parasitic agents from marine organisms

    Nakao, Y  [Invited]

    Presentation date: 2012.11

  • DNA損傷機構に影響する化合物の探索

     [Invited]

    Presentation date: 2011.07

  • Anti-protozoan Agents from Marine Invertebrates

    Nakao ,Y, Ishigami, S, Goto, Y, Kawazu, S, Matsumoto, Y  [Invited]

    Presentation date: 2010.12

  • 海洋天然物化学の来し方行く末

     [Invited]

    Presentation date: 2010.07

  • 医薬品素材としての海洋天然化合物

     [Invited]

    Presentation date: 2010.03

  • 天然化合物を利用したケミカルエピゲノミクス研究の可能性について

     [Invited]

    Presentation date: 2009.06

  • Marine Natural Products as Tool for Chemical Biology

    Nakao, Y  [Invited]

    Presentation date: 2009.03

  • Search for Chemical-“Epi”genetic Probes from Marine Invertebrates

    Nakao, Y  [Invited]

    Presentation date: 2009.03

  • Searching Probe molecules for Chemical-Epigenetics from Marine Invertebrates

    Nakao, Y  [Invited]

    Presentation date: 2008.12

  • 幹細胞のケミカルバイオロジー

     [Invited]

    Presentation date: 2008.04

  • Bioactive small molecules from marine organisms

    Nakao, Y  [Invited]

    Presentation date: 2007.09

  • 海洋生物からの血管新生阻害剤の探索

     [Invited]

    Presentation date: 2007.05

  • 海洋天然物化学とケミカルバイオロジー

     [Invited]

    Presentation date: 2006.05

  • Anti-angiogenic agents from marine invertebrates

    Nakao, Y  [Invited]

    Presentation date: 2005.12

  • Application of target- and phenotype-oriented bioassays to the search program for anti-angiogenic agents from marine organisms

    Nakao, Y  [Invited]

    Presentation date: 2005.09

  • 水産生物の保健機能素材としての可能性

     [Invited]

    Presentation date: 2005.06

  • 海洋生物由来血管新生阻害剤の探索

     [Invited]

    Presentation date: 2005.03

  • 海洋生物からの血管新生阻害剤の探索

     [Invited]

    Presentation date: 2005.02

  • 生命現象を探るツールを海洋生物に求める

     [Invited]

    Presentation date: 2004.11

▼display all

Research Projects

  • Elucidation of mechanisms for macrophage manipulation by Leishmania

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2022.05
    -
    2025.03
     

  • Elucidation of the regulatory mechanism of feline mammary carcinoma by integrating three-dimensional culture method and pharmacokinetic analysis

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2021.04
    -
    2025.03
     

  • エピゲノム修飾をマーカーとした腸内フローラにおける糖尿病合併症制御因子の探索

    日本学術振興会  科学研究費助成事業

    Project Year :

    2022.06
    -
    2024.03
     

    中山 二郎, 井上 貴子, 中尾 洋一

  • Integrated platform for analyzing the modes of actions of natural products

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2021.04
    -
    2024.03
     

  • International collaborative study on foods, microbiota, and life style diseases of Asians

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2020.10
    -
    2024.03
     

  • Understanding microbial uncultivability: cue for the growth and incapability of colony formation

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)

    Project Year :

    2020.07
    -
    2023.03
     

  • Studies of feline morbillivirus pathogenesis

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

    Project Year :

    2020.04
    -
    2023.03
     

  • Leishmania parasites as biological resources for immunomodulatory molecules

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)

    Project Year :

    2020.07
    -
    2022.03
     

    Goto Yasuyuki

     View Summary

    Leishmania parasites proliferate within macrophages (Mφ) in the mammalian host, and they need to manipulate Mφ to ensure its efficient survival. In this study, we focused on the ability of Leishmania to regulate Mφ and aimed to identify the protozoan factors responsible for this regulation. We conducted our research by establishing screening methods, fractionating protozoan extracts, and identifying active substances. In FY2020 we succeeded to establish various screening assays, and in FY2021 fractionation-based search of active compounds were performed using the mentioned assay systems. As a result of screening, several candidate molecules were identified, but when recombinant forms of them were produced, they did not show desired activities. Therefore, we decided to improve the assays for further screening. Some active fractions were determined by the new assay, whereas we could not reach to identification of active molecules.

  • Cultivation of unculitivated microorgasnisms and clarification of growth controling mechanisms by focusing on microbial interactions

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2019.04
    -
    2022.03
     

    Aoi Yoshiteru

     View Summary

    Over 99% of microorganisms in environment remain uncultured. However, very few attempts have been carried out for development of new microbial cultivation technology, and reason why most microorganisms resist cultivation has not been clarified so far. The objective of the study is to establish and apply new cultivation methods based on a new concept, and to discover previously unknown mechanism controlling growth of uncultivated microorganisms. In this study, first, we successfully applied several new cultivation methods to environmental sample for isolating bacteria. Methods are in situ cultivation, a fully automated microbial cultivation method, and Gel-Micro-Droplet (GMD)-aggregate cultivation (a new microbial cultivation platform providing extremely high inoculum cell density). Second, through testing isolates from above methods, it was shown that the microbial interactions clearly affect their growth.

  • Search for small molecules controlling cell differentiation

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2018.04
    -
    2021.03
     

    NAKAO YOICHI

     View Summary

    In this research project, 3D culture system monitoring multiple cell differentiation pathways has been established. Screening for bioactive food ingredients using this system has been conducted. As the result, 48 out of 96 food samples showed some kinds of activities. The most active food sample was selected for further study to isolate active substances. The isolation is at the final step currently. Structure elucidation of the bioactive subtances will be conducted in the nearest future.

  • なぜ多くの微生物は培養困難なのか?未知増殖制御メカニズムの発見と解明

    日本学術振興会  科学研究費助成事業 挑戦的研究(萌芽)

    Project Year :

    2018.06
    -
    2020.03
     

    青井 議輝, 中尾 洋一

  • Establishment and re-establishment of uterine environment for pregnancy success

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

    Project Year :

    2016.04
    -
    2020.03
     

    Imakawa Kazuhiko

     View Summary

    In ruminants, trophoblast cells go through epithelial-mesenchymal transition (EMT) when implanting to the maternal endometrium. As EMT begins, trophoblast cells fuse each other to generate bi-nucleate or multi-nucleated cells, from which pregnancy associated glycoprotein is released. In addition, exosomes, miRNA and lncRNA were found from uterine flushing media and their functions were examined using in vitro uterine epithelial mono-culture or uterine epithelial-trophoblast co-culture systems. miRNA and lncRNA are effective in controling maternal immune system for semi-allogenic trophoblast cells.

  • Detection of chemical substances with epigenetic activity to protect environmental risk by the adverse outcome pathway approach

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

    Project Year :

    2015.04
    -
    2019.03
     

    Sone Hideko, Nansai Hiroko, Kondo Masayo, Suzuki Shinnosuke, Otsuka Satoshi

     View Summary

    We developed a method to visually detect DNA methylation and histone modification as an index of epigenetic status using pluripotent stem cells like mouse ES and human iPS cells. Using this engineered cell lines, one hundred and thirty-five chemical substances were analyzed. Further epigenetic toxicity was detected by examining DNA methylation variation in the promoter region of the disease causing gene in detail by analysis of specific DNA sequences. The effect of irradiation on the differentiation of human iPS cells into retinal ganglion cells was also investigated. Further we have developed a method that can detect differences in epigenetic status during differentiation derived from undifferentiated stem cells. These results suggest that the rapid detection of epigenetic activity using pluripotent stem cells can detect initial events of exposures to chemicals and environmental factors and then predict the later effects.

  • Development of novel methods for target identification of natural products and chemical epigenetics

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (S)

    Project Year :

    2014.05
    -
    2019.03
     

    Yoshida Minoru

     View Summary

    We have developed a unique platform to systematically identify target molecules of bioactive compounds by utilizing novel technologies such as molecular barcode sequencing and bimolecular luminescence complementation. Specifically, an integrated method to detect chemical genomic interaction using barcoded gene disruptant libraries of both budding and fission yeasts, a barcoded, pooled shRNA viral library, and a split luciferase-based mammalian three-hybrid system was established. By using this method as well as a conventional physical interaction screening using compound-conjugated beads, we could identify target molecules and the mechanisms of action of several target-unknown metabolites from microbes and marine organisms.

