Updated on 2022/09/25

写真a

 
FUTAMURA, Yushi
 
Affiliation
Faculty of Science and Engineering, Waseda Research Institute for Science and Engineering
Job title
Junior Researcher(Assistant Professor)

Education

  • 2002.04
    -
    2008.03

    Keio University  

  • 1998.04
    -
    2002.03

    Keio University   Faculty of Science and Technology  

Degree

  • 慶應義塾大学   博士(理学)

Research Experience

  • 2022.04
    -
    Now

    Waseda University   Research Institute for Science and Engineering

  • 2015.04
    -
    2022.03

    理化学研究所   環境資源科学研究センター   研究員

  • 2013.08
    -
    2015.03

    公益財団法人肝炎ウイルス研究財団   リサーチレジデント

  • 2011.10
    -
    2013.07

    理化学研究所   長田抗生物質研究室   特別研究員

  • 2008.10
    -
    2011.09

    理化学研究所   長田抗生物質研究室   基礎科学特別研究員

  • 2008.04
    -
    2008.09

    理化学研究所   長田抗生物質研究室   協力研究員

▼display all

Professional Memberships

  •  
     
     

    JAPANESE SOCIETY FOR CHEMICAL BIOLOGY

  •  
     
     

    THE JAPANESE ASSOCIATION FOR MOLECULAR TARGET THERAPY OF CANCER

  •  
     
     

    THE JAPANESE CANCER ASSOCIATION

  •  
     
     

    JAPAN SOCIETY FOR BIOSCIENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY

 

Research Areas

  • Applied microbiology

  • Chemical biology

Research Interests

  • 表現型スクリーニング

  • 天然物

  • Chemical Biology

Papers

  • Phenylboronic Ester-Activated Aryl Iodide-Selective Buchwald−Hartwig-Type Amination toward Bioactivity Assay

    Raghu N. Dhital, Abhijit Sen, Hao Hu, Rikako Ishii, Takuma Sato, Yoko Yashiroda, Hiromi Kimura, Charles Boone, Minoru Yoshida, Yushi Futamura, Hiroyuki Hirano, Hiroyuki Osada, Daisuke Hashizume, Yasuhiro Uozumi, Yoichi M.A. Yamada

    ACS Omega    2022.07  [Refereed]

    DOI

  • New antimalarials identified by a cell-based phenotypic approach: Structure-activity relationships of 2,3,4,9-tetrahydro-1H-β-carboline derivatives possessing a 2-((coumarin-5-yl)oxy)alkanoyl moiety.

    Nobuo Cho, Ko Kikuzato, Yushi Futamura, Takeshi Shimizu, Hiroki Hayase, Kikuko Kamisaka, Daisuke Takaya, Hitomi Yuki, Teruki Honma, Mamoru Niikura, Fumie Kobayashi, Nobumoto Watanabe, Hiroyuki Osada, Hiroo Koyama

    Bioorganic & medicinal chemistry   66   116830 - 116830  2022.05  [International journal]

     View Summary

    The identification, structure-activity relationships (SARs), and biological effects of new antimalarials consisting of a 2,3,4,9-tetrahydro-1H-β-carboline core, a coumarin ring, and an oxyalkanoyl linker are described. A cell-based phenotypic approach was employed in this search for novel antimalarial drugs with unique modes of action. Our screening campaign of the RIKEN compound library succeeded in the identification of the known tetrahydro-β-carboline derivative (4e) as a hit compound showing significant in vitro activity. SAR studies on this chemical series led to the discovery of compound 4h having a (R)-methyl group on the oxyacetyl linker with potent inhibition of parasite growth (IC50 = 2.0 nM). Compound 4h was also found to exhibit significant in vivo antimalarial effects in mouse models. Furthermore, molecular modeling studies on 4e, 4h, and its diastereomer (4j) suggested that the (R)-methyl group of 4h forces the preferential adoption of a specific conformer which is considered to be an active conformer.

    DOI PubMed

  • Isolation of new lucilactaene derivatives from P450 monooxygenase and aldehyde dehydrogenase knockout Fusarium sp. RK97-94 strains and their biological activities.

    Islam A Abdelhakim, Takayuki Motoyama, Toshihiko Nogawa, Fauze Bin Mahmud, Yushi Futamura, Shunji Takahashi, Hiroyuki Osada

    The Journal of antibiotics    2022.04  [International journal]

     View Summary

    Fusarium sp. RK97-94 is a producer of potent antimalarial compounds such as lucilactaene and its derivatives. The biosynthetic gene cluster of lucilactaene was identified but only a knockout mutant of methyltransferase (luc1) was reported in previous papers. Herein, we report on isolation and identification of prelucilactaene G (1), and prelucilactaene H (2) from the aldehyde dehydrogenase knockout strain (∆luc3) culture broth, as well as prelucilactaene A (3), prelucilactaene B (4), and two isomeric mixtures of prelucilactaene E (5) and prelucilactaene F (6), from the P450 monooxygenase knockout strain (∆luc2) culture broth. Our data, unlike the previous ones, suggest the involvement of the aldehyde dehydrogenase (Luc3) in lucilactaene biosynthesis, and support the involvement of the P450 monooxygenase (Luc2) in C-20 hydroxylation rather than C-13-C-14 epoxidation or C-15 hydroxylation. Isolated compounds displayed moderate to strong antimalarial activities, and the structure-activity relationship of lucilactaene derivatives was examined.

    DOI PubMed

  • Establishment of a Monoclonal Antibody against Human NTCP That Blocks Hepatitis B Virus Infection.

    Toshitada Takemori, Akiko Sugimoto-Ishige, Hironori Nishitsuji, Yushi Futamura, Michishige Harada, Tomomi Kimura-Someya, Takehisa Matsumoto, Teruki Honma, Miho Tanaka, Masami Yaguchi, Kyoichi Isono, Haruhiko Koseki, Hiroyuki Osada, Daiki Miki, Takashi Saito, Takashi Tanaka, Takehiro Fukami, Toshio Goto, Mikako Shirouzu, Kunitada Shimotohno, Kazuaki Chayama

    Journal of virology   96 ( 5 ) e0168621  2022.03  [International journal]

     View Summary

    Hepatitis B virus (HBV) infects 240 million people worldwide. Current therapy profoundly suppresses HBV replication but requires long-term maintenance therapy. Therefore, there is still a medical need for an efficient HBV cure. HBV enters host cells by binding via the preS1 domain of the viral L protein to the Na+/taurocholate cotransporting polypeptide (NTCP). Thus, NTCP should be a key target for the development of anti-HBV therapeutics. Indeed, myrcludex B, a synthetic form of the myristoylated preS1 peptide, effectively reduces HBV/hepatitis D virus (HDV) infection and has been approved as Hepcludex in Europe for the treatment of patients with chronic HDV infection. We established a monoclonal antibody (MAb), N6HB426-20, that recognizes the extracellular domain of human NTCP and blocks HBV entry in vitro into human liver cells but has much less of an inhibitory effect on bile acid uptake. In vivo, administration of the N6HB426-20 MAb prevented HBV viremia for an extended period of time after HBV inoculation in a mouse model system without strongly inhibiting bile acid absorption. Among the extracellular loops (ECLs) of NTCP, regions of amino acids (aa) 84 to 87 in ECL1 and aa 157 to 165 near ECL2 of transmembrane domain 5 are critically important for HBV/HDV infection. Epitope mapping and the three-dimensional (3D) model of the NTCP structure suggested that the N6HB426-20 MAb may recognize aa 276/277 at the tip of ECL4 and interfere with binding of HBV to the region from aa 84 to 87. In summary, we identified an in vivo neutralizing NTCP-targeting antibody capable of preventing HBV infection. Further improvements in efficacy of this drug will pave the way for its clinical applications. IMPORTANCE A number of entry inhibitors are being developed to enhance the treatment of HBV patients with oral nucleoside/nucleotide analogues (NA). To amplify the effectiveness of NA therapy, several efforts have been made to develop therapeutic MAbs with neutralizing activity against HBs antigens. However, the neutralizing effect of these MAbs may be muted by a large excess of HBsAg-positive noninfectious particles in the blood of infected patients. The advantage of NTCP-targeted HBV entry inhibitors is that they remain effective regardless of viral genotype, viral mutations, and the presence of subviral particles. Although N6HB426-20 requires a higher dose than myrcludex to obtain equivalent suppression of HBV in a model mouse system, it maintained the inhibitory effect for a long time postadministration in proportion to the half-life of an IgG MAb. We believe that further improvements will make this antibody a promising treatment option for patients with chronic hepatitis B.

    DOI PubMed

  • Dihydrolucilactaene, a Potent Antimalarial Compound from Fusarium sp. RK97-94.

    Islam A Abdelhakim, Fauze Bin Mahmud, Takayuki Motoyama, Yushi Futamura, Shunji Takahashi, Hiroyuki Osada

    Journal of natural products   85 ( 1 ) 63 - 69  2022.01  [International journal]

     View Summary

    A recently discovered secondary metabolism regulator, NPD938, was used to alter the secondary metabolite profile in Fusarium sp. RK97-94. Three lucilactaene analogues were detected via UPLC-ESI-MS analysis in NPD938-treated culture. The three metabolites were successfully purified and identified as dihydroNG391 (1), dihydrolucilactaene (2), and 13α-hydroxylucilactaene (3) via extensive spectroscopic analyses. DihydroNG391 (1) exhibited weak in vitro antimalarial activity (IC50 = 62 μM). In contrast, dihydrolucilactaene (2) and 13α-hydroxylucilactaene (3) showed very potent antimalarial activity (IC50 = 0.0015 and 0.68 μM, respectively). These findings provide insight into the structure-activity relationship of lucilactaene and its analogues as antimalarial lead compounds.

    DOI PubMed

  • β-Methyltryptamine Provoking the Crucial Role of Strictosidine Synthase Tyr151-OH for Its Stereoselective Pictet-Spengler Reactions to Tryptoline-type Alkaloids.

