論文 - 荒井　緑

beta-Sitosterol and flavonoids isolated from Bauhinia malabarica found during screening for Wnt signaling inhibitory activity

Hyun Young Park, Kazufumi Toume, Midori A. Arai, Takashi Koyano, Thaworn Kowithayakorn, Masami Ishibashi

JOURNAL OF NATURAL MEDICINES （SPRINGER JAPAN KK）  68 （ 1 ） 242 - 245 2014年01月

査読有り,  ISSN  1340-3443

　概要を見る

Screening with a cell-based luciferase assay was conducted to identify bioactive natural products which inhibit Wnt signaling activity-guided separation of an MeOH extract of Bauhinia malabarica (Caesalpiniaceae) leaves yielded five compounds, which were identified as beta-sitosterol (1), quercetin (2), 6,8-C-dimethyl kaempferol-3-O-rhamnopyranoside (3), hyperin (4), and 6,8-C-dimethyl kaempferol-3-methyl ether (5). The tested compounds 1, 3, and 5 exhibited Wnt signaling inhibitory activity, with IC50 values of 0.77, 0.74, and 16.6 mu M, respectively.

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CONSTITUENTS FROM THE RHIZOMES OF CURCUMA COMOSA AND THEIR WNT SIGNAL INHIBITORY ACTIVITIES

Rolly G. Fuentes, Kazufumi Toume, Midori A. Arai, Takashi Koyano, Thaworn Kowithayakorn, Masami Ishibashi

HETEROCYCLES （PERGAMON-ELSEVIER SCIENCE LTD）  88 （ 2 ） 1501 - 1509 2014年01月

査読有り,  ISSN  0385-5414

　概要を見る

Activity-guided fractionation of the Curcuma comosa rhizome MeOH extract that inhibited Wnt signal activity led to the characterization of two new diarylheptanoids (1 and 2) together with six known diarylheptanoids (3-8) and two known sesquiterpenoids (9 and 10). Their structures were identified by 1D and 2D NMR, HRESIMS, optical rotation, and chemical evidences. Compounds 1 and 8 exhibited dose-dependent inhibitory activity against beta-catenin/TCF transcriptional activity.

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Physalin H from Solanum nigrum as an Hh signaling inhibitor blocks GLI1-DNA-complex formation

Midori A. Arai, Kyoko Uchida, Samir K. Sadhu, Firoj Ahmed, Masami Ishibashi

BEILSTEIN JOURNAL OF ORGANIC CHEMISTRY （BEILSTEIN-INSTITUT）  10   134 - 140 2014年01月

査読有り,  ISSN  1860-5397

　概要を見る

Hedgehog (Hh) signaling plays an important role in embryonic development, cell maintenance and cell proliferation. Moreover, Hh signaling contributes to the growth of cancer cells. Physalins are highly oxidized natural products with a complex structure. Physalins (1-7) were isolated from Solanum nigrum (Solanaceae) collected in Bangladesh by using our cell-based assay. The isolated physalins included the previously reported Hh inhibitors 5 and 6. Compounds 1 and 4 showed strong inhibition of GLI1 transcriptional activity, and exhibited cytotoxicity against cancer cell lines with an aberrant activation of Hh signaling. Compound 1 inhibited the production of the Hh-related proteins patched (PTCH) and BCL2. Analysis of the structures of different physalins showed that the left part of the physalins was important for Hh inhibitory activity. Interestingly, physalin H (1) disrupted GLI1 binding to its DNA binding domain, while the weak inhibitor physalin G (2) did not show inhibition of GLI1-DNA complex formation.

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Calotropin, a cardenolide from Calotropis gigantea, with Wnt signal inhibition activity through increase of casein kinase 1 alpha

Park, H. Y., Toume, K., Arai, M. A., Sadhu, S. K., Ahmed, F., Ishibashi, M.

Planta Medica 80 （ 10 ） 818 - 818 2014年

査読有り

New aromatic heterocycles with TRAIL resistance overcoming activity isolated from actinomycetes collected in Chiba area

Abdelfattah, M. S., Toume, K., Arai, M. A., Ishibashi, M.