  • Activation of protoplast function by nanobubbles and development of high-speed production system for useful metabolites

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2015.04
    -
    2018.03
     

    AOYAGI Hideki, BAMBA Takeshi, NAKAO Yoichi, OU Hekisyou

     View Summary

    The nanobubble generation system was originally developed. This system can generate nanobubbles of various gases with high efficiency. The generated nanobubbles could stably exist in different solutions and media. Various investigations using yeast, plants cells and protoplasts suggested that the metabolites biosynthesized by protoplasts were released into the culture broth, with the double benefits of increasing the overall productivity and facilitating downstream processing. Various nanobubbles enhance those phenomena. Application of nanobubbles to yeast, various plants cells and protoplasts resulted in efficient production of useful metabolites. The bioreactor capable of supplying nanobubbles was also developed. The nanobubbles can be also used for ripening of fruit, inactivation of microbes, removal of chemicals on fruit surface, etc

  • Euro-Japan collaborative field work for the understanding of symbiotic evolution of marine microbes

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2013.04
    -
    2016.03
     

    Nakao Yoichi, YAMASHITA Jun, KIMURA Hiroshi, IKURA Tsuyoshi

     View Summary

    We have compared the metabolome analysis of marine sponge by LC-MS/MS. As the results, we have found the difference between the matabolite profiles of two closely related sponge samples collected in Italy and Japan. Also, we could isolate highly novel microorganisms from the sponge of genus Theonella by means of newly developed Diffusion Chamber method. Through this project, a sound base for the collaborative activity between Italy-Japan marine natural products chemistry has been established.

  • エピゲノム情報に基づく日本食がストレスに与える影響の評価

    農林水産省  核心的技術創造促進事業 異分野融合共同研究 医学・栄養学との連携による日本食の評価 世界の健康に貢献する日本食の科学的・多面的検証

    Project Year :

    2014
    -
    2016
     

    中尾洋一

  • Analysis of stem cell differentiation process on the basis of epigenetic change

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

    Project Year :

    2013.04
    -
    2014.03
     

    NAKAO Yoichi, KIMURA Hiroshi, YAMASHITA Jun

     View Summary

    In this study, we have screened activities causing selective histone modification using the crude extracts library prepared from marine organisms. We have searched for active substances from the hit samples and evaluated their effects on the stem cell differentiation.
    As the result of the screening, many hit samples showed up. We have isolated psammaplin A, known as an inhibitor against HDAC, as the active substance causing H3K9 acetylation. Psammaplin A induced differentiation to mesoderm and endoderm in the process of the formation of embryoid body. On the other side, an active fraction, showing strong inhibition against HDAC1 and differentiation to neural cells, was obtained from the marine sponge Mycale sp.

  • Chemical Biology on the Symbiotic System in Marine Invertebrates

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2012.04
    -
    2013.03
     

    NAKAO Yoichi, KIMURA Nobutada

     View Summary

    It has been disclosed that most of the marine secondary metabolites originate from the symbiotic microbes. In this research project, we focused on an unique marine metabolite, calyculin A from a marine sponge Discodermia calyx, and tried to separate and identify the calyculin A producing microbe. We have succeeded to obtain the fraction with concentrated microbes thought to produce calyculin A and extraction of genomic DNA from this assembly of the concentrated microbes. The metagenomic DNA sequencing analysis of the extracted genomic DNA and the following DNA mapping allowed us to identify the major microbe constituents and 13 contigs of PKS gene clusters

  • Chemical Biology on the Symbiotic System in Marine Invertebrates

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

    Project Year :

    2012
     
     
     

    NAKAO Yoichi, KIMURA Nobutada

     View Summary

    It has been disclosed that most of the marine secondary metabolites originate from the symbiotic microbes. In this research project, we focused on an unique marine metabolite, calyculin A from a marine sponge Discodermia calyx, and tried to separate and identify the calyculin A producing microbe. We have succeeded to obtain the fraction with concentrated microbes thought to produce calyculin A and extraction of genomic DNA from this assembly of the concentrated microbes. The metagenomic DNA sequencing analysis of the extracted genomic DNA and the following DNA mapping allowed us to identify the major microbe constituents and 13 contigs of PKS gene clusters.

  • Research for useful marineorganisms in Asia and Pacific regions

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2009
    -
    2011
     

    NAKAO Yoichi, HORI Kanji, SAKAI Ryuichi, FUSETANI Nobuhiro, MATSUOKA Shunji, FUKUZAWA Seketsu, WAKIMOTO Toshiyuki, FUJITA Masaki, TAKADA Kentarou

     View Summary

    In this project, a total 515 marine organisms were collected from 3 countries and preserved as the extract-, microbe-, and genome-libraries. Currently, evaluation for bioactivities of these extracts are being carried out. At the same time, thight collaborative relationship among the researchers of these countries and our group was founded, through this project.

  • Chemical Biology of Stem cell differentiation

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2007
    -
    2009
     

    NAKAO Yoichi

     View Summary

    We were planning to find small molecules useful for the study of the controlling mechanism of stem cell differentiation to vascular systems. We have studied application of marine natural anti-angiogenic compounds, azumamides, to the in vitro vascular re-organizing model using mouse iPS cells, in which we confirmed their potent effects. Therefore, this model, as well as that using mouse ES cells, was shown to be useful for the study of stem cell differentiation systems. Next, we have evaluated anti-angiogenic effects of analogues of geladine, in which we could identified unknown targets for ageladines besides the known MMP.

  • 海洋無脊椎動物からのガン関連酵素を標的とする機能分子の探索

    日本学術振興会  科学研究費助成事業 特定領域研究

    Project Year :

    2004
    -
    2007
     

    松永 茂樹, 中尾 洋一

     View Summary

    1.大島新曽根産海綿Pseudoceratina purpurea由来の変異株酵母に対する生育阻害物質同海綿の抽出物を、液液分配、逆相カラムクロマトグラフィーおよび逆相液体クロマトグラフィーに付して、pseudoceratin AおよびBと命名した活性物質を単離した。これらの化合物は互いにジアステレオ異性体の関係にある、臭素を4つ含む化合物であった。NMRデータの解析から5つの部分構造を明らかにし、さらにHMBCデータからそれらの配列順を決定した。この段階で3つのエーテル結合の配置が不明であったため、NMRデータの詳細な解析ならびに化学シフトおよび結合定数の文献値との比較などにより、平面構造の結論を得た。6員環の周りの相対配置はNOEデータから決定し、鎖状部に存在する3連続した2級アルコールの相対配置もNMRデータから導いた。2,3,4-トリヒドロキシ-1,5-ペンタンジアミン部の絶対配置は、加水分解によって取得した当該フラグメントのビスMTPAアミドのNMRデータを合成標品の物と比べることにより、D-アラビトールと同じ配置であることを明らかにした。
    2.大島新曽根産海綿Jaspis serpentina由来の細胞毒性物質同海綿の抽出物から、液液分配および逆相液体クロマトグラフィーなどにより、2つの活性化合物を得た。ひとつ目の化合物は既知化合物のpoecillastrin Cであったが、二つ目の化合物は類縁の新しい化合物であったため、poecillastrin Dと命名した。この化合物の平面構造を、機器分析データの解析により明らかにした。

  • ALPHASCREENによるソフトな膜タンパク質間相互作用の解析法開発

    日本学術振興会  科学研究費助成事業 特定領域研究

    Project Year :

    2004
     
     
     

    中尾 洋一

     View Summary

    平成16年度の研究では、がん細胞の浸潤や血管新生における血管内皮細胞の遊走に必須であるMMP2/TIMP2/MT1-MMPの安定複合体と接着分子CD44との間のソフトな相互作用を検出するために、ALPHASCREENを用いたソフトな相互作用検出法を開発し、この相互作用を阻害する低分子性化合物を探索することを目的として研究を行った。今年度の研究成果の概要を以下に述べる。
    MT1-MMPおよびCD44に対して、当初はアクセプターおよびドナービーズに固定したそれぞれの抗体を介してタンパク質間の相互作用を検出する予定であったが、3カ所での相互作用の形成(抗体1/MT1-MMP/CD44/抗体2)が必要となるため、検出感度が悪く相互作用の検出ができなかった。また、MT1-MMPは複合体を形成させている間に自己消化を起こしてしまい、バラバラのフラグメントになったものでは相互作用が起きないことも明らかになった。そこで、自己消化を防ぐためにMMP阻害剤をバッファー中に添加し、さらにCD44をドナービーズに直接固定化することで相互作用を2カ所に減らしたところ、384穴マイクロプレートを用いた小スケールアッセイにおいて、タンパク質の検出感度1pmol(100-200ng/サンプル)程度で相互作用を検出することに成功した。
    また、CD44のステム領域とMT1-MMPの結合部位(ヘモペキシン様ドメイン:PEX)を、それぞれ直接固定したビーズを用いて、CD44とMT1-MMPの相互作用をダイレクトに検出する高感度アッセイの検討を行っている。アッセイに用いるPEXを供給するため、大腸菌を用いて組み換えPEXの発現系を導入した。なお、組み換えPEXは分子量が2万程度でNMR測定が可能となるため、今年度中に^<15>Nでラベルしたタンパク質を用いてNMRによる相互作用の検証を行う予定である。
    一方、本アッセイで用いるALPHAビーズは蛍光顕微鏡下で観察可能なことから、抗MMP抗体コートしたALPHAビーズをヒト子宮がん由来HeLa細胞培養液中に添加したところ、MMPが局在している細胞の浸潤突起に結合している様子が観察された。そこで、ALPHAビーズと蛍光タイムラプスイメージングを組み合わせて、MMP2/TIMP2/MT1-MMP複合体の細胞表面上での挙動を一分子単位で観察できるシステムの構築を検討中である。さらに、共焦点レーザー顕微鏡を用いて、680nmのレーザー光をあてると相互作用を起こしているALPHAビーズだけが蛍光を発することを確認できたため、生細胞表面で起きている膜タンパク質間相互作用の検出法についても検討中である。