    Haicheng Liu, Santosh Panjikar, Xiang Sheng, Yushi Futamura, Chenghua Zhang, Nana Shao, Hiroyuki Osada, Hongbin Zou

    ACS chemical biology   17 ( 1 ) 187 - 197  2022.01  [International journal]

     View Summary

    Strictosidine synthase (STR), the gate enzyme for monoterpenoid indole alkaloid biosynthesis, catalyzes the Pictet-Spengler reaction (PSR) of various tryptamine derivatives with secologanin assisted by "indole sandwich" stabilization. Continuous exploration with β-methyltryptamine (IPA) stereoselectively delivered the C6-methylstrictosidines and C6-methylvincosides by enzymatic and nonenzymatic PSR, respectively. Unexpectedly, the first "nonindole sandwich" binding mode was witnessed by the X-ray structures of STR1-ligand complexes. Site-directed mutagenesis revealed the critical cryptic role of the hydroxyl group of Tyr151 in IPA biotransformation. Further computational calculations demonstrated the adjustable IPA position in STR1 upon the binding of secologanin, and Tyr151-OH facilitates the productive PSR binding mode via an advantageous hydrogen-bond network. Further chemo-enzymatic manipulation of C6-methylvincosides successfully resulted in the discovered antimalarial framework (IC50 = 0.92 μM).

    DOI PubMed

  • 2-Methylthio-N7-methyl-cis-zeatin, a new antimalarial natural product isolated from a Streptomyces culture.

    Julius Adam V Lopez, Toshihiko Nogawa, Kazuko Yoshida, Yushi Futamura, Hiroyuki Osada

    Bioscience, biotechnology, and biochemistry   86 ( 1 ) 31 - 36  2021.12  [International journal]

     View Summary

    2-Methylthio-N7-methyl-cis-zeatin (1) was isolated from the culture broth of Streptomyces sp. 80H647 along with 2 known purine derivatives, 5'-methylthioinosine (2) and AT-265 (dealanylascamycin, 3). The structure elucidation of compound 1 was accomplished by high-resolution mass spectrometry (HRMS) and nuclear magnetic resonance (NMR) analyses. It inhibited the growth of Plasmodium falciparum 3D7 with a GI50 of 2.4 µm and had no effect on the growth of Arabidopsis at 2 µm. This is the first report of an N7-methylated zeatin-type natural product from Streptomyces and as an antimalarial compound.

    DOI PubMed

  • Privileged Biorenewable Secologanin-Based Diversity-Oriented Synthesis for Pseudo-Natural Alkaloids: Uncovering Novel Neuroprotective and Antimalarial Frameworks.

    Huajian Zhu, Yunrui Cai, Shijia Ma, Yushi Futamura, Jinbiao Li, Wen Zhong, Xiangnan Zhang, Hiroyuki Osada, Hongbin Zou

    ChemSusChem   14 ( 23 ) 5320 - 5327  2021.12  [International journal]

     View Summary

    Bioprivileged molecules hold great promise for supplementing petrochemicals in sustainable organic synthesis of a diverse bioactive products library. Secologanin, a biorenewable monoterpenoid glucoside with unique structural elements, is the key precursor for thousands of natural monoterpenoid alkaloids. Inspired by its inherent highly congested functional groups, a secologanin-based diversity-oriented synthesis (DOS) strategy for novel pseudo-natural alkaloids was developed. All the reactive units of secologanin were involved in these operation simplicity protocols under mild reaction conditions, including the one-step enantioselective transformation of exocyclic C8, C8/C11, and C8/C9/C10 as well as the chemoenzymatic manipulation of endocyclic C2/C6 via the attack by various nucleophiles. A combinatory scenario of the aforementioned reactions further provided diverse polycyclic products with multiple chiral centers. Preliminary activity screening of these newly constructed molecules led to the discovery of antimalarial and highly potent neuroprotective skeletons. The application of green biorenewable secologanin in diversity-oriented pseudo-natural monoterpenoid alkaloid synthesis might encourage the pursuit of valuable bioactive frameworks.

    DOI PubMed

  • 6,9-Dihydroxytetrangulol, a novel angucyclinone antibiotic accumulated in kiqO gene disruptant in the biosynthesis of kinanthraquinone

    Katsuyuki Sakai, Risa Takao, Hiroyuki Koshino, Yushi Futamura, Hiroyuki Osada, Shunji Takahashi

    The Journal of Antibiotics   74 ( 9 ) 593 - 595  2021.09

    DOI PubMed

  • 6,9-Dihydroxytetrangulol, a novel angucyclinone antibiotic accumulated in kiqO gene disruptant in the biosynthesis of kinanthraquinone.

    Katsuyuki Sakai, Risa Takao, Hiroyuki Koshino, Yushi Futamura, Hiroyuki Osada, Shunji Takahashi

    The Journal of antibiotics   74 ( 9 ) 593 - 595  2021.09  [International journal]

     View Summary

    A novel angucyclinone, 6,9-dihydroxytetrangulol, was isolated from Streptomyces lividans TK23 transformed with a kinanthraquinone biosynthetic gene cluster in which the kiqO gene was disrupted. The chemical structure was elucidated by spectroscopic analyses. It showed significant antibacterial activities with an IC50 value of 1.9 μM against Staphylococcus aureus and moderate anticancer activities against HL-60 cells.

    DOI PubMed

  • N-Acetyl-α-hydroxy-β-oxotryptamine, a racemic natural product isolated from Streptomyces sp. 80H647

    Julius Adam V. Lopez, Toshihiko Nogawa, Yushi Futamura, Harumi Aono, Daisuke Hashizume, Hiroyuki Osada

    The Journal of Antibiotics   74 ( 7 ) 477 - 479  2021.07  [Refereed]

    DOI

  • Zealpeptaibolin, an 11-mer cytotoxic peptaibol group with 3 Aib-Pro motifs isolated from Trichoderma sp. RK10-F026

    Mira Syahfriena Amir Rawa, Toshihiko Nogawa, Akiko Okano, Yushi Futamura, Habibah A. Wahab, Hiroyuki Osada

    The Journal of Antibiotics    2021.06  [Refereed]

    DOI

  • Glyoxalase I disruption and external carbonyl stress impair mitochondrial function in human induced pluripotent stem cells and derived neurons

    Tomonori Hara, Manabu Toyoshima, Yasuko Hisano, Shabeesh Balan, Yoshimi Iwayama, Harumi Aono, Yushi Futamura, Hiroyuki Osada, Yuji Owada, Takeo Yoshikawa

    Translational Psychiatry   11 ( 1 ) 275 - 275  2021.06  [Refereed]  [International journal]

     View Summary

    <title>Abstract</title>Carbonyl stress, a specific form of oxidative stress, is reported to be involved in the pathophysiology of schizophrenia; however, little is known regarding the underlying mechanism. Here, we found that disruption of <italic>GLO1</italic>, the gene encoding a major catabolic enzyme scavenging the carbonyl group, increases vulnerability to external carbonyl stress, leading to abnormal phenotypes in human induced pluripotent stem cells (hiPSCs). The viability of <italic>GLO1</italic> knockout (KO)-hiPSCs decreased and activity of caspase-3 was increased upon addition of methylglyoxal (MGO), a reactive carbonyl compound. In the <italic>GLO1</italic> KO-hiPSC-derived neurons, MGO administration impaired neurite extension and cell migration. Further, accumulation of methylglyoxal-derived hydroimidazolone (MG-H1; a derivative of MGO)-modified proteins was detected in isolated mitochondria. Mitochondrial dysfunction, including diminished membrane potential and dampened respiratory function, was observed in the <italic>GLO1</italic> KO-hiPSCs and derived neurons after addition of MGO and hence might be the mechanism underlying the effects of carbonyl stress. The susceptibility to MGO was partially rescued by the administration of pyridoxamine, a carbonyl scavenger. Our observations can be used for designing an intervention strategy for diseases, particularly those induced by enhanced carbonyl stress or oxidative stress.

    DOI PubMed

  • A new peptaibol, RK-026A, from the soil fungus Trichoderma sp. RK10-F026 by culture condition-dependent screening

    Mira Syahfriena Amir Rawa, Toshihiko Nogawa, Akiko Okano, Yushi Futamura, Takemichi Nakamura, Habibah A Wahab, Hiroyuki Osada

    Bioscience, Biotechnology, and Biochemistry   85 ( 1 ) 69 - 76  2021.01  [Refereed]

     View Summary

    <title>Abstract</title>
    A new peptaibol, RK-026A (1) was isolated from a fungus, Trichoderma sp. RK10-F026, along with atroviridin B (2), alamethicin II (3), and polysporin B (4) as a cytotoxic compound, which was selected by principal component analysis of the MS data from 5 different culture conditions. The structure of 1 was determined as a new atroviridin B derivative containing Glu at the 18th residue instead of Gln by NMR and HR-MS analyses including the investigation of detailed MS/MS fragmentations. 1 showed cytotoxicity toward K562 leukemia cells at an IC50 value of 4.1 µm.

    DOI

  • Heterologous Expression of the Biosynthetic Gene Cluster for Verticilactam and Identification of Analogues

    Toshihiko Nogawa, Atsutaka Terai, Keita Amagai, Junko Hashimoto, Yushi Futamura, Akiko Okano, Manabu Fujie, Noriyuki Satoh, Haruo Ikeda, Kazuo Shin-ya, Hiroyuki Osada, Shunji Takahashi

    Journal of Natural Products   83 ( 12 ) 3598 - 3605  2020.12  [Refereed]  [International journal]

     View Summary

    Verticilactam and the new geometric isomers, verticilactams B and C, were produced by heterologous expression of the biosynthetic gene cluster for verticilactam using the Streptomyces avermitilis SUKA17 strain. Only verticilactam, a compound with a characteristic β-ketoamide unit within a 16-membered polyketide macrolactam conjugated with an octalin skeleton, had been previously reported having been isolated from Streptomyces spiroverticillatus JC-8444. In this report, minor verticilactam derivatives were isolated from the transformed strain, and their structures elucidated by spectral analysis. Verticilactam B was a geometric isomer at Δ17 and Δ19, and verticilactam C was the Δ19 and Δ21 isomer. In addition, the absolute configuration of verticilactam was confirmed by ECD analysis and NMR chemical shifts. The stereochemistry assignments of the hydroxy groups at C-10 and C-12 were supported by the domain organization of the polyketide synthase identified in the verticilactam gene cluster. Verticilactam showed moderate activity against the malaria parasite Plasmodium falciparum 3D7 strain with no significant cytotoxicity or antimicrobial effects.

    DOI PubMed

  • Discovery of small-molecule modulator of heterotrimeric Gi-protein by integrated phenotypic profiling and chemical proteomics.