Planta Medica 80 （ 10 ）  2014年

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Prenylflavonoids isolated from Artocarpus champeden with TRAIL-resistance overcoming activity

Tomohiro Minakawa, Kazufumi Toume, Midori A. Arai, Takashi Koyano, Thaworn Kowithayakorn, Masami Ishibashi

PHYTOCHEMISTRY （PERGAMON-ELSEVIER SCIENCE LTD）  96   299 - 304 2013年12月

査読有り,  ISSN  0031-9422

　概要を見る

In a screening program for bioactive natural products which can overcome Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-resistance, three prenylflavonoids, named pannokin A-C, were isolated from a MeOH extract of Artocarpus champeden (Moraceae) roots, together with three known prenylflavonoids. The structures of pannokin A-C were elucidated by spectroscopic analysis. These of the prenylflavonoids in combination with TRAIL, showed cytotoxic activity in sensitizing TRAIL-resistant human gastric adenocarcinoma (AGS) cells. Of these compounds, heterophyllin increased caspase 3/7 activity when combined with TRAIL in AGS cells, and enhanced the expression of DR4 and DR5 mRNA. Moreover, heterophyllin up-regulated mRNA expression of CCAAT/enhancer-binding protein-homologous protein (CHOP) which was reported to be an important regulator of DR5 expression. Thus, heterophyllin was presumed to cause a CHOP-dependent up-regulation of DR5 expression resulting in apoptosis in AGS cells. (C) 2013 Elsevier Ltd. All rights reserved.

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Xylogranin B: A Potent Wnt Signal Inhibitory Limonoid from Xylocarpus granatum

Toume, Kazufumi, Kamiya, Kentaro, Arai, Midori A., Mori, Naomi, Sadhu, Samir K., Ahmed, Firoj, Ishibashi, Masami

ORGANIC LETTERS （AMER CHEMICAL SOC）  15 （ 23 ） 6106 - 6109 2013年12月

査読有り,  ISSN  1523-7060

　概要を見る

Xylogranin B (2) was isolated from Xylocarpus granatum (Meliaceae) leaves, by use of a cell-based luciferase screening system targeting a Wnt signaling pathway. Compound 2 inhibited TCF/beta-catenin transcriptional activity (IC50 48.9 nM) and exhibited strong cytotoxicity against colon cancer cell lines. Compound 2 significantly decreased beta-catenin protein levels in nuclei but not in the cytosol. These results indicated that a decrease in beta-catenin levels in nuclei by 2 resulted in the Wnt signal inhibitory effects of 2.

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Hh signaling inhibitors from Vitex negundo; Naturally occurring inhibitors of the GLI1-DNA complex

Midori A. Arai, Teruhisa Fujimatsu, Kyoko Uchida, Samir K. Sadhu, Firoj Ahmed, Masami Ishibashi

Molecular BioSystems 9 （ 5 ） 1012 - 1018 2013年05月

査読有り,  ISSN  1742-206X

　概要を見る

The hedgehog (Hh) signaling pathway has crucial roles in embryonic development, cell maintenance and proliferation, and is also known to contribute to cancer cell growth. New naturally occurring Hh inhibitors (1, 7 and 9) were isolated from Vitex negundo using our previously constructed cell-based assay. Bioactivity guided isolation provided 9 natural compounds including a new diterpene, nishindanol (9). Compounds 7 and 9 showed cytotoxicity against cancer cell lines in which Hh signaling was aberrantly activated. Vitetrifolin D (7
GLI1 transcriptional inhibition IC50 = 20.2 μM) showed inhibition of Hh related protein (PTCH and BCL2) production. Interestingly, the constructed electrophoresis mobility shift assay revealed that vitetrifolin D (7) disrupted GLI1 binding on its DNA binding domain. epi-Sclareol (8
inactive), possessing a similar structure to 7, did not show inhibition of GLI1-DNA complex formation. This is the first example of naturally occurring inhibitors of GLI1-DNA complex formation. © 2013 The Royal Society of Chemistry.

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Merging Chemistry and Biology in Emerging Countries

Takayoshi Suzuki, Midori A. Arai, Mitsue Nakashima, Motonari Uesugi

CHEMISTRY & BIOLOGY （CELL PRESS）  20 （ 4 ） 461 - 465 2013年04月

査読有り,  ISSN  1074-5521

　概要を見る

Chemical biology is not a new discipline, as research at the interface of chemistry and biology has been growing for decades. However, the field still needs regional and global support in emerging countries. The meeting organized by Asian Chemical Biology Initiative in Bangkok, Thailand in January 2013, brought together regional communities of young and established researchers to nurture chemical biology research programs and highlight some of the newer developments. Here, we report on the meeting and some of the key research topics discussed.

