Enterolignans (enterodiol and enterolactone) exhibit structural similarity to estradiol and have therefore been hypothesized to modulate hormone related cancers such as breast cancer. The bioactivation of the plant lignan secoisolariciresinol diglucoside (SDG) requires the transformation by intestinal bacteria including the deglycosylation of SDG to secoisolariciresinol (SECO) followed by demethylation and dehydroxylation of SECO to enterodiol (ED). Finally, ED is dehydrogenated to enterolactone (EL). It is unclear whether the bacterial activation of SDG to ED and EL is crucial for the cancer preventing effects of dietary lignans. The possible protective effect of bacterial lignan transformation on a 7,12 dimethylbenz(a)anthracene (DMBA)-induced breast cancer in gnotobiotic rats was investigated. Germ-free rats were associated with a defined lignan-converting consortium (Clostridium saccharogumia, Blautia producta, Eggerthella lenta, and Lactonifactor longoviformis). The rats colonized with lignan-converting bacteria consortium (LCC) were fed a lignan-rich flaxseed diet and breast cancer was chemical induced. Identically treated germ-free rats served as control. All bacteria of the consortium successfully colonized the intestine of the LCC rats. The plant lignan SDG was converted into the enterolignans ED and EL in the LCC rats but not in the germ-free rats. This transformation did not influence cancer incidence but significantly decreased tumor numbers per tumor-bearing rat, and tumor size. Cell proliferation was significantly inhibited and apoptosis was significantly induced in LCC rats. No differences between LCC and control rats were observed in the expression of the genes encoding the estrogen receptors (ERα and ERβ) and G-coupled protein receptor 30 (GPR30). Similar findings were observed for both insulin-like growth factor 1 (IGF-1) and epidermal growth factor receptor (EGFR) genes involved in tumor growth. Proteome analysis revealed that 24 proteins were differentially expressed in tumor tissue from LCC and germ-free. RanBP-type and C3HC4-type zinc finger-containing protein 1 (RBCK1) and poly(rC)-binding protein 1 (PBCP1) were down-regulated by 3.2- and 2.0-fold, respectively. These proteins are associated with cell proliferation. The activity of selected enzymes involved in the degradation of oxidants in plasma and liver was significantly increased in the LCC rats. However, plasma and liver concentrations of reduced glutathione (non-enzymatic antioxidant) and malondialdehyde (oxidative stress marker) did not differ between the groups. In conclusion, the bacterial conversion of plant lignan to enterolignans beneficially influences their anti-cancer effect. However, the mechanisms involved in these effects remain elusive. / Enterolignanen (Enterodiol ED und Enterolacton EL) wird aufgrund ihrer strukturellen Ähnlichkeit zu Estradiol ein modulierender Einfluss auf hormonell bedingte Krebserkrankungen wie Brustkrebs nachgesagt. Das pflanzliche Lignan Secoisolariciresinoldiglucosid (SDG) wird durch Darmbakterien zum Enterolignan aktiviert. Dies erfolgt über dessen Deglykosylierung zu Secoisolariciresinol (SECO) gefolgt durch die Demethylierung und die Dehydroxylierung zu Enterodiol (ED). Schließlich wird ED zu Enterolacton (EL) dehydrogeniert. Es ist allerdings noch nicht bewiesen, dass die bakterielle Aktivierung von SDG zu ED und EL für die antikanzerogenen Wirkungen verantwortlich ist, die für dieses in der menschlichen Ernährung vorkommende Lignan beschrieben wurden. Um dies zu klären, wurde der Einfluss der bakteriellen Lignan-Transformation auf die Protektion gegenüber einem durch 7,12-Dimethylbenz(a)anthracen (DMBA)-induzierten Brustkrebs im gnotobiotischen Rattenmodell untersucht. Keimfreie Ratten wurden hierfür mit einem Konsortium aus vier Bakterienstämmen (Clostridium saccharogumia, Blautia producta, Eggerthella lenta, und