    Tatsuro Kawamura, Yushi Futamura, Erchang Shang, Makoto Muroi, Petra Janning, Masayoshi Ueno, Julian Wilke, Shigeki Takeda, Yasumitsu Kondoh, Slava Ziegler, Nobumoto Watanabe, Herbert Waldmann, Hiroyuki Osada

    Bioscience, biotechnology, and biochemistry     1 - 7  2020.08  [Refereed]  [International journal]

     View Summary

    Discovery of small-molecule inducers of unique phenotypic changes combined with subsequent target identification often provides new insights into cellular functions. Here, we applied integrated profiling based on cellular morphological and proteomic changes to compound screening. We identified an indane derivative, NPD9055, which is mechanistically distinct from reference compounds with known modes of action. Employing a chemical proteomics approach, we then showed that NPD9055 binds subunits of heterotrimeric G-protein Gi. An in vitro [35S]GTPγS-binding assay revealed that NPD9055 inhibited GDP/GTP exchange on a Gαi subunit induced by a G-protein-coupled receptor agonist, but not on another G-protein from the Gαs family. In intact HeLa cells, NPD9055 induced an increase in intracellular Ca2+ levels and ERK/MAPK phosphorylation, both of which are regulated by Gβγ, following its dissociation from Gαi. Our observations suggest that NPD9055 targets Gαi and thus regulates Gβγ-dependent cellular processes, most likely by causing the dissociation of Gβγ from Gαi.

    DOI PubMed

  • Discovery of 3-Cinnamamido-N-Substituted Benzamides as Potential Antimalarial Agents.

    Haicheng Liu, Yushi Futamura, Honghai Wu, Aki Ishiyama, Taotao Zhang, Tao Shi, Qunxiong Zheng, Masato Iwatsuki, Satoshi Ōmura, Hongbin Zou, Hiroyuki Osada

    Medicinal chemistry (Shariqah (United Arab Emirates))    2020.08  [Refereed]  [International journal]

     View Summary

    BACKGROUND: Malaria is one of the most devastating parasitic diseases, yet the discovery of antimalarial agents remains profoundly challenging. Very few new antimalarials have been developed in the past 50 years, while the emergence of drug-resistance continues to appear. OBJECTIVE: This study focuses on the discovery, design, synthesis, and antimalarial evaluation of 3-cinnamamido-N-substituted benzamides. METHOD: In this study, a screening of our compound library was carried out against the multidrug-sensitive Plasmodium falciparum 3D7 strain. Derivatives of the hit were designed, synthesized and tested against P. falciparum 3D7 and the in vivo antimalarial activity of the most active compounds was evaluated using the method of Peters' 4-day suppressive test. RESULTS: The retrieved hit compound 1 containing a 3-cinnamamido-N-substituted benzamide skeleton showed moderate antimalarial activity (IC50 = 1.20 µM) for the first time. A series of derivatives were then synthesized through a simple four-step workflow, and half of them exhibited slightly better antimalarial effect than the precursor 1 during the subsequent in vitro assays. Additionally, compounds 11, 23, 30 and 31 displayed potent activity with IC50 values of approximately 0.1 µM, and weak cytotoxicity against mammalian cells. However, in vivo antimalarial activity is not effective which might be ascribed to the poor solubility of these compounds. CONCLUSION: In this study, phenotypic screen of our compound library resulted in the first report of 3-cinnamamide framework with antimalarial activity and 40 derivatives were then designed and synthesized. Subsequent structure-activity studies showed that compounds 11, 23, 30 and 31 exhibited the most potent and selective activity against P. falciparum 3D7 strain with IC50 values around 0.1 µM. Our work herein sets another example of phenotypic screen-based drug discovery, leading to potentially promising candidates of novel antimalarial agents once given further optimization.

    DOI PubMed

  • Biosynthetic gene cluster identification and biological activity of lucilactaene from Fusarium sp. RK97-94.

    Sho Kato, Takayuki Motoyama, Yushi Futamura, Masakazu Uramoto, Toshihiko Nogawa, Toshiaki Hayashi, Hiroshi Hirota, Akira Tanaka, Naoko Takahashi-Ando, Takashi Kamakura, Hiroyuki Osada

    Bioscience, biotechnology, and biochemistry   84 ( 6 ) 1303 - 1307  2020.06  [Refereed]  [International journal]

     View Summary

    We identified the biosynthetic gene cluster for lucilactaene, a cell cycle inhibitor from a filamentous fungus Fusarium sp. RK 97-94. The luc1 knockout strain accumulated demethylated analogs, indicating the involvement of Luc1 methyltransferase in lucilactaene biosynthesis. Lucilactaene showed potent antimalarial activity. Our data suggested that methylation and ether ring formation are essential for its potent antimalarial activity.

    DOI PubMed

  • Inhibition of mitochondrial complex I by the novel compound FSL0260 enhances high salinity-stress tolerance in Arabidopsis thaliana.

    Kaori Sako, Yushi Futamura, Takeshi Shimizu, Akihiro Matsui, Hiroyuki Hirano, Yasumitsu Kondoh, Makoto Muroi, Harumi Aono, Maho Tanaka, Kaori Honda, Kenshirou Shimizu, Makoto Kawatani, Takeshi Nakano, Hiroyuki Osada, Ko Noguchi, Motoaki Seki

    Scientific reports   10 ( 1 ) 8691 - 8691  2020.05  [Refereed]  [International journal]

     View Summary

    Chemical priming is an attractive and promising approach to improve abiotic stress tolerance in a broad variety of plant species. We screened the RIKEN Natural Products Depository (NPDepo) chemical library and identified a novel compound, FSL0260, enhancing salinity-stress tolerance in Arabidopsis thaliana and rice. Through transcriptome analysis using A. thaliana seedlings, treatment of FSL0260 elevated an alternative respiration pathway in mitochondria that modulates accumulation of reactive oxygen species (ROS). From comparison analysis, we realized that the alternative respiration pathway was induced by treatment of known mitochondrial inhibitors. We confirmed that known inhibitors of mitochondrial complex I, such as rotenone and piericidin A, also enhanced salt-stress tolerance in Arabidopsis. We demonstrated that FSL0260 binds to complex I of the mitochondrial electron transport chain and inhibits its activity, suggesting that inhibition of mitochondrial complex I activates an alternative respiration pathway resulting in reduction of ROS accumulation and enhancement of tolerance to salinity in plants. Furthermore, FSL0260 preferentially inhibited plant mitochondrial complex I rather than a mammalian complex, implying that FSL0260 has a potential to be an agent for improving salt-stress tolerance in agriculture that is low toxicity to humans.

    DOI PubMed

  • Productivity and bioactivity of enokipodins A-D of Flammulina rossica and Flammulina velutipes.

    Akiko Tabuchi, Emi Fukushima-Sakuno, Kumiko Osaki-Oka, Yushi Futamura, Takayuki Motoyama, Hiroyuki Osada, Noemia Kazue Ishikawa, Eiji Nagasawa, Keisuke Tokimoto

    Bioscience, biotechnology, and biochemistry   84 ( 5 ) 876 - 886  2020.05  [Refereed]  [International journal]

     View Summary

    Enokipodins are antimicrobial sesquiterpenes produced by Flammulina velutipes in a mycelial culture medium. To date, enokipodin production has not been reported in other members of the genus Flammulina. Hence, in this study, the production of enokipodins A, B, C, and D by F. velutipes and F. rossica was investigated. Some strains of F. rossica were confirmed to produce at least one of the four enokipodins in the culture medium. However, some strains of F. velutipes did not produce any of the enokipodins. In an antibacterial assay using liquid medium, enokipodin B showed the strongest growth inhibitory activity against Bacillus subtilis among the four types of enokipodins. Enokipodin B inhibited the spore germination of some plant pathogenic fungi. Enokipodins B and D exerted moderate anti-proliferative activity against some cancer cell lines, and enokipodins A and C inhibited the proliferation of the malarial parasite, Plasmodium falciparum.

    DOI PubMed

  • YO-001A, a new antifungal agent produced by Streptomyces sp. YO15-A001.

    Kai Yamamoto, Yushi Futamura, Rachael A Uson-Lopez, Harumi Aono, Takeshi Shimizu, Hiroyuki Osada

    The Journal of antibiotics   72 ( 12 ) 986 - 990  2019.12  [Refereed]  [Domestic journal]

     View Summary

    A new antifungal compound YO-001A was found from the culture broth of Streptomyces sp. YO15-A001, which was isolated from a soil sample collected in Toyama Prefecture. YO-001A was identified through morphological changes-based screening of the rice blast fungus, Pyricularia oryzae (P. oryzae). YO-001A is a new 26-membered macrolide of the oligomycin family, which exhibits potent antifungal activity against P. oryzae with an IC50 of 0.012 µM by disrupting mitochondrial respiration via inhibition of the FOF1-ATPase activity.

    DOI PubMed

  • Structure and biological activity of metarhizin C, a stereoisomer of BR-050 from Tolypocladium album RK17-F0007.

    Nogawa, T, Kawatani, M, Okano, A, Futamura, Y, Aono, H, Shimizu, T, Kato, N, Kikuchi, H, Osada, H

    The Journal of Antibiotics    2019.09  [Refereed]

    DOI

  • Stereocomplementary Chemoenzymatic Pictet-Spengler Reactions for Formation of Rare Azepino-indole Frameworks: Discovery of Antimalarial Compounds

    Cai Yunrui, Shao Nana, Xie Hujun, Futamura Yushi, Panjikar Santosh, Liu Haicheng, Zhu Huajian, Osada Hiroyuki, Zou Hongbin

    ACS CATALYSIS   9 ( 8 ) 7443 - 7448  2019.08  [Refereed]

    DOI

  • Nocardamin glucuronide, a new member of the ferrioxamine siderophores isolated from the ascamycin-producing strain Streptomyces sp. 80H647.

    Lopez JAV, Nogawa T, Futamura Y, Shimizu T, Osada H

    The Journal of antibiotics    2019.08  [Refereed]

    DOI PubMed

  • A novel inhibitor of tumorspheres reveals the activation of the serine biosynthetic pathway upon mitochondrial inhibition.

    Amit Subedi, Makoto Muroi, Yushi Futamura, Tatsuro Kawamura, Harumi Aono, Mayuko Nishi, Akihide Ryo, Nobumoto Watanabe, Hiroyuki Osada

    FEBS letters   593 ( 8 ) 763 - 776  2019.04  [Refereed]  [International journal]

     View Summary

    Differences in the metabolism of cancer cells or cancer stem cells (CSCs) as compared to normal cells have provided avenues to safely target cancers. To discover metabolic inhibitors of CSCs, we performed alkaline phosphatase- and tumoursphere-based drug screening using induced cancer stem cell-like cells. From the screening of a RIKEN NPDepo chemical library, we discovered NPD2381 as a novel and selective cancer-stemness inhibitor that targets mitochondrial metabolism. Using our ChemProteoBase profiling, we found that NPD2381 increases the expression of enzymes within the serine biosynthesis pathway. We also found a role for serine in protecting cancer cells from mitochondrial inhibitors. Our results suggest the existence of a compensatory mechanism to increase the level of intracellular serine in response to mitochondrial inhibitors.