  • 化学遺伝学的手法による血管新生阻害剤の探索

    日本学術振興会  科学研究費助成事業 特定領域研究

    Project Year :

    2004
     
     
     

    中尾 洋一

     View Summary

    平成16年度は血管新生関連酵素に対する阻害活性試験と、マウスES細胞由来Flk1^+細胞からのin vitro血管再構築系による化学遺伝学的手法により、海洋無脊椎動物を対象とした血管新生阻害剤探索とその作用機序解明を行った。
    この結果、3種の化合物に非常に有望な血管新生阻害活性を認めた。これら3種はそれぞれ、MMP、HDAC、VEGFRのチロシンキナーゼと標的分子が異なることから、本研究によって幅広い作用機構の血管新生阻害剤を見出すことが出来た。
    このうち、MMP阻害剤として得られた既知化合物avaroneは、その後血管内皮増殖因子受容体(VEGFR)Flt1のチロシンキナーゼ選択的阻害、HUVECに対する遊走阻害、in vivoでの血管新生阻害、および抗腫瘍活性等の有望な活性が認められたため、血管新生阻害剤として、スクリーニング委員会と共同で出願特許申請を行った。
    なお、血管新生阻害活性を示す新規MMP阻害剤ageladine Aについてはin vivo抗腫瘍活性をスクリーニング委員会にて検定中である。
    遺伝子発現に関わる酵素であるヒストンデアセチラーゼ(HDAC)の阻害剤は血管新生を阻害することが知られている。申請者により単離された新規HDAC阻害剤azumamide Aは、ES細胞を用いた血管再構築系において調べたところ、血管新生を阻害することから、in vivoの血管新生阻害試験を検討中である。

  • 部位特異的MT1-MMP阻害剤の探索

    日本学術振興会  科学研究費助成事業 若手研究(A)

    Project Year :

    2002
    -
    2004
     

    中尾 洋一

     View Summary

    平成16年度の本研究では、1種の海洋無脊椎動物サンプルから、3種の新規MMP2およびMT1-MMP阻害剤を得た(投稿準備中)。また、昨年度の研究で見出した新規MMP2阻害剤ageladine Aは、既存のMMP阻害剤とは全く異なる新しいタイプの阻害剤であることが期待されているため、X線結晶解析により阻害機構を明らかにするべく、結晶化に最適の組み換えタンパク質の作成を試みている。
    また、Alphascreen(TM)を用いた、タンパク質-タンパク質相互作用阻害アッセイ法の構築を引き続き行い、組み換えCD44分子とMT1-MMP分子を用いて1pmol程度で相互作用を検出することに成功した。現在、MT1-MMPの相互作用部位であるPEXドメインのみを用いて、さらに高感度検出が可能なアッセイ系の立ち上げを行っている。
    さらに、細胞表面に発現しているCD44およびMT1-MMPを標的にした抗体コートしたAlphaビーズを培養細胞に投与し準ところ、細胞の浸潤突起上に結合したビーズが認められたため、生細胞表面上で実際に形成されている複合体の形成および一分子観察を標的にしたアッセイ系の開発も試みている。

  • Efficient Utilization of Fisheries Biproducts. Exploration of miomedial Resoures from Deep-Sea Spong

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2002
    -
    2003
     

    FUSETANI Nobuhiro, NAKAO Yoichi, MATSUNAGA Shigeki

     View Summary

    In order to explore the possibility of discovering new compounds with biomedical importance from deep-sea sponges, we collected 78 samples at Shinsone, near Oshima Island and examined the biological activities of their extracts. More than 70% of the extracts exhibited either cytotoxic, antimicrobial, or enzyme inhibitory activity, demonstrating the importance of deep-sea sponges in natural products drug discovery. We have isolated the following compounds from the collected specimens.
    1.Theopederin K from Discodermia japonica
    2.Shinsonefuran from Stoeba extensa
    3.Novel macrolides from Poecillastra sp.
    Isolation of cytotoxic constituents from another deep-sea sponge collected off Heda, Izu, is in progress.

  • 海洋無脊椎動物からの血管新生阻害剤の探索

    日本学術振興会  科学研究費助成事業 特定領域研究

    Project Year :

    2002
     
     
     

    中尾 洋一

     View Summary

    新たな抗がん剤探索を目的としたこれまでの研究で、血管内皮細胞の管腔形成に対する阻害活性およびマトリックスメタロプロテアーゼ(MMP)に対する阻害活性を指標として、海洋無脊椎動物を対象に血管新生阻害物質の探索を行い、6種の海綿と1種の腔腸動物から6つの新規化合物を含む11個の活性化合物を得ている。今年度の研究では新たに1種のホヤと4種の海綿から2種の既知化合物と6種の新規化合物をMMP阻害剤として得た。
    また、今年度から新たにマウスES細胞in vitro血管再構築系を用いた血管新生阻害活性試験を導入してスクリーニングを行うとともに、これまでに得られた有望活性化合物について血管新生阻害活性を調べ2つの化合物に活性を認めた。このうち1つの化合物については、スクリーニング委員会が行うヒトがん細胞パネル試験においてCOMPARE marginalと判定されユニークな作用機構が示唆されたため、Nude mouse/Xenograftによる抗腫瘍活性検定試験を準備中である。
    さらに、Flt-1のチロシンキナーゼ阻害活性およびHUVECの系における細胞遊走阻害活性が明らかとなった化合物avaroneについて、スクリーニング委員会においてNude mouse/Xenograftによるin vivo抗腫瘍活性試験を行い、腫瘍の増殖抑制が認められた。

  • 海洋無脊椎動物からの血管新生阻害剤の探索

    日本学術振興会  科学研究費助成事業 特定領域研究(C)

    Project Year :

    2001
     
     
     

    中尾 洋一

     View Summary

    海洋無脊椎動物の抽出物、について管腔形成阻害および酵素阻害アッセイを行った。酵素は血管新生に深く関与するとされている膜型マトリックスメタロプロテアーゼ1(MT1-MMP)およびマトリックスメタロプロテアーゼ2(MMP2)を用い、阻害物質の選択性もあわせて検討した。その結果、三重県五ヶ所湾産の末同定ホヤおよび鹿児島県口之永良部島産の末同定海綿に強力かつ選択的なMMP2阻害活性が認められた。
    冷凍保存されていたホヤ検体800gを抽出後、MM2阻害活性を指標に溶媒分画、各種クロマトグラフィーを用いて阻害物質を単離した。阻害物質の構造は各種スペクトルデータからその平面構造を新規化合物である12-hydroxystearyl-1-sulfateと決定した。絶対立体化学は改良Mosher法および合成標品との比較より、鏡像異性体の混合物であることが明らかになった。新規MMP2阻害物質12-hydroxystearyl-1-sulfateはMMP2をIC_<50>値9.0μg/mLで阻害した。
    鹿児島産海綿を有機溶媒で抽出後、生物活性を指標に分画をすすめ、鮮やかな青色蛍光を示す物質を活性本体として5.0mg単離した。阻害物質の分子量は356と決定されたが、プロトンNMRにおいて低磁場領域に3つのシグナルを与えるのみで、スペクトル解析による構造決定は困難であった。現在化学構造を検討中である。

  • 海洋無脊椎動物からの血管新生阻害剤の探索

    日本学術振興会  科学研究費助成事業 特定領域研究(C)

    Project Year :

    2000
     
     
     

    中尾 洋一, 松永 茂樹

     View Summary

    新しいがん治療薬の開発を目指して、海洋無脊椎動物を対象に血管新生阻害物質の探索を行った。
    本年度の研究では、血管内皮細胞の管腔形成に対する阻害活性、および内皮細胞の遊走に関与する膜型マトリックスメタロプロテアーゼ(MT1-MMP)に対する阻害活性を指標としてスクリーニングを行い、浮かび上がったサンプルから活性本体の単離と構造決定を行った。
    1.管腔形成阻害活性化合物
    2種の鹿児島県産海綿から既知の4化合物hexa-、hepta-、octaprenyl hydroquinone、およびjaspamide Aを得た。これらの化合物はそれぞれ、1.0-4.0および0.1ug/mLの濃度で管腔形成阻害活性を示した。
    2.MT1-MMP阻害剤
    鹿児島県産海綿から活性物質として既知化合物ancorinoside Aとともに3種の新規誘導体ancorinosides B-Dを単離構造決定した。これらはMT1-MMPに対してIC_<50>180-500ug/mLの阻害活性を示した。さらに、ancorinosides Bは20ug/mLの濃度で管腔形成阻害活性を示した。
    高知県産海綿から活性物質として2種の化合物、haplosamate Aとその新規誘導体を単離構造決定した。これらはMT1-MMPに対してそれぞれIC_<50>150および160ug/mLの阻害活性を示した。
    既知化合物としては、静岡県産海綿からhalistanol sulfate(IC_<50>1.0ug/mL)を、また鹿児島県産海綿からはavaroneおよびavarol(IC_<50>0.5および10ug/mL)をMT1-MMP阻害剤として単離した。
    本年度の研究を通して得られた有望な化合物については、文部省がん特定総合がん・制がん剤のスクリーニング委員会にin vivoでの抗血管新生阻害活性試験を依頼中である。

  • Development of Biochemical Reagents to Study Signal Transduction from Awuatic Organisms

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B).