Lactonifactor longoviformis) besiedelt, das die Umsetzung von SDG zu ED und EL katalysiert (LCC-Ratten). Ratten, die über den gesamten Versuchszeitraum keimfrei blieben, dienten als Kontrolle. Die Tiere wurden über 16 Wochen mit einer Leinsamen-Diät gefüttert, die reich an pflanzlichen Lignanen war. Während der Fütterung wurde bei allen Tieren Brustkrebs chemisch induziert. Das pflanzliche Lignan SDG wurde nur in den LCC Ratten zu den Enterolignanen ED und EL umgewandelt. Keimfreie Ratten zeigten keine Transformation von SDG. Die bakterielle Transformation von SDG hatte zwar keinen Einfluss auf die Inzidenz von Brustkrebs, jedoch verringerten sich durch die Besiedlung der Ratten mit SDG-transformierenden Bakterien die Anzahl von Tumoren pro tumortragender Ratte und die Tumorgröße deutlich. Zudem wurde die Zellproliferation in den LCC-Ratten deutlich gehemmt und die Apoptose induziert. Unterschiede in der Genexpression der Östrogenrezeptoren (ERα und ERß) und G-Protein-gekoppelte Rezeptoren (GPR30) wurden zwischen den LCC-Ratten und den Kontrolltieren nicht beobachtet. Ebenso verhielt es sich für die Gene des Insulinähnliche Wachstumsfaktoren 1 (IGF-1) und Epidermale Wachstumsfaktor rezeptoren (EGFR), welche in das Tumorwachstum involviert sind. Die Analyse des Proteoms des Tumorgewebes ergab 24 differentiell exprimierte Proteine zwischen keimfreien und LCC-Ratten. So wurden zum Beispiel die Proteine RanBP-type and C3HC4-type zinc finger-containing protein 1 (RBCK1) und poly(rC)-binding protein 1 (PBCP1), die mit der Zellproliferation assoziiert sind, in LCC-Ratten um das 3,2 bzw. 2,0-fache herunterreguliert. Die Aktivität ausgewählter antioxidativer Enzyme in Plasma und Leber war in den LCC-Ratten im Vergleich zu den keimfreien Tieren deutlich erhöht. Allerdings unterschieden sich die Konzentrationen von reduziertem Glutathion (nichtenzymatisches Antioxidans) und Malondialdehyd (oxidativer Stress-Marker) in Plasma und Leber nicht zwischen den beiden Besiedlungs-Gruppen. Zusammenfassend zeigen die Ergebnisse, dass die bakterielle Umwandlung von pflanzlichen Lignanen zu Enterolignanen deren antikanzerogene Wirkung entscheidend beeinflusst. Allerdings bleiben die zugrunde liegenden Mechanismen weiterhin ungeklärt.

A sesquiterpene hydroquinone, Aureol (6), was isolated from extracts of Taiwanese Smenospongia sp. collected at Green Island in Taiwen. 6 was acylated to yield a series of derivatives and their structures were identified as O-Methylaureol (40)¡BAureol acetate (7)¡BAureol N,N-dimethylthiocarbamate (41)¡BO-Benzoyl-aureol (43)¡BO-4-Chlorobenzoyl-aureol (44)¡BO-4-Flurobenzoyl-aureol (45)¡BO-Nicotinoyl-aureol (46)¡BO-4-Methylbenzoyl-aureol (47) ¡BO-2-Thienylcarbonyl-aureol(48) ¡BO-2-Furoylcarbonyl-aureol (49) on the basis of 1D and 2D NMR techniques, including 1H-NMR¡B13C-NMR ¡BCOSY¡BHMQC¡BHMBC experiments. Preliminary pharmacological study revealed that some of the compounds possessed significant cytotoxicity against human cancer cells (Hepa59T/VGH¡BHela¡BKB). Kadsura philippinensis Elmer is medicinal plant which is rich in C18 type dibenzocyclooctadiene lignans. kadsuphilin A (50)¡B kadsuphilin B (51)¡B kadsuphilin C (52)¡B kadsuphilin D (53)¡B kadsurin (54)¡B kadsulignan D (55)¡Bkadsulignan K (56)¡Bbinankadsyrin A (57) ¡Bangeloyl-binankadsurin A (58) ¡Bkadsumarin A (59)¡Bschizanrin F (60) ¡Bschizanrin J (61) isolated from the lipophilic extract of aerial part of Kadsura philippinensis collected at Green Island in Taiwen. It is the first time to isolate Kadsuphilins A-D (50, 51, 52, 53). Kadsulignan D (55) and Kadsulignan K (56) were confirmed by single crystal X-ray analysis. The structures of new compounds were elucidated on the basis of 1D- and 2D-NMR techniques including COSY, HMQC, HMBC and NOESY experiments. Preliminary pharmacological study revealed that some of the compounds possessed significant cytotoxicity against human cancer cells.