    DOI PubMed

  • Construction of a potato fraction library for the investigation of functional secondary metabolites

    Nogawa Toshihiko, Futamura Yushi, Okano Akiko, Suto Mari, Nakamura Junya, Ishihara Katsuyuki, Osada Hiroyuki

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   83 ( 1 ) 65 - 75  2019  [Refereed]

    DOI

  • Structures and Synthesis of Hitoyopodins: Bioactive Aromatic Sesquiterpenoids Produced by the Mushroom Coprinopsis cinerea (vol 20, pg 6294, 2018)

    Otaka Junnosuke, Shimizu Takeshi, Futamura Yushi, Hashizume Daisuke, Osada Hiroyuki

    ORGANIC LETTERS   20 ( 22 ) 7352  2018.11  [Refereed]

    DOI

  • Catenulisporolides, Glycosylated Triene Macrolides from the Chemically Underexploited Actinomycete Catenulispora Species

    Son S, Hong Y. S, Futamura Y, Jang M, Lee J. K, Heo K. T, Ko S. K, Lee J. S, Takahashi S, Osada H, Jang J. H, Ahn J. S

    Organic Letters   20 ( 22 ) 7234 - 7238  2018.11  [Refereed]

    DOI

  • Structures and Synthesis of Hitoyopodins: Bioactive Aromatic Sesquiterpenoids Produced by the Mushroom Coprinopsis cinerea

    Otaka J, Shimizu T, Futamura Y, Hashizume D, Osada H

    Organic Letters   20 ( 19 ) 6294 - 6297  2018.10  [Refereed]

    DOI

  • Catalytic Enantioselective Synthesis of a Pyrrolizidine-Alkaloid-Inspired Compound Collection with Antiplasmodial Activity

    Jia Z. J, Takayama H, Futamura Y, Aono H, Bauer J. O, Strohmann C, Antonchick A. P, Osada H, Waldmann H

    Journal of Organic Chemistry   83 ( 13 ) 7033 - 7041  2018.07  [Refereed]

    DOI

  • Kinanthraquinone, a new anthraquinone carboxamide isolated from Streptomyces reveromyceticus SN-593-44

    Hiroshi Takagi, Toshihiko Nogawa, Yushi Futamura, Shunji Takahashi, Hiroyuki Osada

    Journal of Antibiotics   71 ( 4 ) 480 - 482  2018.03  [Refereed]

     View Summary

    A new anthraquinone derivative, kinanthraquinone (1) was isolated from Streptomyces reveromyceticus SN-593-44. Its structure was determined by the combination of spectroscopic methods including NMR and MS. Kinanthraquinone had a characteristic carboxamide group and was a rare class of metabolite as an anthraquinone derivative isolated from microbes. It showed moderate cytotoxocity against HL-60 and src ts-NRK cell with IC50 value of 7.9 and 10 μM, respectively.

    DOI

  • Chemical array system, a platform to identify novel hepatitis B virus entry inhibitors targeting sodium taurocholate cotransporting polypeptide.

    Manabu Kaneko, Yushi Futamura, Senko Tsukuda, Yasumitsu Kondoh, Tomomi Sekine, Hiroyuki Hirano, Kento Fukano, Hirofumi Ohashi, Wakana Saso, Ryo Morishita, Satoko Matsunaga, Fumihiro Kawai, Akihide Ryo, Sam-Yong Park, Ryosuke Suzuki, Hideki Aizaki, Naoko Ohtani, Camille Sureau, Takaji Wakita, Hiroyuki Osada, Koichi Watashi

    Scientific reports   8 ( 1 ) 2769 - 2769  2018.02  [Refereed]  [International journal]

     View Summary

    Current anti-hepatitis B virus (HBV) agents including interferons and nucleos(t)ide analogs efficiently suppress HBV infection. However, as it is difficult to eliminate HBV from chronically infected liver, alternative anti-HBV agents targeting a new molecule are urgently needed. In this study, we applied a chemical array to high throughput screening of small molecules that interacted with sodium taurocholate cotransporting polypeptide (NTCP), an entry receptor for HBV. From approximately 30,000 compounds, we identified 74 candidates for NTCP interactants, and five out of these were shown to inhibit HBV infection in cell culture. One of such compound, NPD8716, a coumarin derivative, interacted with NTCP and inhibited HBV infection without causing cytotoxicity. Consistent with its NTCP interaction capacity, this compound was shown to block viral attachment to host hepatocytes. NPD8716 also prevented the infection with hepatitis D virus, but not hepatitis C virus, in agreement with NPD8716 specifically inhibiting NTCP-mediated infection. Analysis of derivative compounds showed that the anti-HBV activity of compounds was apparently correlated with the affinity to NTCP and the capacity to impair NTCP-mediated bile acid uptake. These results are the first to show that the chemical array technology represents a powerful platform to identify novel viral entry inhibitors.

    DOI PubMed

  • Wakodecalines A and B, new decaline metabolites isolated from a fungus Pyrenochaetopsis sp. RK10-F058

    Toshihiko Nogawa, Naoki Kato, Takeshi Shimizu, Akiko Okano, Yushi Futamura, Shunji Takahashi, Hiroyuki Osada

    Journal of Antibiotics   71 ( 1 ) 123 - 128  2018.01  [Refereed]

     View Summary

    Two new decaline metabolites, wakodecalines A and B, were isolated from a fungus, Pyrenochaetopsis sp. RK10-F058, by screening for structurally unique metabolites using LC/MS analysis. Their structures were determined on the basis of NMR and mass spectrometric measurements. The absolute structures were confirmed by a combination of chemical methods including chemical degradation, a modified Mosher's method and Marfey's method, and comparison of the experimental electronic CD (ECD) spectrum with calculated one. Both compounds had a cyclopentanone-fused decaline skeleton and an N-methylated amino acid moiety derived from a serine. They showed moderate antimalarial activity against the Plasmodium falciparum 3D7 strain.

    DOI

  • Discovery of the novel autophagy inhibitor aumitin that targets mitochondrial complex i

    Lucas Robke, Yushi Futamura, Georgios Konstantinidis, Julian Wilke, Harumi Aono, Zhwan Mahmoud, Nobumoto Watanabe, Yao-Wen Wu, Hiroyuki Osada, Luca Laraia, Herbert Waldmann

    Chemical Science   9 ( 11 ) 3014 - 3022  2018  [Refereed]

     View Summary

    Macroautophagy is a conserved eukaryotic process for degradation of cellular components in response to lack of nutrients. It is involved in the development of diseases, notably cancer and neurological disorders including Parkinson's disease. Small molecule autophagy modulators have proven to be valuable tools to dissect and interrogate this crucial metabolic pathway and are in high demand. Phenotypic screening for autophagy inhibitors led to the discovery of the novel autophagy inhibitor aumitin. Target identification and confirmation revealed that aumitin inhibits mitochondrial respiration by targeting complex I. We show that inhibition of autophagy by impairment of mitochondrial respiration is general for several mitochondrial inhibitors that target different mitochondrial complexes. Our findings highlight the importance of mitochondrial respiration for autophagy regulation.

    DOI

  • Bioenergetic and proteomic profiling to screen small molecule inhibitors that target cancer metabolisms

    Yushi Futamura, Makoto Muroi, Harumi Aono, Makoto Kawatani, Marina Hayashida, Tomomi Sekine, Toshihiko Nogawa, Hiroyuki Osada

    Biochimica et Biophysica Acta - Proteins and Proteomics   1867 ( 1 ) 28 - 37  2018  [Refereed]

     View Summary

    Cancer cells can reprogram their metabolic machinery to survive. This altered metabolism, which is distinct from the metabolism of normal cells, is thought to be a possible target for the development of new cancer therapies. In this study, we constructed a screening system that focuses on bioenergetic profiles (specifically oxygen consumption rate and extracellular acidification rate) and characteristic proteomic changes. Thus, small molecules that target cancer-specific metabolism were investigated. We screened the chemical library of RIKEN Natural Products Depository (NPDepo) and found that unantimycin A, which was recently isolated from the fraction library of microbial metabolites, and NPL40330, which is derived from a chemical library, inhibit mitochondrial respiration. Furthermore, we developed an in vitro reconstitution assay method for mitochondrial electron transport chain using semi-intact cells with specific substrates for each complex of the mitochondrial electron transport chain. Our findings revealed that NPL40330 and unantimycin A target mitochondrial complexes I and III, respectively.

    DOI

  • Hitoyol A and B, Two Norsesquiterpenoids from the Basidiomycete Coprinopsis cinerea

    Junnosuke Otaka, Daisuke Hashizume, Yui Masumoto, Atsuya Muranaka, Masanobu Uchiyama, Hiroyuki Koshino, Yushi Futamura, Hiroyuki Osada

    ORGANIC LETTERS   19 ( 15 ) 4030 - 4033  2017.08  [Refereed]

     View Summary

    Hitoyol A (1), an unprecedented norsesquiterpenoid with an exo-tricyclo[S.2.1.0(2,6)]decane skeleton, was isolated from the culture broth of Basidiomycete Coprinopsis cinerea along with a novel skeletal hitoyol B (2) containing 4-cyclopentene-1,3-dione. Their structures and absolute configurations were analyzed by single-crystal X-ray diffraction and electronic circular dichroism spectroscopic methods. Compound 1 is possibly biosynthesized through decarboxylation-induced cyclization of lagopodin B, a known cuparene-type sesquiterpenoid. Compound 2 showed weak antimalarial activity with an IC50 of 59 mu M.

