• Title/Summary/Keyword: IC-MS/MS

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Inhibitory Effect of Astragali Radix on Matrix Degradation in Human Articular Cartilage

  • CHOI SOOIM;PARK SO-RA;HEO TAE-RYEON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1258-1266
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    • 2005
  • The present study was carried out in order to assess the protective effects of calycosin-7-O-$\beta$-D-glucopyranoside, isolated from Astragali radix (AR), on hyaluronidase (HAase) and the recombinant human interleukin-$1\beta$ (IL-$1\beta$)-induced matrix degradation in human articular cartilage and chondrocytes. We isolated the active component from the n-butanol soluble fraction of AR (ARBu) as the HAase inhibitor and structurally identified as calycosin-7-O-$\beta$-D-glucopyranoside by LC-MS, IR, ${1}^H$ NMR, and ${13}^C$ NMR analyses. The $IC_{50}$ of this component on HAase was found to be 3.7 mg/ml by in vitro agarose plate assay. The protective effect of ARBu on the matrix gene expression of immortalized chondrocyte cell line C28/I2 treated with HAase was investigated using a reverse transcription polymerase chain reaction (RT-PCR), and its effect on HAase and IL-$1\beta$-induced matrix degradation in human articular cartilage was determined by a staining method and calculating the amount of degraded glycosaminoglycan (GAG) from the cultured media. Pretreatment with calycosin-7-O-$\beta$-D-glucopyranoside effectively protected human chondrocytes and articular cartilage from matrix degradation. Therefore, calycosin-7-O-$\beta$-D-glucopyranoside from AR appears to be a potential natural ant-inflammatory or antii-osteoarthritis agent and can be effectively used to protect from proteoglycan (PG) degradation.

Studies on Xanthine Oxidase Inhibitor Produced from Aspergillus sp. F184 (Aspergillus sp. F184가 생산하는 Xanthine Oxidase 저해제에 관한 연구)

  • 박시형;윤상웅;박정민;옥승호;유주현;배동훈
    • Microbiology and Biotechnology Letters
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    • v.28 no.2
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    • pp.92-96
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    • 2000
  • Aspergillus sp. F184 was isolated from soil for the development of new xanthine oxidase inhibitor. This xanthine oxidase inhibitor was sequentially purified by filtration, HP-20 adsorption column chromatography, ethyl acetate extraction, silica gel column chromatography and crystallization, and was named as YUX 104. YUX 104 was identified to be 5,6-epoxy-2-hydroxy-3-methyl-2-cyclohexene-1,4-dione(terreic acid) by NMR and mass spectroscopic sudies.

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Clitocybin D, a Novel Human Neutrophil Elastase Inhibitor from the Culture Broth of Clitocybe aurantiaca

  • Kim, Young-Hee;Ryoo, In-Ja;Choo, Soo-Jin;Xu, Guang-Hua;Lee, Sang-Ku;Seok, Soon-Ja;Bae, Ki-Hwan;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1139-1141
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    • 2009
  • Clitocybin D, a novel human neutrophil elastase inhibitor, was isolated from the culture broth of Clitocybe aurantiaca. This compound was purified by solvent extraction, silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative HPLC. The compound was determined to be 4-(4,6-dihydroxy-3-methoxy-3H-isoindol-1-yl)-benzoic acid on the basis of 1D and 2D NMRs and MS spectroscopic analysis. Analysis of the human neutrophil elastase (HNE) inhibitory activity of the isolated compound revealed that it showed significant HNE inhibitory activity with an $IC_{50}$ value of $17.8{\mu}M$.

Hydroxyhibiscone A, a Novel Human Neutrophil Elastase Inhibitor from Hibiscus syriacus

  • Ryoo, In-Ja;Yun, Bong-Sik;Lee, In-Kyoung;Kim, Young-Hee;Lee, Ik-Soo;Ahn, Jong-Seok;Bae, Ki-Hwan;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1189-1191
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    • 2010
  • In an ongoing investigation of compounds from natural products that exhibit anti-aging properties, hydroxyhibiscone A (1), a new furanosesquiterpenoid, together with hibiscone D (2), was isolated from the root bark of Hibiscus syriacus. Utilizing UV, IR, NMR, and MS spectroscopic analyses, these chemical structures were revealed. Compounds 1 and 2 were found to posses significant anti-aging properties on the human neutrophil elastase (HNE) assay, exhibiting HNE inhibitory activities with $IC_{50}$ values of 5.2 and 4.6 ${\mu}M$, respectively.