bstract Cancer has been the first lethal factor in Taiwan, and hepatitis B is also a serious problem in our country. We have identified that the extracts of Taiwanese Schisandraceous plants, including Schizandra arisanensis, Kadsura matsudai and K. japonica, have inhibitory effect on type B hepatitis surface antigen (HBsAg) and e-antigen (HBeAg). Besides, we also find that the extracts of Echinops grijsii can inhibit the growth of human cancer cell line, KB and Hela. To purify active compounds by using column chromatography and high-performance liquid chromatography, we have furnished fifty-eight compounds from above Schisandraceous plants, including thirty-seven C18 lignans composed of [6.8.6]-dibenzocyclooctadiene skeleton, nine C19 homolignans, four triterpenes, three steroids and five cyclic-aromatic compounds. In the case of Echinops grijsii, we got thirteen major compounds containing eight thiophenes, four triterpenes, and a sterol. Structural elucidation of the novel homolignan and ten lignans are based on the spectral and chemical analyses, mainly by using two-dimensional nuclear magnetic resonance (NMR) of 1H and 13C nucleus. From the anti-HBsAg and anti-HBeAg assay, we found that four of new C18 lignans and one known (+)-gomisin K3 (33) lignan exhibited the significant inhibitory effects on surface antigen of hepatitis B virus. Moreover, to modify the known lignans, kadsurarin with different halogens and functional group having nitrogen atom, as well as (+)-gomisin K3 with several kinds of phenyl compounds and sulfur functional groups are processed. The preliminary structures and bioactivity relationships (SAR) studies demonstrated that (+)-gomisin K3 with sulfuric functional group could decrease cytotoxicity and increase inhibitory effect on surface antigen and e-antigen of hepatitis B virus. In the part of studying synthesis by using inactive 5¡¦-(but-3-en-1-ynyl)-2,2¡¦ -bithiophene (1) from Echinops grijsii, the yne-ene moiety of its structure was hydrolyzed into carbonyl group and then was attached a serious of various carbon number of hydroxy groups. Besides the above experiment, £\-trithienyl (1) was served as a substrate and made it into unsaturated carbonyl group, which then was linked with a series of hydroxy groups. Cytotoxicity datum showed that the derivatives of bi-thiophene have better anticancer activity than derivatives of tri-thiophene. And the assay results also exhibited that the bi-thiophene derivatives with hydroxy group with less than three carbon numbers have better inhibitory activity against cancer cells.

No description available.