    DOI

  • Trachyspic acid 19-butyl ester, a new inhibitor of Plk1 polo box domain-dependent recognition from uncharacterized fungus RKGS-F2684

    Toshihiko Nogawa, Noriko Ogita, Yushi Futamura, Shigenori Negishi, Nobumoto Watanabe, Hiroyuki Osada

    JOURNAL OF ANTIBIOTICS   70 ( 5 ) 705 - 707  2017.05  [Refereed]

    DOI

  • Mitochondrial uncoupler exerts a synthetic lethal effect against β-catenin mutant tumor cells

    Yuki Shikata, Masaki Kiga, Yushi Futamura, Harumi Aono, Hiroyuki Inoue, Manabu Kawada, Hiroyuki Osada, Masaya Imoto

    Cancer Science   108 ( 4 ) 772 - 784  2017.04  [Refereed]

     View Summary

    The wingless/int-1 (Wnt) signal transduction pathway plays a central role in cell proliferation, survival, differentiation and apoptosis. When β-catenin: a component of the Wnt pathway, is mutated into an active form, cell growth signaling is hyperactive and drives oncogenesis. As β-catenin is mutated in a wide variety of tumors, including up to 10% of all sporadic colon carcinomas and 20% of hepatocellular carcinomas, it has been considered a promising target for therapeutic interventions. Therefore, we screened an in-house natural product library for compounds that exhibited synthetic lethality towards β-catenin mutations and isolated nonactin, an antibiotic mitochondrial uncoupler, as a hit compound. Nonactin, as well as other mitochondrial uncouplers, induced apoptosis selectively in β-catenin mutated tumor cells. Significant tumor regression was observed in the β-catenin mutant HCT 116 xenograft model, but not in the β-catenin wild type A375 xenograft model, in response to daily administration of nonactin in vivo. Furthermore, we found that expression of an active mutant form of β-catenin induced a decrease in the glycolysis rate. Taken together, our results demonstrate that tumor cells with mutated β-catenin depend on mitochondrial oxidative phosphorylation for survival. Therefore, they undergo apoptosis in response to mitochondrial dysfunction following the addition of mitochondrial uncouplers, such as nonactin. These results suggest that targeting mitochondria is a potential chemotherapeutic strategy for tumor cells that harbor β-catenin mutations.

    DOI PubMed

  • Two new triterpenoids from the roots of Pinus densiflora

    Junnosuke Otaka, Masabumi Komatsu, Yasumasa Miyazaki, Yushi Futamura, Hiroyuki Osada

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   81 ( 3 ) 449 - 452  2017.03  [Refereed]

     View Summary

    Chemical investigation of the roots of Pinus densiflora led to the isolation of two new triterpenoids, (24S)-3-methoxy-24,25-epoxy-lanost-9(11)-ene (1) and 29-acetoxy-3-methoxyserrat-14-en-21-ol (2), together with three known serratene-type triterpenoids (3-5) and four known diterpenoids (6-9). Their structures were determined by spectroscopic analyses.

    DOI

  • Opantimycin A, a new metabolite isolated from Streptomyces sp RK88-1355

    Toshihiko Nogawa, Akiko Okano, Chung Liang Lim, Yushi Futamura, Takeshi Shimizu, Shunji Takahashi, Darah Ibrahim, Hiroyuki Osada

    JOURNAL OF ANTIBIOTICS   70 ( 2 ) 222 - 225  2017.02  [Refereed]

    DOI

  • Phenotypic screening meets natural products in drug discovery

    Yushi Futamura, Kai Yamamoto, Hiroyuki Osada

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   81 ( 1 ) 28 - 31  2017.01  [Refereed]

     View Summary

    The Nobel Prize in Physiology or Medicine 2015 was awarded for discoveries related to the control of parasitic diseases using natural products of microbial and plant origin. In current drug discovery programs, synthesized compounds are widely used as a screening source; however, this award reminds us of the importance of natural products. Here, we introduce our phenotypic screening methods based on changes in cell morphology and discuss their effectiveness and impact for natural products in drug discovery.

    DOI

  • RK-144171, a new benadrostin derivative produced by Streptomyces sp RK88-1441

    Jun-Pil Jang, Shunji Takahashi, Yushi Futamura, Toshihiko Nogawa, Jae-Hyuk Jang, Jong Seog Ahn, Hiroyuki Osada

    JOURNAL OF ANTIBIOTICS   70 ( 1 ) 102 - 104  2017.01  [Refereed]

    DOI

  • Octaminomycins A and B, Cyclic Octadepsipeptides Active against Plasmodium falciparum

    Jun-Pil Jang, Toshihiko Nogawa, Yushi Futamura, Takeshi Shimizu, Daisuke Hashizume, Shunji Takahashi, Jae-Hyuk Jang, Jong Seog Ahn, Hiroyuki Osada

    JOURNAL OF NATURAL PRODUCTS   80 ( 1 ) 134 - 140  2017.01  [Refereed]

     View Summary

    Two new cyclic octadepsipeptides, octaminomycins A (1) and B (2), were isolated from a microbial metabolite fraction library of Streptomyces sp. RK85-270 based on Natural Products Plot screening. Their structures were elucidated on the basis of HRESIMS, 1D and 2D NMR spectroscopic data, and MS/MS experiments for sequence analysis. The absolute configurations of the constituent amino acid residues were determined by a combination of single crystal X-ray diffraction and Marfey's methodology. Notably, octaminomycins A (1) and B (2) showed good in vitro antiplasmodial activity against chloroquine-sensitive as well as chloroquine-resistant strains with no cytotoxicity up to 30 mu M.

    DOI

  • Proteomic profiling reveals that collismycin A is an iron chelator

    Makoto Kawatani, Makoto Muroi, Akira Wada, Gyo Inoue, Yushi Futamura, Harumi Aono, Kenshirou Shimizu, Takeshi Shimizu, Yasuhiro Igarashi, Naoko Takahashi-Ando, Hiroyuki Osada

    Scientific Reports   6  2016.12  [Refereed]

     View Summary

    Collismycin A (CMA), a microbial product, has anti-proliferative activity against cancer cells, but the mechanism of its action remains unknown. Here, we report the identification of the molecular target of CMA by ChemProteoBase, a proteome-based approach for drug target identification. ChemProteoBase profiling showed that CMA is closely clustered with di-2-pyridylketone 4,4-dimethyl-3-thiosemicarbazone, an iron chelator. CMA bound to both Fe(II) and Fe(III) ions and formed a 2: 1 chelator-iron complex with a redox-inactive center. CMA-induced cell growth inhibition was completely canceled by Fe(II) and Fe(III) ions, but not by other metal ions such as Zn(II) or Cu(II). Proteomic and transcriptomic analyses showed that CMA affects the glycolytic pathway due to the accumulation of HIF-1a. These results suggest that CMA acts as a specific iron chelator, leading to the inhibition of cancer cell growth.

    DOI

  • Amino-group carrier-protein-mediated secondary metabolite biosynthesis in Streptomyces

    Fumihito Hasebe, Kenichi Matsuda, Taro Shiraishi, Yushi Futamura, Takeshi Nakano, Takeo Tomita, Ken Ishigami, Hikari Taka, Reiko Mineki, Tsutomu Fujimura, Hiroyuki Osada, Tomohisa Kuzuyama, Makoto Nishiyama

    NATURE CHEMICAL BIOLOGY   12 ( 11 ) 967 - +  2016.11  [Refereed]

     View Summary

    Amino-group carrier proteins (AmCPs) mediate the biosynthesis of lysine and arginine in some bacteria and archaea. Here we demonstrate that an uncharacterized AmCP-mediated biosynthetic system functions to biosynthesize the previously uncharacterized and nonproteinogenic amino acid (2S,6R)-diamino-(5R,7)-dihydroxy-heptanoic acid (DADH) in Streptomyces sp. SANK 60404. DADH is incorporated into a novel peptide metabolite, vazabitide A, featuring an azabicyclo-ring structure, by nonribosomal peptide synthetases and successive modification enzymes in this bacterium. As the AmCP-mediated machinery for DADH biosynthesis is widely distributed in bacteria, further analysis of uncharacterized AmCP-containing gene clusters will lead to the discovery of novel bioactive compounds and novel biosynthetic enzymes.

    DOI

  • High-throughput screening identifies artesunate as selective inhibitor of cancer sternness: Involvement of mitochondrial metabolism

    Amit Subedi, Yushi Futamura, Mayuko Nishi, Akihide Ryo, Nobumoto Watanabe, Hiroyuki Osada

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   477 ( 4 ) 737 - 742  2016.09  [Refereed]

     View Summary

    Cancer stem cells (CSCs) have robust systems to maintain cancer stemness and drug resistance. Thus, targeting such robust systems instead of focusing on individual signaling pathways should be the approach allowing the identification of selective CSC inhibitors. Here, we used the alkaline phosphatase (ALP) assay to identify inhibitors for cancer sternness in induced cancer stem-like (iCSCL) cells. We screened several compounds from natural product chemical library and evaluated hit compounds for their efficacy on cancer stemness in iCSCL tumorspheres. We identified artesunate, an antimalarial drug, as a selective inhibitor of cancer stemness. Artesunate induced mitochondrial dysfunction that selectively inhibited cancer stemness of iCSCL cells, indicating an essential role of mitochondrial metabolism in cancer sternness. (C) 2016 Elsevier Inc. All rights reserved.

    DOI

  • Unantimycin A, a new neoantimycin analog isolated from a microbial metabolite fraction library

    Chung Liang Lim, Toshihiko Nogawa, Akiko Okano, Yushi Futamura, Makoto Kawatani, Shunji Takahashi, Darah Ibrahim, Hiroyuki Osada

    JOURNAL OF ANTIBIOTICS   69 ( 6 ) 456 - 458  2016.06  [Refereed]

    DOI

  • Proteomic profiling of small-molecule inhibitors reveals dispensability of MTH1 for cancer cell survival

    Tatsuro Kawamura, Makoto Kawatani, Makoto Muroi, Yasumitsu Kondoh, Yushi Futamura, Harumi Aono, Miho Tanaka, Kaori Honda, Hiroyuki Osada

    SCIENTIFIC REPORTS   6  2016.05  [Refereed]

     View Summary

    Since recent publications suggested that the survival of cancer cells depends on MTH1 to avoid incorporation of oxidized nucleotides into the cellular DNA, MTH1 has attracted attention as a potential cancer therapeutic target. In this study, we identified new purine-based MTH1 inhibitors by chemical array screening. However, although the MTH1 inhibitors identified in this study targeted cellular MTH1, they exhibited only weak cytotoxicity against cancer cells compared to recently reported first-in-class inhibitors. We performed proteomic profiling to investigate the modes of action by which chemically distinct MTH1 inhibitors induce cancer cell death, and found mechanistic differences among the first-in-class MTH1 inhibitors. In particular, we identified tubulin as the primary target of TH287 and TH588 responsible for the antitumor effects despite the nanomolar MTH1-inhibitory activity in vitro. Furthermore, overexpression of MTH1 did not rescue cells from MTH1 inhibitor-induced cell death, and siRNA-mediated knockdown of MTH1 did not suppress cancer cell growth. Taken together, we conclude that the cytotoxicity of MTH1 inhibitors is attributable to off-target effects and that MTH1 is not essential for cancer cell survival.