Valistatin (3-Amino-2-Hydroxy-4-Phenylbutanoyl-Valyl-Valine), a New Aminopeptidase M Inhibitor, Produced by Streptomyces sp. SL20209

  • Kho, Ying-Hee;Ko, Hack-Ryong;Chun, Hyo-Kon;Jung, Myung-Chul
    • Journal of Microbiology and Biotechnology
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    • v.5 no.1
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    • pp.36-40
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    • 1995
  • Valistatin, a new inhibitor of aminopeptidase M(AP-M) was discovered in the culture broth of Streptomyces sp. SL20209 isolated from a soil sample. The inhibitor was purified by extraction with n-butanol and the various column chromatographies, and then isolated as whitish powder. The $^1 H-and ^1 H, ^1 H-COSY$ NMR studies, amino acid analysis, and fragmentation patterns by FAB-MS suggested the presence of one 3-amino-2-hydroxy-4-phenylbutanoic acid and two valine residues in the inhibitor. Thus, the structure of valistatin was determined as 3-amino-2-hydroxy-4-phenylbutanoyl-valyl-valine. Valistatin has the molecular formular $C_20H_31N_3 O_5$ (MW 394), and its $IC_50$ value against hog kidney AP-M was determined to be 3.12 $mu g/ml$.

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Neuraminidase Inhibitors from Mushroom Microphorus affinis

  • Kim, Kyung-Bum;Kim, Sang-In;Song, Kyung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.778-782
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    • 2003
  • In the course of screening anti-influenza agents from natural products, four neuraminidase inhibitors were isolated from the methanol extract of mushroom Microphorus affinis by purification using solvent partition, silica gel column chromatography, Sephadex LH-20, and semi-preparative HPLC. The chemical structures of these compounds were identified as ${\alpha}-lupeol$, methyl linoleate, methyl palmitate, and methyl oleate by means of spectral data including GC-MS, $^1H-,\;and\;^{13}C-NMR\;with\;IC_{50}$ values of 5.65, 7.07, 7.12, and $7.52\;\mu\textrm{M}$, respectively. They did not inhibit other glycosidases such as glucosidase, mannosidase, and galactosidase, indicating that they were relatively specific inhibitors of neuraminidase. The relationship between the fatty acid structure and inhibitory activity was investigated. The result showed that, in the case of an aliphatic linear hydrocarbon skeleton, at least one carboxyl (presumably any carbonyl) moiety and sixteen carbons were the necessary requirements for potent inhibition, whereas saturated, unsaturated, free, and ester forms did not have any significant effect on the activity.

An Antitumor Component from Fomitiporia ellipsoidea

  • Zan, Lifeng;Bao, Haiying;Bau, Tolgor;Liu, Hanbin;Cui, Baokai
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1482-1485
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    • 2012
  • A natural furan derivative was isolated from the methanolic extract of the fruit bodies of Fomitiporia ellipsoidea. Its chemical structure was elucidated as methyl 3,5-dioxo-1,3,5,7-tetrahydrobenzo[1,2-c:4,5-c']difuran-4-carboxylate by means of extensive NMR and MS data analysis, and named as fomitiporiaester A (1). Compound 1 showed significant antitumor activity to hepatoma $H_{22}$ in vivo, and the inhibition rates were 42.94%, 49.17%, and 58.15% at concentrations of 5, 10, and 20 mg/kg, respectively. Compound 1 showed weak cytotoxic activities against the human hepatoblastoma (HepG-2) and human oophoroma (Skov 3) cell lines with$IC_{50}$ values of more than $100{\mu}M$.