Partindo-se de piperonal e succinato de metila, obteve-se o ácido 4-(3,4- metilenodioxifenil)-3-metoxicarbonil-3-butenóico (3), com rendimento de 60% e partindo-se deste, por redução catalítica obteve-se o ácido 4-(3,4- metilenodioxifenil)-3-metoxicarbonil-3-butanóico (4), com rendimento de 80%. A partir do (4), reagindo-se com Ca(BH4)2 e H+, obteve-se a 4-(3,4- metilenodioxifenil)-4,5-di-hidro-2(3H)-furanona (5), com rendimento de 70 %. Pela adição de piperonal, sintetizou-se a 7-hidroxi-hinoquinina (6), com rendimento de 89% da mistura dos diasteroisômeros (6a e 6b), os quais foram separados por cristalização. A reação de ambos os compostos (6a e 6b) com CF3CO forneceu o derivado ariltetralínico (7), com 98% de rendimento. A poligamaina (7) foi reduzida com DIBAL-H em THF, fornecendo o composto (9). O derivado (9), constituído de uma mistura enanciomérica foi separado em CLAE quiral. Os derivados foram analisados por RMN 1H, BB e DEPT. Com relação aos ensaios biológicos, os compostos 6, 7 e 9 foram avaliados patógenos bucais para determinação dos valores de CIM. Os melhores resultados obtidos para o composto (+) 9 foram contra S. mutans (250 µM), S. salivarius (250 µM), S. sobrinus (280 µM) e S. mitis (280 µM). O isômero (-) 9 foi ativo contra S. sanguinis (280 µM) enquanto (9) mostrou melhor atividade contra L. casei (370 µM) e E. faecalis (710 µM). No ensaio tripanocida, pode-se observar que o melhor resultado foi obtido para a mistura de enanciômeros (9), a qual apresentou IC50 = 1,4 µM e lise de 61,9% ± 0,9 na concentração de 32 µM. Por outro lado, ambos os enanciômeros isolados foram menos ativos apresentando IC50 de 351,8 µM e 135 µM para (-) 9 e (+) 9, respectivamente e lise de 47,3% ± 5,9 na concentração de 128 µM, para o enanciômero (+). / The (3,4-methylenodioxiphenyl)-3-methoxycarbonyl-3-butenoic acid (3) was obtained in 60 % yield by reacting pyperonal and methyl succinate. Acid 3 was submitted to catalytic reduction furnishing the 4-(3,4-methylenedioxiphenyl)-3- methoxicarbonyl-3-butanoic acid (4), yielding 80 %. Then, compound 4 was reacted with Ca(BH4)2 and H+, furnishing 4-(3,4-methylenedioxiphenyl)-4,5-dihydro-2(3H)-furanone (5), yielding 70 %. Piperonal was added to compound 5 to produce 7-hidroxyhinoquinin (6), yielding 89% of a mixture of diasteroisomers (6a and 6b), which was separate by crystallization. The reaction of both compounds 6a e 6b with CF3CO furnished an aryltetralin lignan derivative poligamain (7), yielding 98 %. Poligamain was reduced by using DIBAL-H in THF, to furnish compound 9. Compound 9, constituted of an enantiomeric mixture was separated by chiral HPLC. All the obtained compounds were analyzed by 1H NMR, BB and DEPT techniques. Regarding the biological assays, the compounds 6, 7 and 9 were assayed against oral pathogens by determining its MIC values. The best results obtained for the (+) 9 isomer was against S. mutans (250 µM), S. salivarius (250 µM), S. sobrinus (280 µM) and S. mitis (280 µM). For the (-) 9 isomer it was active against S. sanguinis (280 µM) while (9) displayed higher activity against L. Casei (370 µM) and E. faecalis (710 µM). Regarding the in vitro trypanocidal assay, on one hand the best result was observed for the mixture of enantiomers (9), which displayed IC50 = 1.4 µM and lysis of 61.9% ± 0.9 at 32 µM. On the other hand, both isolated enantiomers were less active by displaying IC50 of 351.8 µM and 135 µM for (-) 9 and (+) 9, respectively and lysis of 47.3% ± 5.9 at 128 µM, for (+) enantiomer.