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Catalytic Asymmetric Synthesis of Mixed 3,3 '-Bisindoles and Their Evaluation as Wnt Signaling Inhibitors

Takayoshi Arai, Yushi Yamamoto, Atsuko Awata, Kentaro Kamiya, Masami Ishibashi, Midori A. Arai

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION （WILEY-V C H VERLAG GMBH）  52 （ 9 ） 2486 - 2490 2013年

査読有り,  ISSN  1433-7851

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Naturally occurring Ngn2 promoter activators from Butea superba

Midori A. Arai, Kazune Koryudzu, Takashi Koyano, Thaworn Kowithayakorn, Masami Ishibashi

MOLECULAR BIOSYSTEMS （ROYAL SOC CHEMISTRY）  9 （ 10 ） 2489 - 2497 2013年

査読有り,  ISSN  1742-206X

　概要を見る

Neurogenin2 (Ngn2), an activator-type bHLH transcriptional factor, promotes differentiation of neural stem cells into neurons by transcription of pro-neural genes. To find neural stem cell accelerators from the extract library of natural resources, we used a two-step screening including a Ngn2 promoter reporter gene screening and differentiation assay screening of neural stem cells. A reporter gene assay that can detect Ngn2 promoter activity by luciferase expression was constructed using C3H10T1/2 cells. Using this primary cell-based screening, Butea superba was found to include Ngn2 promoter activators from our tropical plant extract libraries. Bioassay-guided fractionation of this plant extract led to the isolation of 18 natural products, including pterocarpans and isoflavonoids. Dehydromaackiain (1), formononetin (6), (-)-variabilin (13), (-)-medicarpin (14), rothindin (17) and ononin (18) showed 1.8-2.8 times higher Ngn2 promoter activity at 5 mu M compared with control. Of active natural compounds, 3'-methoxydaidzein (3) showed promotion of neurite outgrowth of C17.2 in a secondary screen. 3'-Methoxydaidzein (3) increased mRNA expression of pro-neural transcriptional factors (Ngn2, Ngn1, NeuroD2), a mature neuron-specific enzyme GAD1 and a pro-neural neurotrophic growth factor neurotrophin 3 (NT3) in C17.2 neural stem cells.

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GRISEOVIRIDIN AND CYCLIC HYDROXAMATES FOUND IN A SCREENING PROGRAM FOR WNT SIGNAL INHIBITOR

Tamai, Yuuya, Toume, Kazufumi, Arai, Midori A., Ishibashi, Masami

HETEROCYCLES （PERGAMON-ELSEVIER SCIENCE LTD）  86 （ 2 ） 1517 - 1524 2012年12月

査読有り,  ISSN  0385-5414

　概要を見る

A screening study using a luciferase assay to identify natural products which inhibit Wnt signaling was carried out. Bioassay-guided fractionation of the culture broth and mycelia of actinomycetes CKK748 and CKK784, led to the isolation of griseoviridin (1), and four cyclic hydroxamates (4-7) as active compounds. 1, 4, and 5 inhibited TCF/beta-catenin transcriptional activity with IC50 values of 8.2-14 mu M.

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Cycloartane Triterpenes and Ingol Diterpenes Isolated from Euphorbia neriifolia in a Screening Program for Death-Receptor Expression-Enhancing Activity

Toume, Kazufumi, Nakazawa, Takafumi, Hoque, Tahmina, Ohtsuki, Takashi, Arai, Midori A., Koyano, Takashi, Kowithayakorn, Thaworn, Ishibashi, Masami

PLANTA MEDICA （GEORG THIEME VERLAG KG）  78 （ 12 ） 1370 - 1377 2012年08月

査読有り,  ISSN  0032-0943

　概要を見る

In our screening program for natural products that increase death-receptor 5 expression, seven new cycloartane triterpenes, euphonerins A-G (1-7), and 3-O-acetyl-8-O-tigloylingol (8), a new ingol diterpene, were isolated from the MeOH extract of Euphorbia neriifolia leaves, together with 3,12-di-O-acetyl-8-O-tigloylingol (9), (24R)-cycloartane-3 beta,24,25-triol (10), and three known flavonols (11-13). The structures of 1-8 were elucidated by spectroscopic analysis. Among these compounds, 1-11 showed death-receptor 5 expression enhancing activity.