    DOI

  • New Cyclic Lipopeptides of the Iturin Class Produced by Saltern-Derived Bacillus sp KCB14S006

    Sangkeun Son, Sung-Kyun Ko, Mina Jang, Jong Won Kim, Gil Soo Kim, Jae Kyoung Lee, Eun Soo Jeon, Yushi Futamura, In-Ja Ryoo, Jung-Sook Lee, Hyuncheol Oh, Young-Soo Hong, Bo Yeon Kim, Shunji Takahashi, Hiroyuki Osada, Jae-Hyuk Jang, Jong Seog Ahn

    MARINE DRUGS   14 ( 4 )  2016.04  [Refereed]

     View Summary

    Salterns, one of the most extreme natural hypersaline environments, are a rich source of halophilic and halotolerant microorganisms, but they remain largely underexplored ecological niches in the discovery of bioactive secondary metabolites. In continued efforts to investigate the metabolic potential of microbial populations from chemically underexplored sites, three new lipopeptides named iturin F-1, iturin F-2 and iturin A(9) (1-3), along with iturin A(8) (4), were isolated from Bacillus sp. KCB14S006 derived from a saltern. The structures of the isolated compounds were established by 1D-, 2D-NMR and HR-ESIMS, and their absolute configurations were determined by applying advanced Marfey's method and CD spectroscopy. All isolates exhibited significant antifungal activities against various pathogenic fungi and moderate cytotoxic activities toward HeLa and src(ts)-NRK cell lines. Moreover, in an in vitro enzymatic assay, compound 4 showed a significant inhibitory activity against indoleamine 2,3-dioxygenase.

    DOI

  • Structures and biological activities of azaphilones produced by Penicillium sp KCB11A109 from a ginseng field

    Sangkeun Son, Sung-Kyun Ko, Jong Won Kim, Jae Kyoung Lee, Mina Jang, In-Ja Ryoo, Gwi Ja Hwang, Min Cheol Kwon, Kee-Sun Shin, Yushi Futamura, Young-Soo Hong, Hyuncheol Oh, Bo Yeon Kim, Masashi Ueki, Shunji Takahashi, Hiroyuki Osada, Jae-Hyuk Jang, Jong Seog Ahn

    PHYTOCHEMISTRY   122   154 - 164  2016.02  [Refereed]

     View Summary

    Twelve metabolites, including five highly oxygenated azaphilones, geumsanols A-E, along with seven known analogues were isolated from Penicillium sp. KCB11A109, a fungus derived from a ginseng field. Their structures were assigned by spectroscopic means (NMR and MS), and stereochemistries were determined by extensive spectroscopic analyses (H-1 -H-1 coupling constants, NOESY, and HETLOC) and chemical derivatizations (modified Mosher's method and acetonide formation). The isolates were evaluated for their anticancer, antimicrobial, antimalarial activities, and phenotypic effects in zebrafish development. Of these compounds possessing no pyranoquinone core, only geumsanol E exhibited cytotoxic activities and toxic effects on zebrafish embryos, suggesting that a double bond at C-11 and C-12 is important for biological activity. (C) 2015 Elsevier Ltd. All rights reserved.

    DOI

  • An overproduction of astellolides induced by genetic disruption of chromatin-remodeling factors in Aspergillus oryzae

    Yasutomo Shinohara, Makoto Kawatani, Yushi Futamura, Hiroyuki Osada, Yasuji Koyama

    JOURNAL OF ANTIBIOTICS   69 ( 1 ) 4 - 8  2016.01  [Refereed]

     View Summary

    The filamentous fungus Aspergillus oryzae is an important industrial mold. Recent genomic analysis indicated that A. oryzae has a large number of biosynthetic genes for secondary metabolites (SMs), but many of the SMs they produce have not been identified. For better understanding of SMs production by A. oryzae, we screened a gene-disruption library of transcription factors including chromatin-remodeling factors and found two gene disruptions that show similarly altered SM production profiles. One is a homolog of Aspergillus nidulans cclA, a component of the histone 3 lysine 4 (H3K4) methyltransferase complex of proteins associated with Set1 complex, and the other, sppA, is an ortholog of Saccharomyces cerevisiae SPP1, another component of a complex of proteins associated with Set1 complex. The cclA and sppA disruptions in A. oryzae are deficient in trimethylation of H3K4. Furthermore, one of the SMs that increased in the cclA disruptant was identified as astellolide F (14-deacetyl astellolide B). These data indicate that both cclA and sppA affect production of SMs including astellolides by affecting the methylation status of H3K4 in A. oryzae.

    DOI

  • Integrated profiling methods for identifying the targets of bioactive compounds: MorphoBase and ChemProteoBase

    Makoto Muroi, Yushi Futamura, Hiroyuki Osada

    NATURAL PRODUCT REPORTS   33 ( 5 ) 621 - 625  2016  [Refereed]

     View Summary

    Many useful compounds from natural products have been discovered through phenotype-based screening. However, the target identification process for compounds is laborious and time-consuming. With the development of new equipment and methodologies for biological analyses, a variety of profiling methods that utilize large sets of experimental data have been established. Here, we highlight the utility of our identification approaches, MorphoBase and ChemProteoBase.

    DOI

  • Methyl 3-((6-Methoxy-1,4-dihydroindeno[1,2-c]pyrazol-3-yl)amino)benzoate (GN39482) as a Tubulin Polymerization Inhibitor Identified by MorphoBase and ChemProteoBase Profiling Methods

    Hidemitsu Minegishi, Yushi Futamura, Shinji Fukashiro, Makoto Muroi, Makoto Kawatani, Hiroyuki Osada, Hiroyuki Nakamura

    JOURNAL OF MEDICINAL CHEMISTRY   58 ( 10 ) 4230 - 4241  2015.05  [Refereed]

     View Summary

    A series of indenopyrazoles was synthesized from the corresponding indanones arid phenyl isothiocyanates in two steps. Among the compounds synthesized, methyl 3-((6-methoxy-1,4-dihydroindeno[1,2-c]pyrazol-3-yl)amino)benzoate 6m (GN39482) was found to possess a promising antiproliferative activity toward human cancer cells without affecting any antimicrobial and antimalarial activities at 100 nM. Both a methoxy group at R-1 position and a methoxycarbonyl group at R-2 position of the anilinoquinazoline framework are essential for the high cell growth inhibition. Both MorphoBase and ChemProteoBase profiling analyses suggested that compound 6m was classified as a tubulin inhibitor. Indeed, compound 6m inhibited the acetylated tubulin accumulation and the microtubule formation and induced G2/M cell. Cycle arrest in HeLa cells, revealing that a promising aritiproliferative activity of compound 6m toward human cancer cells is probably caused by the tubulin polymerization inhibition.

    DOI

  • RK-270A-C, new oxindole derivatives isolated from a microbial metabolites fraction library of Streptomyces sp RK85-270

    Jun-Pil Jang, Toshihiko Nogawa, Masakazu Uramoto, Akiko Okano, Yushi Futamura, Takeshi Shimizu, Shunji Takahashi, Jae-Hyuk Jang, Jong Seog Ahn, Hiroyuki Osada

    JOURNAL OF ANTIBIOTICS   68 ( 4 ) 293 - 295  2015.04  [Refereed]

    DOI

  • Synthesis and biological activity of 5-(4-methoxyphenyl)-oxazole derivatives

    Daisuke Yamamuro, Ryuji Uchida, Masaki Ohtawa, Shiho Arima, Yushi Futamura, Masumi Katane, Hiroshi Homma, Tohru Nagamitsu, Hiroyuki Osada, Hiroshi Tomoda

    BIOORGANIC & MEDICINAL CHEMISTRY LETTERS   25 ( 2 ) 313 - 316  2015.01  [Refereed]

     View Summary

    5-(4'-Methoxyphenyl)-oxazole (MPO), originally reported as a synthetic compound, was isolated from fungal culture broth as an inhibitor of hatch and growth of Caenorhabditis elegans. Nineteen MPO derivatives were chemically synthesized, but showed no effect on C. elegans hatch and growth. These findings strongly suggested that the whole structure of MPO is essential for anti-C. elegans activity. (C) 2014 Elsevier Ltd. All rights reserved.

    DOI

  • Alkyldihydropyrones, new polyketides synthesized by a type III polyketide synthase from Streptomyces reveromyceticus

    Teruki Aizawa, Seung-Young Kim, Shunji Takahashi, Masahiko Koshita, Mioka Tani, Yushi Futamura, Hiroyuki Osada, Nobutaka Funa

    JOURNAL OF ANTIBIOTICS   67 ( 12 ) 819 - 823  2014.12  [Refereed]

     View Summary

    Genome sequencing allows a rapid and efficient identification of novel catalysts that produce novel secondary metabolites. Here we describe the catalytic properties of dihydropyrone synthase A (DpyA), a novel type III polyketide synthase encoded in a linear plasmid of Streptomyces reveromyceticus. Heterologous expression of dpyA led to the accumulation of alkyldihydropyrones A (1), B (2), C (3) and D (4), which are novel dihydropyran compounds that exhibit weak cytotoxicity against the leukemia cell line HL-60. DpyA catalyzes the condensation of beta-hydroxyl acid thioester and methylmalonyl-CoA to yield a triketide intermediate that then undergoes lactonization of a secondary alcohol and a thioester to give alkyldihydropyrone.

    DOI

  • Identification of a molecular target of a novel fungal metabolite, pyrrolizilactone, by phenotypic profiling systems

    Yushi Futamura, Makoto Kawatani, Makoto Muroi, Harumi Aono, Toshihiko Nogawa, Hiroyuki Osada

    ChemBioChem   14 ( 18 ) 2456 - 2463  2013.12  [Refereed]

     View Summary

    In the course of screening our microbial metabolite fraction library, we identified a novel pyrrolizidinone compound, pyrrolizilactone. In this study, we report the identification and characterization of a molecular target for pyrrolizilactone by using two phenotypic profiling systems. Cell morphology-based profiling analysis using an imaging cytometer (MorphoBase) classified pyrrolizilactone as a proteasome inhibitor. Consistently, proteome-based profiling analysis using 2D difference gel electrophoresis (DIGE
    ChemProteoBase) also demonstrated that pyrrolizilactone is associated with proteasome inhibition. On the basis of these predictions, we determined that pyrrolizilactone is a novel type of proteasome inhibitor inhibiting the trypsin-like activity of the proteasome. A novel trypsin-like proteasome inhibitor: Pyrrolizilactone, a novel fungal metabolite, was identified from our microbial metabolite fraction library. With the aid of two phenotype profiling systems - MorphoBase and ChemProteoBase - we established that pyrrolizilactone is a unique proteasome inhibitor targeting trypsin-like activity of the proteasome. Copyright © 2013 WILEY-VCH Verlag GmbH &amp
    Co. KGaA, Weinheim.