The goal of this PhD research was directed to the isolation and structural elucidation of the secondary metabolites from four species of Durio plants, especially from those growing in the island of Borneo; This island contains most Durio species. The structures of isolated compounds were deduced using state-of-the-art approaches, including 1D and 2D nuclear magnetic resonance (NMR), mass spectrometric (MS) and circular dichroism (CD) analyses. From the bark of the woody species Durio zibethinus Murr. (Bombacaceae), that was collected in Pontianak, a chemically diverse group of oxygenated compounds was obtained. These included caffeoyl triterpenes, lignans and phenylpropanoid derivatives. During this research, one new compound 27-O-cis-caffeoylcylicodiscate was obtained which resulted from a trans to cis isomerization; this process might be caused by acid and light. Two presumed fungal metabolites were also isolated during this study, these possibly arising from fungal (unknown species) contamination of the bark. The relative stereochemistry of 4-hydroxymellein were also established by conversion to it diacetate and measurement of the coupling constant between H-3 and H-4. The absolute configuration was determined by comparison of []D values with those from the literature. Assignment of the 13C NMR spectrum for (+)-(R)-de-O-methyllasiodiplodin was completed using HSQC-TOCSY data. The methanol extract of the bark of Durio kutejensis (Hassk.) Becc. was fractionated and purified by C18-HPLC. Two new caffeoyl pentacyclic triterpenes 3-O-trans-caffeoyl-2-hydroxyolean-12-en-28-oic acid and 3-O-trans-caffeoyl-2-hydroxyurs-12-en-28-oic acid, together with five known compounds were isolated. The known compound fraxidin had previously been obtained from D. zibethinus Murr. The bark of Durio carinatus Mast., a plant species which has non edible fruit was collected in Sambas district, West Kalimantan. Three new lignans, boehmenan X, (-)-(7S,8S)-threo-carolignan X and (-)-(7R,8S)-erythro-carolignan X, and a new caffeoyl triterpene 3-O-cis-caffeoyl betulinic acid, together with three known lignans and one known caffeoyl triterpene, were obtained from the crude MeOH extract. The relative configurations and conformational models of the diastereomers threo- and erythro-carolignan X were determined by the coupling constant data for H-7′ and H-8′ and supported by 2D NMR spectroscopic analysis including NOESY and HSQC-HECADE in CDCl3. However, the magnitude of the coupling constants for both compounds in MeOH-d4 was similar. The stability of the individual conformers was solvent dependent and must be considered for their contribution to the overall conformational equilibria. This stability was determined by steric interactions, dipole repulsion effects and intramolecular hydrogen bonding. The absolute configuration of the two new diastereomers was established by application of circular dichroism (CD) and optical rotation []D measurements. Another Durio plant studied and collected in Bengkayang region, West Kalimantan, was Durio oxleyanus Griff. From the MeOH extract of its bark, two new lignans, (-)-(7S,8S)-threo-carolignan Y and (-)-(7R,8S)-erythro-carolignan Y were isolated using C18-HPLC. The relative stereochemistry of the carolignan Y diastereomers could not be deduced from the coupling constants for H-7′ and H-8′ either in CDCl3 and MeOH-d4, because of the presence of multiple conformers. The conformational model for the diastereomers of carolignan Y was similar to that of threo- and erythro-carolignan X in MeOH-d4. Using 2D NMR spectroscopic methods, including NOESY and HSQC-HECADE, the relative stereochemistry of the threo and erythro isomers of carolignan Y was confirmed. Further, the absolute configuration of threo- and erythro-carolignan Y was explored via analysis of CD spectra and by comparison of experimentally obtained []D values with literature values.