Flavonoids from Thyrsanthera suborbicularis and Their NO Inhibitory Activity

  • Song, Hyuk-Hwan;Khiev, Piseth;Chai, Hee-Sung;Lee, Hyeong-Kyu;Oh, Sei-Ryang;Choi, Young Hee;Chin, Young-Won
    • Natural Product Sciences
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    • v.18 no.4
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    • pp.273-278
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    • 2012
  • Further phytochemical investigation on the whole plant of Thyrsanthera suborbicularis, collected in Cambodia, led to kaempferol (1), vitexin (2), apigenin-7-O-neohesperidoside (3), chrysoeriol-7-O-${\beta}$-D-glucopyranoside (4), isorhamnetin 3-O-rutinoside (5), kaempferol-3-O-[${\alpha}$-L-rhamnopyranosyl-(13)-${\alpha}$-L-rhamnopyranosyl-(16)-${\beta}$-D-galactopyranoside (6), kaempferol-3-O-${\alpha}$-L-rhamnopyranosyl(12)-O-[${\alpha}$-L-rhamnopyranosyl (16)]-${\beta}$-D-glucopyranoside (7), kaempferol-3-O-[6"-O-(E)-p-coumaroyl]-${\beta}$-D-glucopyranoside (8), kaempferol-3-O-[6"-O-(E)-p-coumaroyl]-${\beta}$-D-galactopyranoside (9), and amentoflavone (10). All the structures were confirmed by the interpretation of NMR (1D and 2D) and MS data, and comparison with the published values. Of the isolated compounds 1 - 10, compounds 8 and 10 displayed the inhibitory activity against NO production in LPS-induced Raw 264.7 cells with $IC_{50}$ values, 3.56 and $15.73{\mu}M$, respectively.

Cancer Cell Growth Inhibition of Lanostane-type Triterpenoids Isolated from Ganoderma gibbosum (칠황버섯으로부터 분리한 Lanostane-type Triterpenoid의 암세포성장 억제효과)

  • Kim, Donghwa;Lee, Sang Kook;Park, Hee-Juhn
    • Korean Journal of Pharmacognosy
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    • v.51 no.1
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    • pp.36-40
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    • 2020
  • The CHCl3 fraction of the MeOH extract of Ganoderma gibbosum (Ganodermataceae) exhibited cytotoxic activity on five cancer cell lines (MDA-MB-231, SK-hep1, A549, HCT116, and SNU638). Six highly oxygenated lanostane-type triterpenoids (lanostanoids) were isolated by column chromatography to test cytotoxicity on cancer cells. The five known lanostanoids were identified as gibbosic acids A, B, D, G, and H by comparison of molecular ion peaks with the literature data. The structure of a new lanostanoid, gibbosic acids I, was identified to be 3β,8β,15β,20S-tetrahydroxy-7,12,23-trioxolanost-9(11)-en-26-oic acid on the basis of NMR and MS spectroscopy. The three lanostanoids of gibbosic acids A, H, and I of the six isolates significantly suppressed the growth of cancer cells. In particular, the IC50 of gibbosic acid H was prominently low ranging from 2.64-6.56 μM.

Purification and Characterization of Lipase Inhibitor from Desmodium oxyphyllum DC. (도둑놈의 갈고리의 지질분해효소 저해물질의 정제 및 특성)

  • Lee, Jong-Kug;Kang, Min-Gu;Baek, Seung-Ye;Ahn, Young-Geun;Lee, Jong-Soo
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.291-294
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    • 2011
  • A lipase inhibitor from Desmodium oxyphyllum DC. was purified by methanol extraction, systematic solvent extraction, silica gel column chromatography, $C_{18}$ solid phase extraction chromatography and RP-HPLC. We obtained the purified lipase inhibitor with 182 ng($IC_{50}$) of lipase inhibitory activity for a 0.06% yield. Its molecular weight was estimated to be 655.37 Da from an instrumental analysis of MALDI-TOF-MS and it was identified copper-3,5-dibromo-2-hydroxybenzoic acid ($C_{14}H_8Br_4CuO_6$) by $^1H$, $^{13}C$ NMR analysis.