    Project Year :

    1999
    -
    2000
     

    FUSETANI Nobuhiro, OZAKI Hiroshi, MATSUNAGA Shigeki, WATABE Shugo, KAMIMORI Hitoshi, NAKAO Yoichi

     View Summary

    We have collected marine invertebrates along the coast of Japan and carried out screening for biological activities. Based on the results of the screening we have isolated and characterized the active principles and studied their modes of action, which may lead to the development of biochemical reagents.
    1. Isolation and structure elucidation of novel bioactive metabolites
    From the marine sponge Theonella swinhoei collected off Hachijo-jima Island, we have isolated novel serine protease inhibitors, pseudotheonamides, which were related to the known cyclotheonamides with different modes of cyclization. From the sponge Stelletta globostellatta were isolated three novel isomalabaricane triterpenes which induce morphological changes in rat 3Y1 fibroblasts. We also examined the structure-activity relationships among these triterpenes. Seven 3-alkylpyridine alkaloid were isolated from sponges of genera Xestospongia and Amphimedon. Novel calyculin derivatives were isolated from Discodermia calyx. Chemically modified derivatives and novel natural calyculins were examined for their effects against protein phosphatases and some structure-activity relationships were established.
    2. Studies on the modes of action of bioactive marine natural products
    By using a sponge-derived alkaloid xestospongin C which inhibits the binding of IP3 to its receptor, the involvement of Ca ions in the contraction of guinea-pig papillary muscle.

  • Discovery of Highly Selective Antifungal Agents from Marine Invertebrates

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    1998
    -
    1999
     

    MATSUNAGA Shigeki, NAKAO Yoichi, FUSETANI Nobuhiro

     View Summary

    From Japanese marine invertebrates, following novel compounds have been isolates as a result of bioassay-guided fractionation monitoring antifungal activity.
    1. New mycalolides
    From Mycale sp. collected in the Gulf of Gokasho, three novel mycalolides have been isolated as antifungal constituents. They were assigned as 30-hydroxymycalolide A, 32-hydromycalolide A, and 38-hydroxymycaloude B on the basis of spectral data.
    2. Acanthosterol sulfates
    From Acanthodendrilla sp. collected in the Gulf of Gokasho 10 highly hydroxylated sterol sulfates have been isolated. Their structures were determined by interpretation of spectral data.
    3. Rubrosides and aurantosides
    Eleven compounds that have a tetramic acid and a conjugated polyene chromophore, termed rubrosides and aurantosides, have been isolated from the Siliquariaspongeia japnica collected off Hachijo Island. Their structures were assigned on the basis of spectral data and chemical degradation.

  • 海洋無脊椎動物からの抗トロンビンリード化合物の探索

    日本学術振興会  科学研究費助成事業 奨励研究(A)

    Project Year :

    1998
    -
    1999
     

    中尾 洋一

     View Summary

    伊豆半島,伊豆諸島,九州沿岸,および南西諸島などで採集した海洋無脊椎動物サンプル計342検体(海綿239,腔腸78,原索18,および外肛7検体)について,動物試料20gをアルコールおよびアセトンで抽出し,水とクロロフォルムで二層分配して,水溶性および脂溶性画分を調製した.それぞれの画分について,トロンビンに対する阻害活性スクリーニングを行ったところ,海綿46,腔腸12,原索4,および外肛動物1検体に活性を認めた.
    つぎに,活性の認められた八丈島産海綿Theonella swinhoeiから活性物質の単離を試み,6つの新規活性物質pseudotheonamideA_1,A_2,B_2,C,D,およびdihydrocyclotheonamideAを得た.すなわち海綿サンプルをエタノールで抽出し濃縮後,水とエーテルで二層分配した.水層をさらにブタノールで抽出し,得られたブタノール層を減圧濃縮し,ゲルろ過,ODSフラッシュクロマトグラフィーで順次分画し,最終的に逆相のHPLCを用いて活性物質を精製した.これらの化合物の構造は,MS,NMRなどのスペクトル解析,アミノ酸分析,および化学変換により,絶対立体化学も含めて決定することができた.
    得られた活性物質はトロンビンに対してIC_<50>0.91〜3.0μMで阻害活性を示した.これらの活性は類縁化合物cyclotheonamideAと比べると1/50以下と弱く,k-Argの構造が活性発現に重要であることを裏付けることができた.
    また,中之島産海綿Theonella aff.mirabilisからトロンビン阻害物質を単離中,cathepsinBに対して阻害活性を示すペプチドtokaramideAをあわせて単離・構造決定した.TokaramideAはcathepsinBに対してIC_<50>29ng/mLの阻害活性を示した.

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Misc

  • Efficient Synthesis of Marine Alkaloid Ageladine A and its Structural Modification for Exploring New Biological Activity

    Takayuki Iwata, Koichi Fukase, Yoichi Nakao, Katsunori Tanaka

    JOURNAL OF SYNTHETIC ORGANIC CHEMISTRY JAPAN   78 ( 1 ) 55 - 63  2020.01

    Book review, literature introduction, etc.  

     View Summary

    Natural products have been regarded as a pivotal source for biological active compounds. In addition, structural modification of natural products has been simultaneously received much attention because these modified natural products would show not only enhanced activity and selectivity, but possibly also an entirely distinct bioactivity from those of the parent natural products. We describe herein a series of study on structual modification of ageladine A for exploring new biological activity. Ageladine A was isolated from the marine sponge Agelas nakamurai as an antiangiogenic compound. Based on total syntheses of this alkalod, its structual modifications were caned out as a part of SAR study, which revealed that a pyridine derivative shows a more potent activity. More recently, we have developed an efficient bio-inspired cascade reaction for the synthesis of ageladine A. The cascade reaction involves an aza-electrocyclization and a novel 2-aminoimidazole formation that is modeled after a post-translational modification of arginine residue in protein. Employing the cascade reaction, ageladine A and its N1-substituted analogues were efficiently synthesized in one-pot fashion from anilines or guanidines. In addition, it was found that some analogs showed significant activity on modulating neuronal differentiation. Namely, these analogs selectively activate or inhibit the differentiation of neural stem cells to neurons, while negligible in astrocyte differentiation. The series of study represent a successful case in altering native bioactivity of natural products by structural modification.