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Eudesmane-Type Sesquiterpenoid and Guaianolides from Kandelia candel in a Screening Program for Compounds to Overcome TRAIL Resistance

Tomohiro Minakawa, Kazufumi Toume, Midori A. Arai, Samir K. Sadhu, Firoj Ahmed, Masami Ishibashi

JOURNAL OF NATURAL PRODUCTS （AMER CHEMICAL SOC）  75 （ 8 ） 1431 - 1435 2012年08月

査読有り,  ISSN  0163-3864

　概要を見る

In a screening program for natural products that can overcome TRAIL resistance, a new eudesmane-type sesquiterpenoid (1), three new guaianolides, mehirugins A-C (2-4), and two known guaianolides (5 and 6) were isolated from a Me OH extract of Kandelia candel leaves. Compounds 1 and 3-6 in combination with TRAIL showed cytotoxic activity in sensitizing TRAIL resistant human gastric adenocarcinoma cells.

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BIOACTIVE NATURAL PRODUCTS FROM MYXOMYCETES HAVING EFFECTS ON SIGNALING PATHWAYS

Masami Ishibashi, Midori A. Arai

HETEROCYCLES （PERGAMON-ELSEVIER SCIENCE LTD）  85 （ 6 ） 1299 - 1332 2012年06月

査読有り,  ISSN  0385-5414

　概要を見る

The myxomycetes (true slime molds) are an unusual group of primitive organisms that may be assigned to one of the lowest classes of eukaryotes. We studied the chemical constituents of a laboratory culture of plasmodia as well as field-collected fruit bodies of myxomycetes, and isolated various compounds with unique chemical structures and bioactivities. We recently also performed screening studies of natural products targeting signaling pathways related to basic studies of diseases and life sciences. Among myxomycetes constituents, we found several bioactive compounds that have effects on signaling pathways, including a cycloanthranilylproline with TRAIL resistance-overcoming activity, peptide lactone derivatives with Wnt signal inhibitory activity, a naphthoquinone pigment with Hes1 dimerization inhibition activity, and bisindole alkaloids with hedgehog signal inhibitory activity. Here we describe our recent results of their isolation, structure elucidation, synthesis, and findings in bioactivity studies.

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Katorazone, a new yellow pigment with a 2-azaquinone-phenylhydrazone structure produced by Streptomyces sp IFM 11299

Mohamed S. Abdelfattah, Kazufumi Toume, Midori A. Arai, Hyuma Masu, Masami Ishibashi

TETRAHEDRON LETTERS （PERGAMON-ELSEVIER SCIENCE LTD）  53 （ 26 ） 3346 - 3348 2012年06月

査読有り,  ISSN  0040-4039

　概要を見る

An unusual alkaloid with a 2-azaquinone-phenylhydrazone structure, katorazone (1), and other metabolites were isolated from the ethyl acetate extract of Streptomyces sp. IFM 11299. The chemical structure of katorazone (1) was elucidated by 1D and 2D NMR analyses together with HR-ESI mass spectrometry. Katorazone (1) showed a synergistic effect in combination with TRAIL and decreased the viability of human gastric adenocarcinoma (AGS) cells. (C) 2012 Elsevier Ltd. All rights reserved.

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Phorbol Esters with Wnt Signal-Augmenting Effects Isolated from Excoecaria indica

Yamaguchi, Tatsuhiro, Toume, Kazufumi, Arai, Midori A., Ahmed, Firoj, Sadhu, Samir K., Ishibashi, Masami

NATURAL PRODUCT COMMUNICATIONS （SAGE PUBLICATIONS INC）  7 （ 4 ） 475 - 477 2012年04月

査読有り,  ISSN  1934-578X

　概要を見る

A screening of natural products using a luciferase assay targeting the Wnt signaling pathway was carried out, and the bioassay-guided fractionation of Excoecaria indica (Euphorbiaceae) collected from Bangladesh afforded three phorbol esters (1 3). These compounds exhibited Wnt signal-augmenting effects with 1 causing a 25-fold increase in TCF/beta-catenin (TOP) transcriptional activity at 95 nM.