    DOI PubMed

  • Identification of small molecule inhibitors of p27(Kip1) ubiquitination by high-throughput screening

    Li-Ching Ooi, Nobumoto Watanabe, Yushi Futamura, Shaida Fariza Sulaiman, Ibrahim Darah, Hiroyuki Osada

    CANCER SCIENCE   104 ( 11 ) 1461 - 1467  2013.11  [Refereed]

     View Summary

    Dysregulation of p27(Kip1) due to proteolysis that involves the ubiquitin ligase (SCF) complex with S-phase kinase-associated protein 2 (Skp2) as the substrate-recognition component (SCFSkp2) frequently results in tumorigenesis. In this report, we developed a high-throughput screening system to identify small-molecule inhibitors of p27(Kip1) degradation. This system was established by tagging Skp2 with fluorescent monomeric Azami Green (mAG) and CDK subunit 1 (Cks1) (mAGSkp2-Cks1) to bind to p27(Kip1) phosphopeptides. We identified two compounds that inhibited the interaction between mAGSkp2-Cks1 and p27(Kip1): linichlorin A and gentian violet. Further studies have shown that the compounds inhibit the ubiquitination of p27(Kip1)in vitro as well as p27(Kip1) degradation in HeLa cells. Notably, both compounds exhibited preferential antiproliferative activity against HeLa and tsFT210 cells compared with NIH3T3 cells and delayed the G(1) phase progression in tsFT210 cells. Our approach indicates a potential strategy for restoring p27(Kip1) levels in human cancers.

    DOI

  • Pyrrolizilactone, a new pyrrolizidinone metabolite produced by a fungus

    Toshihiko Nogawa, Makoto Kawatani, Masakazu Uramoto, Akiko Okano, Harumi Aono, Yushi Futamura, Hiroyuki Koshino, Shunji Takahashi, Hiroyuki Osada

    JOURNAL OF ANTIBIOTICS   66 ( 10 ) 621 - 623  2013.10  [Refereed]

    DOI

  • Morphobase, an Encyclopedic Cell Morphology Database, and Its Use for Drug Target Identification

    Yushi Futamura, Makoto Kawatani, Sayaka Kazami, Kenichi Tanaka, Makoto Muroi, Takeshi Shimizu, Koji Tomita, Nobumoto Watanabe, Hiroyuki Osada

    CHEMISTRY & BIOLOGY   19 ( 12 ) 1620 - 1630  2012.12  [Refereed]

     View Summary

    Visual observation is a powerful approach for screening bioactive compounds that can facilitate the discovery of attractive druggable targets following their chemicobiological validation. So far, many high-content approaches, using sophisticated imaging technology and bioinformatics, have been developed. In our study, we aimed to develop a simpler method that focuses on intact cell images because we found that dynamic changes in morphology are informative, often reflecting the mechanism of action of a drug. Here, we constructed a chemical-genetic phenotype profiling system, based on the high-content cell morphology database Morphobase. This database compiles the phenotypes of cancer cell lines that are induced by hundreds of reference compounds, wherein those of well-characterized anticancer drugs are classified by mode of action. Furthermore, we demonstrate the applicability of this system in identifying NPD6689, NPD8617, and NPD8969 as tubulin inhibitors.

    DOI

  • Comprehensive predictions of target proteins based on protein-chemical interaction using virtual screening and experimental verifications

    Hiroki Kobayashi, Hiroko Harada, Masaomi Nakamura, Yushi Futamura, Akihiro Ito, Minoru Yoshida, Shun-Ichiro Iemura, Kazuo Shin-Ya, Takayuki Doi, Takashi Takahashi, Tohru Natsume, Masaya Imoto, Yasubumi Sakakibara

    BMC Chemical Biology   12   2  2012  [Refereed]

     View Summary

    Background: Identification of the target proteins of bioactive compounds is critical for elucidating the mode of action
    however, target identification has been difficult in general, mostly due to the low sensitivity of detection using affinity chromatography followed by CBB staining and MS/MS analysis. Results: We applied our protocol of predicting target proteins combining in silico screening and experimental verification for incednine, which inhibits the anti-apoptotic function of Bcl-xL by an unknown mechanism. One hundred eighty-two target protein candidates were computationally predicted to bind to incednine by the statistical prediction method, and the predictions were verified by in vitro binding of incednine to seven proteins, whose expression can be confirmed in our cell system.As a result, 40% accuracy of the computational predictions was achieved successfully, and we newly found 3 incednine-binding proteins. Conclusions: This study revealed that our proposed protocol of predicting target protein combining in silico screening and experimental verification is useful, and provides new insight into a strategy for identifying target proteins of small molecules. © 2012 Kobayashi et al.
    licensee BioMed Central Ltd.

    DOI PubMed

  • Vipirinin, a Coumarin-based HIV-1 Vpr Inhibitor, Interacts with a Hydrophobic Region of VPR

    Eugene Boon Beng Ong, Nobumoto Watanabe, Akiko Saito, Yushi Futamura, Khaled Hussein Abd El Galil, Atsushi Koito, Nazalan Najimudin, Hiroyuki Osada

    JOURNAL OF BIOLOGICAL CHEMISTRY   286 ( 16 ) 14049 - 14056  2011.04  [Refereed]

     View Summary

    The human immunodeficiency virus 1 (HIV-1) viral protein R (Vpr) is an accessory protein that has been shown to have multiple roles in HIV-1 pathogenesis. By screening chemical libraries in the RIKEN Natural Products Depository, we identified a 3-phenyl coumarin-based compound that inhibited the cell cycle arrest activity of Vpr in yeast and Vpr-dependent viral infection of human macrophages. We determined its minimal pharmacophore through a structure-activity relationship study and produced more potent derivatives. We detected direct binding, and by assaying a panel of Vpr mutants, we found the hydrophobic region about residues Glu-25 and Gln-65 to be potentially involved in the binding of the inhibitor. Our findings exposed a targeting site on Vpr and delineated a convenient approach to explore other targeting sites on the protein using small molecule inhibitors as bioprobes.

    DOI

  • Vacuolar H+-ATPase inhibitors overcome Bcl-xL-mediated chemoresistance through restoration of a caspase-independent apoptotic pathway

    Sasazawa Yukiko, Futamura Yushi, Tashiro Etsu, Imoto Masaya

    CANCER SCIENCE   100 ( 8 ) 1460 - 1467  2009.08  [Refereed]

    DOI

  • 1 Isolation, Structure Elucidation, Biological Activities, and SAR Study of Trierixin and Quinotrierixin, Novel Small Molecule Inhibitors of ER Stress-induced XBP1 Activation(Oral Presentation)

    Kawamura Tatsuro, Tashiro Etsu, Futamura Yushi, Shindo Kazutoshi, Imoto Masaya

    Symposium on the Chemistry of Natural Products, symposium papers   ( 50 ) 29 - 34  2008.09

     View Summary

    In cytotoxic conditions such as hypoxia, nutrient deprivation and low pH, unfolded proteins accumulate in cells, leading to ER stress. Poorly vascularized solid-tumor cells are considered to adapt to ER stress by activating a transcription factor XBP1, resulting in tumor survival. Therefore, inhibitors of XBP1 activation would be a new type of anticancer drugs. To screen for inhibitors of ER stress-induced XBP1 activation, we established a screening system (Fig. 1), and isolated novel triene-ansamycin group compounds trierixin and quinotrierixin from culture broths of Streptomyces sp. AC654 and Streptomyces sp. PAE37, respectively (Fig. 3). The planar structures of trierixin and quinotrierixin were elucidated on the basis of the spectroscopical properties (Fig. 2). Trierixin inhibited ER stress-induced XBP1 activation and cell growth of HeLa cells with IC_<50> of 28nM and 10nM, respectively. Quinotrierixin also showed both inhibitory activities with the same dose range. Therefore, we proposed a hypothesis that triene-ansamycin group compounds inhibited tumor cell growth via inhibition of XBP1 activation. To confirm this possibility, we isolated six additional triene-ansamycin group compounds (including three novel compounds) from the culture broth of Streptomyces sp. PAE37 by referring to the characteristic UV spectra (λ_<max> 260, 270 and 280nm), and prepared four derivatives by chemical reactions on the natural products (Fig. 3, Fig. 4). Using these compounds, we performed SAR study of triene-ansamycin group compounds. As a result, there was high correlation between inhibitory effects of triene-ansamycin group compounds against XBP1 activation and those against tumor cell growth (Fig. 5).

    DOI CiNii

  • The identification of an osteoclastogenesis inhibitor through the inhibition of glyoxalase I

    Makoto Kawatani, Hideo Okumura, Kaori Honda, Naoki Kanoh, Makoto Muroi, Naoshi Dohmae, Masamichi Takami, Mitsuhiro Kitagawa, Yushi Futamura, Masaya Imoto, Hiroyuki Osada

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   105 ( 33 ) 11691 - 11696  2008.08  [Refereed]

     View Summary

    Osteoclasts, bone-resorptive multinucleated cells derived from hematopoietic stem cells, are associated with many bone-related diseases, such as osteoporosis. Osteoclast-targeting small-molecule inhibitors are valuable tools for studying osteoclast biology and for developing antiresorptive agents. Here, we have discovered that methyl-gerfelin (M-GFN), the methyl ester of the natural product gerfelin, suppresses osteoclastogenesis. By using M-GFN-immobilized beads, glyoxalase 1 (GLO1) was identified as an M-GFN-binding protein. GLO1 knockdown and treatment with an established GLO1 inhibitor in osteoclast progenitor cells interfered with osteoclast generation, suggesting that GLO1 activity is required for osteoclastogenesis. In cells, GLO1 plays a critical role in the detoxification of 2-oxoaldehydes, such as methylglyoxal. M-GFN inhibited the enzymatic activity of GLO1 in vitro and in situ. Furthermore, the cocrystal structure of the GLO1/M-GFN complex revealed the binding mode of M-GFN at the active site of GLO1. These results suggest that M-GFN targets GLO1, resulting in the inhibition of osteoclastogenesis.