  • 海洋微生物共生系を支えるメカニズムの理解にむけた海洋天然化合物プロファイルの解析

    生物工学   97 ( 9 ) 563 - 567  2019  [Invited]

    Authorship:Last author, Corresponding author

  • 時計遺伝子Per2の発現周期やマウス行動周期に対する遠志の影響について

    原口敦嗣, 中村文彬, 阿部真太郎, 中尾洋一, 柴田重信

    日本薬学会年会要旨集(CD-ROM)   138th  2018

    J-GLOBAL

  • 機能性物質増強味噌の試醸と増強方法の検討

    今崎眞司, 林田眞二郎, 松田茂樹, 嶋川淳, 中村文彬, 新井大祐, 加藤妙子, 中野京子, 大池昶威, 中尾洋一

    中央味噌研究所研究報告   39   36 - 42  2018  [Invited]

  • 獅子島産海綿Crella(Yvesia)spinulata由来のcathepsin B阻害物質shishicrellastatin類の単離と構造決定

    村山周平, 菊池条, 中尾洋一, 松永茂樹, VAN SOEST Rob

    日本水産学会大会講演要旨集   2009  2009

    J-GLOBAL

  • 式根島産海綿Asteropus simplex由来のcathepsin B阻害活性物質asteropterinの単離と構造決定

    村山周平, 中尾洋一, 松永茂樹, VAN SOEST Rob

    日本水産学会大会講演要旨集   2008  2008

    J-GLOBAL

  • 海綿Crella(Yvesia)spinulata由来のカテプシンB阻害物質の単離と構造決定

    村山周平, 菊池条, 中尾洋一, VAN SOEST Rob W. M., 松永茂樹

    天然有機化合物討論会講演要旨集   50th  2008

    J-GLOBAL

  • Enzyme Inhibitors from Maine Invertebrates

    Nakao, Y, Fusetani, N

    J. Nat. Prod.   70   689 - 710  2007  [Refereed]

    Authorship:Lead author

  • Perplexing distribution of 3-alkylpyridines in haplosclerid sponges

    Becking, L. E, Nakao, Y, de Voogd, N. J, van Soest, R. M. W

    Porifera Research: Biodiversity, Innovation and Sustainability     173 - 178  2007

  • 式根島産海綿Asteropus simplex由来の新規MMP2阻害活性物質asterodole Aの単離と構造決定

    村山周平, 藤田雅紀, 藤田雅紀, 中尾洋一, 松永茂樹, 伏谷伸宏, 伏谷伸宏

    日本水産学会大会講演要旨集   2006  2006

    J-GLOBAL

  • ナノモルへの挑戦−微量構造解析の最先端−小スケール実験へのマススペクトルの応用

    中尾洋一

      41 ( 7 )  2003  [Invited]

    Authorship:Lead author, Last author, Corresponding author

  • 海洋有用物質の最新研究

      27 ( 4 ) 17 - 21  1999  [Invited]

    Authorship:Lead author

    CiNii

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Industrial Property Rights

  • アミノ酸含有海塩及びその製造方法

    Patent

  • アミノ酸含有海塩及びその製造方法

    新井章吾, 重岡敬之, 吉村博, 中尾洋一

    Patent

  • アストロサイト分化促進用組成物

    中尾 洋一, 伏谷 伸宏, 新井 大祐, 川村 緑

    Patent

  • フェルラ酸誘導体含有組成物及びその製造方法

    中村 文彬, 中尾 洋一, 塩田 邦郎, 新井 大祐

    Patent

  • 白血病幹細胞のニッチ形成抑制活性を有する海洋生物由来の抽出物、化合物及び医薬組成物

    中尾洋一, 浅野茂隆, 新井大祐, 加勢友香梨, 下本哲平

    Patent

  • 白血病幹細胞のニッチ形成抑制活性を有する海洋生物由来の化合物及び組成物

    中尾洋一, 浅野茂隆, 新井大祐, 加勢友香梨, 下本哲平

    Patent

  • アストロサイト分化誘導剤及びGFAP発現誘導剤

    中尾洋一, 新井大祐, 大塚悟史, 片岡亮佑, 斎政彦, 内田裕子

    Patent

  • イミダゾピリジンアミン化合物、その製造方法及び用途

    中尾洋一, 大塚悟史, 田中克典

    Patent

  • 高血糖によるエピゲノム異常の抑制のための方法及び組成物

    中尾洋一, 新井大祐, 塩田邦郎, 早川晃司

    Patent

  • イミダゾールピリジン化合物及びその用途

    中尾洋一, 大塚悟史, 田中克典

    Patent

  • 環状デプシペプチドの新規使用方法

    中尾洋一, 前島寛, 土井隆行, 山下潤, 魚崎英毅, 福島弘之

    Patent

  • 抗リーシュマニア化合物及び抗リーシュマニア薬

    秋山将太, 中尾洋一, 松本芳嗣, 後藤康之, 三條場千寿, 長田康孝, 梅原将洋, 木村純二

    Patent

  • 新規環状デプシペプチドおよびその用途

    中尾 洋一, 勝俣 良祐

    Patent

  • Biologically active compounds derived from marine organisms

    NAKAO Yoichi, MACHIDA Koshi, KIRIKAE Teruo

    Patent

  • Novel Cyclic Depsipeptide and Use Thereof

    YAMASHITA Jun, NAKAO Yoichi, KATSUMATA Ryosuke

    Patent

  • 腔腸動物由来抗原虫化合物

    5721186

    中尾洋一, 石上進太郎, 後藤康之, 河津信一郎, 井上昇

    Patent

  • 新規環状デプシペプチドおよびその用途

    山下潤, 中尾洋一, 勝俣良祐

    Patent

  • 腔腸動物由来抗原虫化合物

    中尾洋一, 石上進太郎, 後藤康之, 河津信一郎, 井上昇

    Patent

  • 環状デプシペプチド

    木村純二, 中尾洋一, 梅原将洋

    Patent

  • 環状デプシペプチド

    木村純二, 中尾洋一, 梅原将洋

    Patent

  • Evaluation of Antiangiogenic Activity of Azumamides by the in vitro Vascular Organization Model Using Mouse Induced Pluripotent Stem (iPS) Cells.

    Patent

  • テロメラーゼ阻害活性を有する新規化合物

    廣田洋, 濱田季之, 松永茂樹, 中尾洋一, 伏谷伸宏, 蕨薫

    Patent

  • 血管新生阻害剤

    中尾洋一, 伏谷伸宏, 清木元治, 渋谷正史, 松永茂樹, 上原至雅, 矢守隆夫, 小野真弓

    Patent

  • 海綿由来アズマミド類

    伏谷伸宏, 松永茂樹, 中尾洋一, 吉田悟, 永井浩二, 新堂信昭, 寺田央

    Patent

▼display all

 

Syllabus

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Teaching Experience

  • 教職課程 化学実験

    Waseda University  

  • 生命化学実験

    Waseda University  

  • Current Topics in Chemistry

    Waseda University  

  • 水圏天然物化学演習

    The University of Tokyo  

  • 実践的化学知セミナーA~B

    Waseda University  

  • ケミカルバイオロジー演習A~D

    Waseda University  

  • 水圏天然物化学実験

    The University of Tokyo  

  • 化学・生命化学特別実験

    Waseda University  

  • 理工学基礎実験2B

  • 理工学基礎実験1B

    Waseda University  

  • 理工学基礎実験1A

    Waseda University  

  • 海洋天然物化学

    Mie University  

  • 静岡ライフサイエンスセミナー

    Shizuoka University  

  • 生命産業R&D特論

    University of Tsukuba  

  • 応用生命化学特別講義II

    University of Tsukuba  

  • 海洋応用生命科学特別講義

    Hokkaido University  

  • 生命科学特論

    Aoyama Gakuin University  

  • 生命科学I

    Aoyama Gakuin University  

  • 水圏生命化学実験法

    The University of Tokyo  

  • 水圏天然物化学特論

    The University of Tokyo  

  • 水圏生命科学特論

    The University of Tokyo  

  • 海へのいざない

    Waseda University  

  • ケミカルバイオロジー特論

    早稲田大学  

  • 生命化学D

    早稲田大学  

  • 生命化学C

    早稲田大学  

  • 化学C

    早稲田大学  

▼display all

 

Social Activities

  • NPO法人 日本水中科学協会シンポジウム

    NPO法人 日本水中科学協会 

    2010
    -
    Now

  • ココア主要成分テオブロミンの新たな生理作用

    第24回 チョコレート・ココア国際栄養シンポジウム 

    2019.09
    -
     

  • 味噌に含有する抗ストレス物質の探索及びその成分を強化したみその製法について

    「食」と「健康」ラボ研究会 第2回シンポジウム 

    2019.01
    -
     

  • 味噌成分のクロマチン修飾におよぼす生理活性プロファイルの解析

    みそサイエンス研究会総会 

  • 発酵食品とエピゲノム情報

    第102回 醸造調味食品セミナー、日本醸造協会 

  • 味噌に含まれる抗ストレス有効成分とその生成過程における発酵の意義

    革新的技術創造促進事業(異分野融合共同研究)市民公開シンポジウム 和食を解き明かす!-世界の健康に貢献する日本食の科学的多面的検証- 

▼display all

Academic Activities

  • Waseda-NUS Joint Symposium, Chemical Epigenomics -The fusion of Epigenetics, Stem Cell Biology and Chemical Biology

    Competition, symposium, etc.

    2009
    -
     
  • 2015環太平洋国際化学会議(PACIFICHEM2015)シンポジウム Frontiers in Chromatin Biology and Chemical Epigenetics/Epigenomics

    Competition, symposium, etc.

  • Frontier research and technologies in cultivation, analyzing, and utilization of microbial dark matter

    Competition, symposium, etc.

  • 日本農芸化学会年会シンポジウム 伝統的発酵食品の生理機能の新展開

    Competition, symposium, etc.

  • 第32回海洋生物活性談話会

    Academic society, research group, etc.