Partindo-se de piperonal e succinato de metila, obteve-se o ácido 4-(3,4- metilenodioxifenil)-3-metoxicarbonil-3-butenóico (3), com rendimento de 60% e partindo-se deste, por redução catalítica obteve-se o ácido 4-(3,4- metilenodioxifenil)-3-metoxicarbonil-3-butanóico (4), com rendimento de 80%. A partir do (4), reagindo-se com Ca(BH4)2 e H+, obteve-se a 4-(3,4- metilenodioxifenil)-4,5-di-hidro-2(3H)-furanona (5), com rendimento de 70 %. Pela adição de piperonal, sintetizou-se a 7-hidroxi-hinoquinina (6), com rendimento de 89% da mistura dos diasteroisômeros (6a e 6b), os quais foram separados por cristalização. A reação de ambos os compostos (6a e 6b) com CF3CO forneceu o derivado ariltetralínico (7), com 98% de rendimento. A poligamaina (7) foi reduzida com DIBAL-H em THF, fornecendo o composto (9). O derivado (9), constituído de uma mistura enanciomérica foi separado em CLAE quiral. Os derivados foram analisados por RMN 1H, BB e DEPT. Com relação aos ensaios biológicos, os compostos 6, 7 e 9 foram avaliados patógenos bucais para determinação dos valores de CIM. Os melhores resultados obtidos para o composto (+) 9 foram contra S. mutans (250 µM), S. salivarius (250 µM), S. sobrinus (280 µM) e S. mitis (280 µM). O isômero (-) 9 foi ativo contra S. sanguinis (280 µM) enquanto (9) mostrou melhor atividade contra L. casei (370 µM) e E. faecalis (710 µM). No ensaio tripanocida, pode-se observar que o melhor resultado foi obtido para a mistura de enanciômeros (9), a qual apresentou IC50 = 1,4 µM e lise de 61,9% ± 0,9 na concentração de 32 µM. Por outro lado, ambos os enanciômeros isolados foram menos ativos apresentando IC50 de 351,8 µM e 135 µM para (-) 9 e (+) 9, respectivamente e lise de 47,3% ± 5,9 na concentração de 128 µM, para o enanciômero (+). / The (3,4-methylenodioxiphenyl)-3-methoxycarbonyl-3-butenoic acid (3) was obtained in 60 % yield by reacting pyperonal and methyl succinate. Acid 3 was submitted to catalytic reduction furnishing the 4-(3,4-methylenedioxiphenyl)-3- methoxicarbonyl-3-butanoic acid (4), yielding 80 %. Then, compound 4 was reacted with Ca(BH4)2 and H+, furnishing 4-(3,4-methylenedioxiphenyl)-4,5-dihydro-2(3H)-furanone (5), yielding 70 %. Piperonal was added to compound 5 to produce 7-hidroxyhinoquinin (6), yielding 89% of a mixture of diasteroisomers (6a and 6b), which was separate by crystallization. The reaction of both compounds 6a e 6b with CF3CO furnished an aryltetralin lignan derivative poligamain (7), yielding 98 %. Poligamain was reduced by using DIBAL-H in THF, to furnish compound 9. Compound 9, constituted of an enantiomeric mixture was separated by chiral HPLC. All the obtained compounds were analyzed by 1H NMR, BB and DEPT techniques. Regarding the biological assays, the compounds 6, 7 and 9 were assayed against oral pathogens by determining its MIC values. The best results obtained for the (+) 9 isomer was against S. mutans (250 µM), S. salivarius (250 µM), S. sobrinus (280 µM) and S. mitis (280 µM). For the (-) 9 isomer it was active against S. sanguinis (280 µM) while (9) displayed higher activity against L. Casei (370 µM) and E. faecalis (710 µM). Regarding the in vitro trypanocidal assay, on one hand the best result was observed for the mixture of enantiomers (9), which displayed IC50 = 1.4 µM and lysis of 61.9% ± 0.9 at 32 µM. On the other hand, both isolated enantiomers were less active by displaying IC50 of 351.8 µM and 135 µM for (-) 9 and (+) 9, respectively and lysis of 47.3% ± 5.9 at 128 µM, for (+) enantiomer.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第18336号 / 農博第2061号 / 新制||農||1023(附属図書館) / 学位論文||H26||N4843(農学部図書室) / 31194 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 梅澤 俊明, 教授 矢﨑 一史, 教授 三上 文三 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

No description available.

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