    DOI

  • Discovery of incednine as a potent modulator of the anti-apoptotic function of Bcl-xL from microbial origin

    Yushi Futamura, Ryuichi Sawa, Yoji Umezawa, Masayuki Igarashi, Hikaru Nakamura, Kimiko Hasegawa, Mikio Yarnasaki, Etsu Tashiro, Yoshikazu Takahashi, Yuzuru Akarnatsu, Masaya Imoto

    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY   130 ( 6 ) 1822 - +  2008.02  [Refereed]

     View Summary

    Anti-apoptotic oncoproteins Bcl-2 and BcI-xL are overexpressed in many cancers and play a crucial role in cancer initiation, progression, and resistance to chemotherapy. Therefore, the discovery of a functional inhibitor for these proteins and improved understanding of the molecular mechanisms of these proteins will be an aid to novel anti-tumor therapies. Here, using chemical-genetic cell-based screening, we have discovered a chemically and biologically unique substance, incednine, as a novel functional modulator of Bcl-2/Bcl-xL from the fermentation broth of Streptomyces sp. ML-693-90F3. This compound was isolated as a HCI salt by solvent extraction and using centrifugal liquid-liquid partition chromatography. Its structure was elucidated by spectroscopic analysis, X-ray crystallographic analysis, and computational studies. Incednine has a molecular formula, C(42)H(63)N(3)O(8), and consists of a novel skeletal structure, enol-ether amide in a 24-membered macrolactam core, with two aminosugars. Bcl-xL-overexpressing Ms-1 cells displayed resistance to several anti-tumor agents; however, anti-tumor agent-induced cell death was observed only when cells were treated with incednine. Overexpression of BcI-xL inhibited cell death in Bax-overexpressing HEK293T cells by forming a complex with Bax, whereas Bcl-xL failed to inhibit cell death in the presence of incednine without affecting the heterodimerization of Bcl-xL and Bax. These findings suggest that incednine serves as a potent modulator of the anti-apoptotic Bcl-2/Bcl-xL distinct from the other known Bcl-2 inhibitors and could provide a chemical probe to study the underlying mechanisms of Bcl-2/Bcl-xL.

    DOI

  • Trierixin, a novel inhibitor of ER stress-induced XBP1 activation from Streptomyces sp - II. structure elucidation

    Yushi Futamura, Etsu Tashiro, Naoka Hironiwa, Jun Kohno, Maki Nishio, Kazutoshi Shindo, Masaya Imoto

    JOURNAL OF ANTIBIOTICS   60 ( 9 ) 582 - 585  2007.09  [Refereed]

     View Summary

    Trierixin, a new member of the triene-ansamycin group, has been isolated from the fermentation broth of Streptomyces sp. AC654 as an inhibitor of ER stress-induced XBP1 activation. The structure of trierixin was determined on the basis of its spectroscopical and chemical properties. Trierixin possessed a 21-membered macrocyclic lactam, which contains a methylthio-benzenediol structure, and a cyclohexanecarbonylalanine moiety. Trierixin is thus elucidated as 21-thiomethylmycotrienin II.

    DOI

  • Trierixin, a novel inhibitor of ER stress-induced XBP1 activation from Streptomyces sp. I. Taxonomy, fermentation, isolation, and biological activities

    Tashiro E, Hironiwa N, Kitagawa M, Futamura Y, Suzuki S, Nishio M, Imoto M

    Journal of Antibiotics   60 ( 9 ) 547 - 553  2007.09  [Refereed]

     View Summary

    In the course of screening for an inhibitor of ER stress-induced XBP1 activation, we isolated a new member of the triene-ansamycin group compound, trierixin, from a culture broth of Streptomyces sp. AC 654. Trierixin was purified by column chromatography on silica gel and by HPLC. The molecular formula of trierixin is C37H52N2O8S. Trierixin inhibited thapsigargin-induced XBP1-luciferase activation in HeLa/XBP1-luc cells and endogenous XBP1 splicing in HeLa cells with an IC 50 of 14 ng/ml and 19 ng/ml, respectively. Moreover, in the process of isolating trierixin, we isolated structurally related mycotrienin II and trienomycin A as inhibitors of ER stress-induced XBP1 activation from a culture broth of a trierixin-producing strain. This study provides the first observation that triene-ansamycins have a novel inhibitory effect against XBP1 activation. © Japan Antibiotics Research Association.

    DOI PubMed

  • 38 Studies on Incednine, a Novel Inactivator of Anti-apoptotic Bcl-2 : Isolation, Structure Elucidation, Biological Activities

    Futamura Yushi, Sawa Ryuichi, Umezawa Yoji, Tashiro Etsu, Igarashi Masayuki, Takahashi Yoshikazu, Akamatsu Yuzuru, Imoto Masaya

    Symposium on the Chemistry of Natural Products, symposium papers   ( 47 ) 193 - 198  2005.09

     View Summary

    It is well known that the anti-apoptotic protein Bcl-2 and its homologue Bcl-X_L are overexpressed in many human cancers and play a crucial role in cancer progression. Therefore, the functional blockade of Bcl-2/Bcl-X_L will be a promise for novel anticancer therapeutics by way of improving tumorigenesis. In the course of screening for an inhibitor of Bcl-X_L, anti-apoptotic function using human SCLC Ms-1 cells that overexpress Bcl-X_L, we found Incednine (1) in the fermentation broth of the strain Streptomyces sp. ML694-90F3. 1 was isolated as a HCl salt by using the Centrifugal liquid-liquid Partition Chromatography, effectively. The planar structure of 1, C_<42>H_<63>N_3O_8 ([M+H]^+, m/z 738.4680, Δ-1.3mmu), was elucidated on the basis of the various NMR spectra data including TOCSY experiments (Fig. 1). The relative stereochemistry of 1 was determined by the analysis of NOE experiments and coupling constants (Fig. 2). The conformation of the aglycon was confirmed by Discovery III-computations. The absolute stereochemistry of 1 was established by modified Mosher's method and X-ray crystallographic analysis (Fig. 3, 4). Incednine has a unique skeletal structure, "enol-ether amide" in the 24-membered macrolactum core, with an attachment of a disaccharide. Bcl-X_L-overexpressing Ms-1 cells displayed resistance to several anti-tumor agents, however, anti-tumor agents-induced cell death was observed only when cells were treated with 1. Overexpression of Bcl-X_L inhibited cell death in Bax-overexpressing HEK293T cells by forming a complex with Bcl-X_L and Bax, whereas it failed to inhibit cell death in the presence of 1 without affecting the formation of Bcl-X_L and Bax complex. The inhibitory mechanism of anti-apoptotic Bcl-X_L by 1 is now under investigation.

    DOI CiNii

▼display all

Misc

  • 抗生物質研究からケミカルバイオロジー研究への変遷

    長田 裕之, 野川 俊彦, 二村 友史

    MEDCHEM NEWS   25   16 - 21  2015.02  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

  • A Target Identification System Based on MorphoBase, ChemProteoBase, and Photo-Cross-Linking Beads.

    Osada Hiroyuki, Makoto Muroi, Yasumitsu Kondoh, Yushi Futamura

    Concepts and Case Studies in Chemical Biology (Ed. Waldmann H, Janning P)     163 - 176  2014.06  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

  • 新たな抗がん剤探索を加速する新スクリーニング技術

    長田 裕之, 二村 友史, 渡辺 信元, 近藤 恭光

    細胞工学   32   655 - 659  2013.06  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

  • 創薬基盤となる化合物リソースの整備と活用

    二村 友史, 河村 達郎, 長田 裕之

    腫瘍内科     321 - 331  2012.03  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

  • Vacuolar H^+-ATPase inhibitors overcome Bcl-xL-mediated chemoresistance through restoration of a caspase-independent apoptotic pathway

    SASAZAWA Yukiko, FUTAMURA Yushi, TASHIRO Etsu, IMOTO Masaya

      100 ( 8 ) 1460 - 1467  2009.08

    CiNii

  • 分子標的治療薬各論 細胞周期阻害剤

    二村 友史, 井本 正哉

    がん分子標的治療 (鶴尾隆 編)     308 - 313  2008.09  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

  • 分子標的治療のターゲット 1. 細胞増殖

    二村 友史, 井本 正哉

    臨床婦人科産科   61   1224 - 1229  2007.10  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

  • 微生物代謝産物を用いたケミカルバイオロジー

    二村 友史, 井本 正哉

    バイオインダストリー   24   30 - 38  2007.07  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

▼display all

Awards

  • 平成26年度 日本がん分子標的治療学会 研究奨励賞

    2015.06   日本がん分子標的治療学会  

    Winner: 二村 友史

  • 平成25年度 理研研究奨励賞

    2014.03   理化学研究所  

    Winner: 二村 友史

  • GE Life Sciences Day 2013 Poster Award

    2013.07   GE Healthcare Japan Corporation  

    Winner: 二村 友史

  • 平成22年度 日本がん分子標的治療学会 特別賞

    2010.07   日本がん分子標的治療学会  

    Winner: 二村 友史

Research Projects

  • 人工知能を活用した抗真菌剤の開発研究

    日本学術振興会  科学研究費助成事業 基盤研究(A)

    Project Year :

    2021.04
    -
    2024.03
     

    二村 友史

  • カンジダ二形性制御機構の解明を目指したケミカルバイオロジー研究

    日本学術振興会  科学研究費助成事業 基盤研究(C)

    Project Year :

    2021.04
    -
    2024.03
     

    二村 友史

  • がん微小環境における代謝可塑性の理解と制御

    科学研究費助成事業  基盤研究(C)

    Project Year :

    2017.04
    -
    2020.03
     

    二村 友史

  • 形態異常を誘起する抗がん物質の創薬基盤研究

    科学研究費助成事業  若手研究(B)

    Project Year :

    2013.04
    -
    2015.03
     

    二村 友史

  • モルフォロームを基盤とした微生物由来新奇バイオプローブの探索研究

    科学研究費助成事業  若手研究(B)

    Project Year :

    2010.04
    -
    2012.03
     

    二村 友史

Presentations

  • Phenotypic screening, iHOPE and MorphoBase, for drug screening

    Yushi Futamura  [Invited]

    The 4th Chemical Biology Luncheon Seminar 

    Presentation date: 2021.11

  • 細胞形態を指標とした生理活性物質の探索

    二村 友史  [Invited]

    日本学術振興会第189委員会 2019年度第3回定例会 

    Presentation date: 2019.12

  • 細胞形態変化を利用した抗真菌物質の探索

    二村 友史, Rachael Uson-Lopez, 山本 甲斐, 室井 誠, 長田 裕之  [Invited]

    日本農芸化学会2019年度大会 

    Presentation date: 2019.03

  • 天然物創薬における苦しみと楽しみ

    二村 友史  [Invited]

    慶應義塾大学 特別講義 

    Presentation date: 2017.01

  • 細胞形態変化データベース「モルフォベース」の構築とその応用

    二村 友史  [Invited]

    GE Life Sciences Day 2013 

    Presentation date: 2013.07