Sub-affiliation

  • Faculty of Science and Engineering   Graduate School of Advanced Science and Engineering

  • Affiliated organization   Global Education Center

Research Institute

  • 2022
    -
    2024

    Waseda Research Institute for Science and Engineering   Concurrent Researcher

  • 2022
    -
    2024

    Waseda Center for a Carbon Neutral Society   Concurrent Researcher

Internal Special Research Projects

  • 心筋分化誘導活性メカニズム解析のためのアプラトキシンプローブ開発

    2023   山本佳奈

     View Summary

    2023年度の特定課題研究では、低濃度で心筋分化を強力に誘導する海洋環状デプシペプチドであるアプラトキシンについて、標的タンパク質を同定するためのプローブ合成を試みた。プローブ合成に用いる標的化合物を3部位に分け、そのうち2部位の合成はほぼ完成した。しかしながら最後のポリケチド部位についてはleighton Allylation、Grignard反応、CBS還元、Fmoc-Proとのカップリング反応による4ステップでの合成達成を目指したが、うまく反応が進まなかった。そこで、既存法と類似した合成法も検討したが、鍵反応の一つが再現できないため、研究に若干の遅れが生じている。ブレークスルーのための反応条件のさらなる検討が必要である。

  • 心筋分化誘導物質アプラトキシンのプローブ合成

    2022   山本佳奈

     View Summary

    われわれが非常に強力な心筋分化誘導活性を見出した、海洋環状デプシペプチドApratoxin類について、作用メカニズムの解析を行ってきた。これまでに用いたプローブ分子の構造が適切でなかったため、標的タンパク質の同定および作用メカニズムの解明には至っていないことをふまえ、本課題では直接的に標的タンパク質を同定するためのプローブの構造を再検討して再合成を行うこととした。学内の共同研究(山本佳奈准教授)により、中尾研究室の学生が山本准教授の指導の下で合成研究を行った。この結果、新たなリンカー導入部位を含むプローブ分子について、ペプチド部の合成を達成し、ポリケチド部分の合成も進んでおり、目標とする期間内(1年間)での合成の達成が見えてきた。

  • 心筋分化誘導物質Apratoxin類の作用メカニズム解析

    2021  

     View Summary

    われわれは海洋環状ペプチドApratoxin類に非常に強力な心筋分化誘導活性を見出しており、その作用メカニズムの解析を行ってきた。これまでに標的タンパク質釣り上げ用のプローブを用いた標的タンパク質解析を行い、いくつかの候補タンパク質について組換え体を作成して、化合物との相互作用を解析してきたが、いずれの候補タンパク質においても、非常に積陽作用を明確に説明できる結果が得られていない。そこで、タンパクつり上げ実験と並行して作用メカニズム解析が可能となるshRNAライブラリーによる作用メカニズム解析の準備を進めた。

  • shRNAライブラリースクリーニングによる天然由来化合物の作用機序解明

    2020   新井大祐

     View Summary

    これまでの研究で、CRISPRゲノムワイドスクリーニングの構築に必要なレンチウイルスライブラリー(HumansgRNA library Brunello in lentiCRISPRv2, Mouse sgRNA library Brie inlentiCRISPRv2)をAddgene社より購入した。また、レンチウイルスの作製に必要な市販のパッケージング細胞(293LTV)とパッケージングプラスミド(psPAX2, PMD2.G)も購入し、パッケージング細胞は培養、増殖させ凍結ストックを作製した。

  • ヒストン修飾調節を行う小分子の探索と機能解析

    2019   神平梨絵

     View Summary

    エピジェネティックな遺伝子発現の調節(遺伝子スイッチ)にもかかわり、種々の疾病の原因になると考えられているヒストンタンパク質の化学修飾の異常をコントロールし、正常に戻すような医薬品や機能性食品への応用を目指して、ヒストン修飾を調節する化合物の探索を食品や海産物の有効成分から行った。本度の研究では独自に開発したセルベースのヒストン修飾評価法を用いて、特定のヒストン修飾に変化を引き起こす複数の海洋天然化合物や食品成分を見出し、海外および国内学会で発表するとともに、化粧品開発展において展示を行った。

  • 白血病幹細胞ニッチを標的とした新規抗がん剤の探索

    2016  

     View Summary

    本研究では、白血病幹細胞(LSC)の性質を有する株化細胞であるMB-1細胞によるニッチ形成を簡便にin vitroで再現できるアッセイ系を用いて、白血病幹細胞に効く抗がん剤を探索した。この結果、われわれが独自に保有する海洋生物抽出物ライブラリーから、低濃度でMB-1細胞のニッチ形成を阻害する化合物を見出すことができた。ニッチを形成したMB-1細胞は慢性骨髄性白血病の第1選択薬であるイマチニブに対して薬剤抵抗性を示すことが知られているため、本化合物によるニッチ形成阻害が薬剤抵抗性におよぼす影響を解析したところ、本化合物によってMB-1細胞のイマチニブ感受性が回復することを明らかにできた。

  • 白血病幹細胞ニッチを標的とした新規抗がん剤の探索

    2015   浅野茂隆

     View Summary

    白血病幹細胞(LSC)の性質を有する株化細胞であるMB-1細胞を用いたアッセイ系によって、独自に保有する海洋生物抽出物ライブラリーから白血病幹細胞のニッチ形成を標的とする抗がん剤リード化合物の探索を行った。ここで活性化合物として得られた海洋生物由来天然化合物Aについて、MB-1細胞のCA形成に対する影響を解析したところ、本来はヒト慢性骨髄性白血病の第一選択薬のイマチニブ(IM)に耐性であるMB-1細胞のIM感受性が回復することが示唆された。以上のように、白血病幹細胞のニッチ形成を標的とする新たな抗がん剤リード化合物を得ることができた。

  • 多剤耐性緑膿菌に対する抗菌剤の作用メカニズム解析

    2014   切替照夫

     View Summary

    本研究では、海洋生物エキスから得られた、多剤耐性緑膿菌に対する抗菌化合物Aの作用メカニズムを明らかにして、有効な薬剤開発につなげるための基礎的知見を得ることを目的として研究を行った。具体的には単離した抗菌化合物Aに対する耐性株の作成を行い、耐性株のゲノム配列を次世代シーケンサーで解析し、変異点をみつけることによって見出した抗菌化合物Aの標的遺伝子を探索した。この結果、明らかとなった標的遺伝子候補について、大腸菌による組み換え体の発現を試み、抗菌化合物Aに対する耐性株由来の変異酵素については発現を確認している。現在は化合物A感受性株由来の組み換え酵素の発現条件の検討中である。

  • 海洋生物由来抗原虫活性化合物のケミカルバイオロジー研究

    2013  

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    リーシュマニア症はサシチョウバエによって媒介される節足動物媒介性人獣共通感染症であり、WHOによって対策の最も困難な熱帯感染症(カテゴリーI)として位置づけられ、重点対策10大熱帯感染症の一つにも指定されている。そこで本研究ではユニークな構造と生物活性を有する化合物の宝庫である海洋生物を探索源として、抗リーシュマニア薬開発につながるような化合物の探索を行うこととした。本研究では緑色蛍光タンパク質(GFP)導入原虫を用いた迅速な抗原虫活性評価法を用いて、幅広く海洋無脊椎動物抽出物について研究分担者後藤がスクリーニングを行い、抗リーシュマニア活性化合物の探索を行った。スクリーニングにより活性が認められた海綿サンプルについて、活性化合物の単離を試みた結果、ミクロネシア産海綿に含まれる抗リーシュマニア活性化合物を単離・同定することができた(学会発表1)。一方、以前の研究で見出された強い抗リーシュマニア活性を有する化合物cristaxenicin Aについて、in vitroの活性評価および作用メカニズム解析のためのプローブ開発に向けて、研究分担者の細川が全合成研究を継続している。また、これまでに得られている抗リーシュマニア活性化合物ciliatamideについて、研究分担者木村がciliatamideを構成する環状アミノ酸部(c-Lys)、アミノ酸部(Me-Phe)、脂肪酸部を入れ替えた類縁体ライブラリーを合成し、活性評価を行った。この結果、c-Lysは環状アミンでも代替がきくこと、Me-Pheの他のアミノ酸への置換は活性に大きく影響すること、脂肪酸部分は炭素数が10程度のものが望ましいことが明らかとなった(学会発表2)。本化合物は比較的単純なユニットで活性を保持した誘導体を簡便に合成できるため、今後のin vitro活性評価や作用メカニズム解析のプローブ分子への転用が期待できる。