A New Resin Glycoside from Ipomoea maxima

Kazune Koryudzu, Midori A. Arai, Firoj Ahmed, Samir K. Sadhu, Masami Ishibashi

NATURAL PRODUCT COMMUNICATIONS （NATURAL PRODUCTS INC）  7 （ 2 ） 219 - 220 2012年02月

査読有り,  ISSN  1934-578X

　概要を見る

A new resin glycoside (1) was isolated from the aerial part of Ipomoea maxima, together with three known compounds, pescaprein XX (2), stoloniferin X (3), and stoloniferin IX (4). The structure of 1 was elucidated on the basis of ID NMR spectroscopy, a fragmentation study by APCIMS, and HRESIMS analysis.

NONACTIN AND RELATED COMPOUNDS FOUND IN A SCREENING PROGRAM FOR WNT SIGNAL INHIBITORY ACTIVITY

Yuuya Tamai, Kazufumi Toume, Midori A. Arai, Akinori Hayashida, Hikaru Kato, Yoshikazu Shizuri, Sachiko Tsukamoto, Masami Ishibashi

HETEROCYCLES （PERGAMON-ELSEVIER SCIENCE LTD）  84 （ 2 ） 1245 - 1250 2012年01月

査読有り,  ISSN  0385-5414

　概要を見る

A screening study using a luciferase assay to identify natural products with inhibitory activity against Wnt signaling was carried out. Fractionation of the culture extract of an actinomycete, strain CKK179, led to the identification of nonactin (1), monactin (2), and dinactin (3) as active compounds. We examined the effects of dimeric dinactin (4), isolated from another actinomycete, Streptomyces sp. (YM09-028), on Wnt signaling activity and found that 1 - 4 inhibited TCF/beta-catenin transcriptional activity with IC50 values of 0.6 - 7.4 nM.

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Mechanism of the anti-proliferative action of 25-hydroxy-19-nor-vitamin D-3 in human prostate cells

Eiji Munetsuna, Sachie Nakabayashi, Rie Kawanami, Kaori Yasuda, Miho Ohta, Midori A. Arai, Atsushi Kittaka, Tai C. Chen, Masaki Kamakura, Shinichi Ikushiro, Toshiyuki Sakaki

JOURNAL OF MOLECULAR ENDOCRINOLOGY （BIOSCIENTIFICA LTD）  47 （ 2 ） 209 - 218 2011年10月

査読有り,  ISSN  0952-5041

　概要を見る

According to the prevailing paradigm, 1 alpha-hydroxylation of 25-hydroxyvitamin D-3 (25(OH)D-3) and its analogs is a prerequisite step for their biological effects. We previously reported that 25-hydroxy-19-nor-vitamin D-3 (25(OH)-19-nor-D-3) had anti-proliferative activity in a cell line, PZ-HPV-7, which was derived from human non-cancerous prostate tissue, and suggested that 25(OH)-19-nor-D-3 acted after 1 alpha-hydroxylation by vitamin D 1 alpha-hydroxylase (CYP27B1). However, metabolic studies of 25(OH)-19-nor-D-3 using recombinant CYP27B1 revealed that 25(OH)-19-nor-D-3 was rarely subjected to 1 alpha-hydroxylation. Therefore, in this report, we attempted to clarify the mechanism of 25(OH)-19-nor-D-3 action in intact cells using PZ-HPV-7 prostate cells. After incubating the cells with 25(OH)-19-nor-D-3, eight metabolites of 24-hydroxylase (CYP24A1) were detected, whereas no products of CYP27B1 including 1 alpha,25-dihydroxy-19-nor-vitamin D-3 (1 alpha,25(OH)(2)-19-nor-D-3) were found. Furthermore, the time-dependent nuclear translocation of vitamin D receptor (VDR) and the subsequent transactivation of cyp24A1 gene in the presence of 25(OH)-19-nor-D-3 were almost identical as those induced by 1 alpha,25(OH)(2)-19-nor-D-3. These results strongly suggest that 25(OH)-19-nor-D-3 directly binds to VDR as a ligand and transports VDR into the nucleus to induce transcription of cyp24A1 gene. In addition, knock down of cyp27B1 gene did not affect the anti-proliferative activity of 25(OH)-19-nor-D-3, whereas knock down of VDR attenuated the inhibitory effect. Thus, our results clearly demonstrate that the anti-proliferative activity of 25(OH)-19-nor-D-3 is VDR dependent but 1 alpha-hydroxylation independent, suggesting that 25(OH)D-3 analogs such as 25(OH)-19-nor-D-3 could be attractive candidates for anticancer therapy. Journal of Molecular Endocrinology (2011) 47, 209-218

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