  • 多剤耐性緑膿菌に対する抗菌剤の探索

    2013  

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    2010以来、多剤耐性菌による院内感染が国内でも大きな問題となりつつあり、その対策が急務となっている。2010年に問題になった院内感染ではアシネトバクターが大きく取り上げられていたが、実際には多剤耐性緑膿菌(Multi-Drug Resistant Pseudomonas aeruginosa: MDRP)も検出されており、院内感染の現場においてはむしろこちらのほうが大きな問題となっている。緑膿菌は、生来多くの抗菌剤に耐性であるため、セフタジディムやイミペネム等のβ-ラクタム剤やアミカシンやトブラマイシン等のアミノグリコシド剤など、ごく限られた中から選んだ薬剤を治療に用いられているが、1980年代初頭から、これらのカルバペネム系、フルオロキノロン系、アミノグリコシド系に対しても、3系統の抗菌剤全てに耐性を示す多剤耐性緑膿菌が出現しているため、これらの耐性菌に対する対策が急務となっている。そこで本研究では、多剤耐性緑膿菌に対して有効な抗菌剤開発のリード化合物を探索するため、表現型指向性アッセイである多剤耐性菌を用いた抗菌活性試験と、薬剤耐性の原因となる酵素のAACおよびMBLに対する酵素阻害活性を標的指向型アッセイとして組み合わせ、多剤耐性を克服できるような新規抗菌剤を見出すことを目的とした。本研究は多剤耐性菌と戦う最前線の現場である国立国際医療研究センターとの密接なコラボレーションによって行い、以下に述べる成果をあげることができた。当研究室が保有する海洋無脊椎動物抽出物1816検体についてMIC法による抗菌活性を評価したところ、7検体に抗菌活性が確認された。特に顕著な活性を示した鹿児島県産Agelas属海綿から活性本体の探索を試みた。本海綿をメタノールで抽出し、抽出液を濃縮後、水とクロロホルムにより二層分配し、水層はさらにn-ブタノールで抽出した。クロロホルム層とn-ブタノール層を合一して濃縮後、ヘキサン層と90%メタノール層に二層分配した。その後、活性を示した90%メタノール可溶性画分をODSカラムクロマトグラフィーによって分画し、6つの画分を得た。最も強い抗菌活性を示し、かつサンプル量が豊富であった70%メタノール可溶性画分を逆相HPLCに付し、活性主成分として大腸菌、および枯草菌に対する抗菌活性物質として知られるoroidin、およびその類縁体を同定した(MIC 6.25~50 &micro;g/mL)。得られた化合物の多剤耐性緑膿菌に対する抗菌活性、およびHeLa細胞、P388細胞に対する細胞毒性を評価した。これらの化合物の共通の特徴はブロモピロールと2-アミノイミダゾールを含むことであった。oroidinのダイマーを基本骨格としているageliferin、bromoageliferin、dibromoageliferin、mauritiamineはoroidinを上回る抗菌活性を示し、2-アミノイミダゾールの部分構造を含まない4,5-dibromopyrroll-2-carboxylic acidは抗菌活性を示さなかった。特に、ageliferinは最も強い抗菌活性を示し、最も弱い細胞毒性が確認された。そこで本化合物の作用機序を研究するため、本化合物に対する耐性株を作製した。耐性株に対する抗菌活性はNCGM2.S1株のそれと比較して8倍以上のMIC値を示した。次世代シーケンサーによって本耐性菌株のゲノムを解析し、変異点から標的タンパク質の絞り込みを行って作用メカニズムの解析を行っている。

  • アジア・太平洋海域における有用海洋生物資源調査・2

    2012  

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    2012年度は国内(南西諸島:奄美大島、徳之島、中之島、大島新曽根、竹島、伊豆諸島:式根島)で海綿、腔腸、原索動物などの海洋無脊椎動物を中心として139検体の海洋生物サンプルを採集した。採集したサンプルに対して、アルコール抽出エキスから調製した脂溶性および水溶性成分のライブラリーを作成した。同時に共生微生物のゲノム遺伝子採取用にサンプル小片をエタノール中冷凍保存し、遺伝子ライブラリーとした。また、生理活性化合物を作っている共生微生物の単離・同定・培養法確立のため、式根島産の海綿 Discodermia calyxを採集し、実験室内で長期飼育を行い、飼育サンプルから共生微生物を取り出して、メッシュによる分画とセルソーターによる共生微生物の分離を試みた。得られた微生物画分に関して、産業総合科学研究所(木村信忠主任研究員)およびNITE(山副敦司研究職員)との共同研究によって次世代シーケンサーを用いたメタゲノム解析を実施し、主要な共生微生物の同定およびゲノム遺伝子解析を行った。一方、イタリア・ナポリ大学のグループ(M. V. D’Auria教授およびA. Zampella教授)と新たにTheonella 属の海綿に含まれる、ステロイド誘導体成分の構造-活性相関研究に関する共同研究を開始した。まず天然の主成分であるセオネラステロールを大量に単離して、天然化合物をメチル化・アセチル化・酸化・還元・脱水などの化学的操作によって各種誘導体に導いた。導いた各誘導体をつかって、胆汁鬱滞の治療薬標的として期待されるFXRアゴニスト活性を調べ、さらにドッキングモデル解析を行うことで、構造-活性相関を明らかにした。(論文1)本共同研究が基盤となり、2013年度の基盤研究(B)海外学術調査は、イタリアのグループとの共同研究の内容「海洋微生物の共生-進化系理解のための日-欧連携フィールドワーク」で研究助成申請を行い、採択された。

  • iPS細胞へのリプログラミング活性を有する海洋天然化合物に関する研究

    2011  

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    本研究では、遺伝子の改変なく細胞をリプログラミングしてiPS細胞をへと導かせることができるような手法の開発を目指して、iPS細胞誘導に必要な4遺伝子の機能を代替できるような化合物を海洋生物から探索することを目的とした。探索源となる海洋生物はすでに所有する約1500検体に加えて、新たに約250サンプルを日本各地およびインドネシアにて採集した。採集したサンプルから調製した抽出物ライブラリーに対して、2通りの方法で活性化合物の探索を行った。1.まず、京都大学山中らのプロトコールに従い、Nanog-GFP-puroマウスから取り出した繊維芽細胞に対して、山中カクテルと呼ばれる4因子(Oct4、Klf4、Sox2、cMyc)の遺伝子導入を行うと、GFPを発現し緑色に光るiPS細胞を誘導できる。本研究では山中4因子から1因子ずつを取り除き、3因子を導入した状態の細胞に対してサンプルを投与して、取り除いた1因子の機能を代替してiPS細胞を誘導しうるような活性を有するサンプルを探索する。これまでに、Oct4を除いた3遺伝子を導入した細胞に対するiPS細胞誘導活性スクリーニングを行い、複数のサンプルでGFPを発現したコロニーを見出しているので、現在これらのサンプルから活性本体の精製を行っている。2.次に、マウスES細胞を用いて、ES細胞の未分化状態を維持する活性を有する化合物の探索を行った。ES細胞の未分化状態を維持する活性を有する化合物は、iPS細胞の誘導時に補助的な機能が期待されるばかりでなく、作用メカニズムの解析によって細胞の分化制御機構をあきらかにできれば、より効率的なiPS細胞の誘導法の開発にとって有用な知見となりうる。これまでの研究で、50pg/mLという極めて低濃度でES細胞の未分化状態を維持できる化合物を見出すことに成功した。本化合物投与下で培養したES細胞をヌードマウスに移植したところ、テラトーマが形成され、三胚葉すべての組織に分化していることが確認されたため、本化合物によってES細胞の未分化性が維持されていることがわかった。現在、その作用メカニズムの解析を行っている。

  • 動的平衡としての海洋微生物共生系のケミカルバイオロジー

    2010  

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    カリキュリンを含む海綿Discodermia calyxの新たな生息地を首都圏近郊に発見することができたので生息域の違いによる含有化合物の違いを検討している。また、人工飼育中の海綿から随時共生微生物を取り出して、カリキュリンの生産微生物を含む微生物群について、抽出したゲノム遺伝子を用いてメタゲノム解析の準備を行っている。メタゲノム解析が終了次第、PKSおよびNRPS遺伝子配列をもとにカリキュリン生合成遺伝子の遺伝子配列の解析を行う。一方、新たな生合成遺伝子探索のターゲットとして、抗がん剤エクテナサイジンと同じ骨格を有するレニエラマイシンを選び、本化合物を含有する海綿の採集および人工飼育を検討した。この結果、本海綿はD.calyxと異なり、約1カ月程度しか飼育下では生存できないことが明らかとなったため、採集方法および飼育条件の検討を行っている。なお、本海綿から共生微生物を取り出し、化合物の含有量をもとにレニエラマイシン産生微生物の絞り込みを行い、得られた候補微生物に関してメタゲノム解析の準備を進めている。

  • 医薬品・生化学試薬としての海洋天然化合物の再探索

    2010  

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    日本およびアジア太平洋各地で採集した海洋無脊椎動物からスクリーニング用サンプルの調製を行った。調製したスクリーニングサンプルに対して、抗がん、抗原虫、幹細胞分化制御活性などのスクリーニングを行い、ヒットサンプルを浮かび上がらせた。抗原虫活性を示した鹿児島県の海底火山で採集した腔腸動物ヤギ類から、クリスタキセニシンと命名した新規化合物を活性本体として単離し、その構造を明らかにした。本化合物は低細胞毒性でリーシュマニア原虫に対する選択的抗原虫活性を示したため、国際特許出願を行った。また、宮城県産のホヤから、マウスES細胞に対する未分化維持活性を有する新規化合物を単離、構造決定した。本化合物は1ng/mL以下のごく低濃度でマウスES細胞の未分化状態を維持するとともに、本化合物投与下で維持されたES細胞をヌードマウスに皮下移植するとテラトーマを形成し、三胚葉すべての組織への分化が認められたことから、本化合物で維持されたES細胞は万能性も維持されていることを明らかにできた。以上の結果から、本化合物について米国仮出願申請を行った。

  • カリキュリン生産に関わる海洋微生物共生系のケミカルバイオロジー

    2009  

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    海綿に代表される海洋無脊椎動物は、陸上の生物が作り出す化合物とは構造が大きく異なるユニークな化合物の宝庫であることが知られ、医薬品素材の探索源としても有望視されている。一方、多くの海洋天然化合物が海洋無脊椎動物に共生している微生物によって生合成されていると考えられている。そこで本研究では、代表的な海洋天然化合物として知られているカリキュリンを含む海綿Discodermia calyxをモデルに選び、海綿という海洋微生物共生系における化合物の産生機構の理解につながる知見を得ることを目的として研究を行っている。まず、海綿D. calyxを採集し、研究室内で飼育することにより、いつでも共生微生物が存在した健康的な状態の海綿を実験に使えるような飼育条件を検討した。これまでの研究において、室温で飼育すると1週間程度しか海綿が健康な状態で生存できないこと、低温(4度)で飼育すると相当期間飼育が可能なことが分かっていたため、海綿の健康的な状態を保つためには温度が非常に重要なファクターであると考え、水温30度、25度、20度にて飼育実験を行ってみたところ、20度で飼育すれば良好な健康状態を保ったまま飼育ができることを明らかにできた。一方、カリキュリンを生合成していると考えられる微生物は我々の研究結果から16S rDNAの配列解析から、最近になって初めて存在が明らかになったOD1(OP11-derived 1, OP= Obsidian Pool)という新しい門に属する培養不可能な微生物に近いものであることが示唆されている。そこで、30度と20度で飼育した海綿から経時的に体の一部を切り取り、そこに含まれる微生物相の変化を遺伝解析によって行い、カリキュリン含量の変化とリンクさせて比較解析することで、カリキュリン生合成微生物の特定を行っている。

  • 新規エピジェネティック治療薬の探索研究

    2008  

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    『エピジェネティクス』とは『世代を超えて継承され得る、塩基配列を伴わない遺伝子発現の変化』のことであり、おもにDNAのメチル化とヒストンの修飾によって多くの遺伝子の発現が制御されうることが知られている。がん治療では、DNAメチル化酵素(Dnmt)阻害剤やヒストン脱アセチル化酵素(HDAC)阻害剤を同時投与することによる相乗効果でがん抑制遺伝子の発現を高いレベルで誘導可能になると期待され、欧米を中心にDnmt阻害剤やHDAC阻害剤をがん治療に応用しようというエピジェネティック治療が推し進められており、現在、Dnmt阻害剤の5-Aza-CdRおよび5-Aza-CRが骨髄異型性症候群の治療薬として、HDAC阻害剤のSAHAが皮膚T細胞リンパ腫の治療薬としてFDAに承認され、治療効果をあげている一方、研究代表者はこれまで海洋無脊椎動物を対象として、さまざまな酵素に対する阻害剤の探索研究を行い、鹿児島県産の海綿からアズマミドA-E(AzA-E)と命名した5種類の新規HDAC阻害剤を見出している。以上のような背景下、本研究では、酵素阻害剤の宝庫である海洋無脊椎動物を探索対象として新たなDnmt阻害剤の探索を目的として研究を行った。まず、1型のDNAメチル化酵素であるDnmt1によるDNAメチル化阻害活性を調べるアッセイ系を確立した。ついで本アッセイ系を用いたスクリーニングを、日本近海で採集した海洋無脊椎動物の抽出物ライブラリー約400サンプルについて行い、約50サンプルに活性を認めた。今後はスクリーニングを継続するとともに、ヒットサンプルから活性化合物の単離・構造決定を行う予定である。

  • 新規抗リーシュマニア剤の探索研究

    2008  

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    本研究はリーシュマニア症に苦しむ発展途上国の人々の窮状を救うべく、医薬品素材となる新規化合物の宝庫である海洋無脊椎動物から、抗リーシュマニア原虫作用を示す化合物を探索することを目的とした。まず、GFP導入原虫(LaEGFP)を用いた、細胞内アマスティゴートに対する迅速な薬剤試験方法を用いて海洋無脊椎動物抽出物から、抗原虫活性を有する化合物の探索、同定を目的とし、研究代表者が日本近海で採集した海洋生物からスクリーニングサンプル作り、活性化合物の単離・構造決定、および化合物の誘導体化は研究代表者が行い、抗原虫活性試験は研究協力者が行った。スクリーニングサンプル:採集した海洋無脊椎動物をアルコールで抽出し、濃縮後水とクロロホルムで二層分配し、それぞれの層を濃縮してスクリーニングサンプルとした。スクリーニング:以上のように調整したスクリーニングサンプル計1360を抗原虫活性スクリーニングに付した。方法としては、96穴プレートにマウスマクロファージ細胞J774を1x106個まき、その後2x107個の原虫で感染させた。16時間後に非感染原虫をPBS洗浄により除いた後、培地によって希釈された化合物を加えて48時間培養した。その後蛍光プレートリーダーで各ウェルの蛍光強度を測定して、その強度により原虫数および薬剤の抗原虫活性の算出を行った。アンフォテリシンBを用いた試験においては薬剤存在下において原虫由来GFPの減衰がみられ、蛍光強度から算出されたIC50の値は、従来の方法(顕微鏡観察による原虫数の計測)を用いて求められた値と極めて近い値を示し、この方法の有用性が確認されている。活性化合物の単離・構造決定:スクリーニングで浮かび上がった鹿児島県奄美大島産の海綿サンプルから抗原虫活性を指標として3種の活性化合物の単離を行い、単離した化合物の構造をはMS、NMRなどのスペクトル分析により既知のaaptamine (1)、isoaaptamine (2)、および9-demethylaaptamine (3)であると同定した。化合物の活性評価:以上の得られた化合物1-3について抗原虫活性を調べたところ、LaEGFPに対してそれぞれIC50 2.3、0.9、および0.5ug/mLの抗原虫活性を示した。また、鹿児島県産の海綿から単離された新規化合物glacilioether Bに抗リーシュマニア原虫活性を見出すとともに(68%阻害 10ug/mL)、別種の海綿から新規抗リーシュマニア化合物ciliatamides AおよびBを見出した(IC50 10ug/mL)。

  • 海洋生物由来血管新生阻害剤の探索

    2007  

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    ユニークな構造と強烈な生物活性を有する化合物を豊富に含むことが知られる海洋無脊椎動物を探索源として、がんの増殖・転移に深くかかわることが知られる血管新生を阻害する新規化合物の探索研究を行った。1.まず、日本各地(鹿児島から青森までの5か所)において海洋無脊椎動物サンプルの採集を行い、約200検体のサンプルを採集した。2.次いで、これらのサンプルをアルコールで抽出し、水とクロロホルムで二層分配後、濃縮してそれぞれの層を水溶性画分および脂溶性画分とした。これらの各分をさらにC18のカラムを用いた固相抽出法にてそれぞれ6つに分画し、最終的に約2000のスクリーニング用サンプルを調製した。3.調製したスクリーニングサンプルについては現在血管新生阻害活性スクリーニングを開始している。4.血管の管腔構造構築を阻害する活性を有する鹿児島県産Sinularia属の軟サンゴから、活性本体として、新規オキシリピンを単離構造決定した(投稿中の論文参照)。5.また、鹿児島県産のMicale属の海綿から単離していた血管新生阻害作用を有するヒストン脱アセチル化酵素(HDAC)阻害剤azumamideについて、京都大学山中らによって開発された人工多能性(iPS)細胞を用いた血管再構築系によって抗血管新生作用を調べ、顕著な血管新生阻害活性を見出し報告した(世界最初のiPS細胞を用いたケミカルバイオロジー研究の論文)。以上の研究を通して海洋無脊椎動物の血管新生阻害剤探索源としての有望性を示すことができたと同時に、現在世界中で極めて熾烈な研究競争が行われているiPS細胞の研究においても世界に先駆ける研究成果を挙げることができた。

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