• Journal of Microbiology and Biotechnology
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• 제18권10호
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• pp.1659-1662
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• 2008

Pseudomonas chlororaphis O6 exhibits induced systemic resistance (ISR) against P. syringae pv. tabaci in tobacco. To identify one of the ISR metabolites, O6 cultures were extracted with organic solvents, and the organic extracts were subjected to column chromatography followed by spectroscopy analyses. The ISR bioassay-guided fractionation was carried out for isolation of the metabolite. High-resolution mass spectrometric analysis of the metabolite found $C_{9}H_{9}O_{3}N$ with an exact mass of 179.0582. LC/MS analysis in positive mode showed an $(M+H)^{+}$ peak at m/z 180. Nuclear magnetic resonance ($^{1}H,\;^{13}C$) analyses identified all protons and carbons of the metabolite. Based on the spectroscopy data, the metabolite was identified as 4-(aminocarbonyl) phenylacetate (4-ACPA). 4-ACPA applied at 68.0 mM exhibited ISR activity at a level similar to 1.0 mM salicylic acid. This is the first report to identify an ISR metabolite produced by P. chlororaphis O6 against the wildfire pathogen P. syringae pv. tabaci in tobacco.

• Bulletin of the Korean Chemical Society
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• 제31권7호
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• pp.1931-1936
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• 2010

Discovery and isolation of compounds capable of blocking the interactions between VCAM-1 and VLA-4, a major pair of adhesion molecules contributing to the different steps of leukocyte migration across the endothelium in inflammatory responses, has been a major goal of this lab. Through bioactivity-guided fractionation, five saikosaponins were subsequently isolated from the methanol extracts of the roots of Bupleurum falcatum L. Their structures were elucidated by spectroscopic analysis ($^1H-$, $^{13}C$-NMR and 2D-NMR), as follows, saikosaponins: A (1); D (2); C (3); B3 (4); B4 (5). Compounds 1 and 2 inhibited interaction of sVCAM-1 and VLA-4 of THP-1 cells with respective $IC_{50}$ values of 7.8 and 1.7 ${\mu}M$. The aglycone structure of 2 also showed cell adhesion inhibitory activity with an $IC_{50}$ value of 21.1 ${\mu}M$. With these results, we suspect these two saikosaponins from the Bupleurum falcatum L. roots to be prime candidates for therapeutic strategies towards inflammation.

• 생명과학회지
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• 제16권2호
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• pp.282-288
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• 2006

새송이버섯으로부터 활성추적법으로 항암활성이 있는 물질인 에르고스테롤 프록사이드를 분리하였다. 이 스테롤의 구조는 분광법과 NMR법으로 확인하였으며 분자식은 $C_{28}H_{44}O_3$이었다. 폐암과 난소암에 $IC_{50}$값은 각각 $7{\mu}M$과 $14{\mu}M$이었다. DNA단편화 실험에서 이 화합물은 암세포의 chromosimal DNA 를 사닥다리모양으로 분해하였고, 세포 분열주기의 억제실험에서 G1단계를 억제함을 관찰하였다.

• BMB Reports
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• 제43권4호
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• pp.268-272
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• 2010

Aldo-keto reductase family 1 B10 (AKR1B10) is a member of the NADPH-dependent aldo-keto reductase (AKR) superfamily, and has been considered to be a potential cancer therapeutic target. Total extract from the bark of Rhus verniciflua (Toxicodendron vernicifluum (Stokes)) showed AKR1B10 inhibitory activity. To identify the active compounds from R. verniciflua responsible for AKR1B10 inhibition, nine compounds were isolated via bioactivity-guided isolation and tested for their effects against recombinant human AKR1B10 (rhAKR1B10). Results showed that butein, isolated from the ethyl acetate fraction, was most able to inhibit rhAKR1B10. The inhibitory rate of butein against rhAKR1B10 was 42.86% at $1\;{\mu}M$ with an IC50 value of $1.47\;{\mu}M$, and enzyme kinetic analysis revealed its inhibition mode to be uncompetitive.

• 생약학회지
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• 제39권4호
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• pp.347-351
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• 2008

The methanol (MeOH) extract of the rhizome of Alpinia officinarum Hance (Zingiberaceae) demonstrated a potent inhibition on the proliferation of cultured human tumor cell lines such as MES-SA (human uterine carcinoma cell line), MESSA/DX5 (multidrug resistant subline of MES-SA), HCT-15 (human colorectal adenocarcinoma cell line), HCT15/CL02 (multidrug resistant subline of HCT15). The MeOH extract was fractionated into four portions by serial solvent partition, ie., methylene chloride (CH2Cl2) soluble part, ethylacetate (EtOAc) soluble part, n-butanol (BuOH) soluble part and remaining water layer. Among them, the $CH_2Cl_2$ soluble part of the extract exhibited a most potent inhibition on the proliferation of tested tumor cell lines. Bioassay-guided fractionation of the $CH_2Cl_2$ soluble part led to the isolation of five diarylheptanoid and two flavonoid constituents, i. e., galangin (1), 7-(4"-hydroxy-3"-methoxyphenyl)-1-phenylhept-4-en-3-one (2), 1,7-diphenyl-5-hydroxy-3-heptanone (3), trans,trans-1-(3'-methoxy-4'-hydroxyphenyl)-7-phenyl-5-ol-4,6-dien-3-heptanone (4), 5-methoxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (5), kaempferide (6), 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (7). Structures of the isolated active components (1 - 7) were established by chemical and spectroscopic means.

• Archives of Pharmacal Research
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• 제28권1호
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• pp.22-27
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• 2005

Bioassay-guided fractionation of methanol extract of Reynoutria sachalinensis flower using DPPH assay has led to the isolation of three anthraquinones and three flavonoids. Their structures were identified as emodin (1), emodin-8-O-$\beta$-D-glucopyranoside (2), physcion-8-O-$\beta$-D­glucopyranoside (3), quercetin-3-O-$\alpha$-L-arabinofuranoside (4), quercetin-3-O-$\beta$-D-galactopyra­noside (5), and quercetin-3-O-$\beta$-D-glucuronopyranoside (6) by comparing their physicochemical and spectral data with those published in literatures. All isolated compounds were evaluated for antioxidant activities with free radical 1, 1-diphenyl-2-picrylhydrazyl (DPPH) scavenging, superoxide radical scavenging and $Cu^{2+}$-mediated low density lipoprotein (LDL) oxidation assay. The results demonstrated that three flavonoids, 4, 5, and 6 had remarkable antioxidant activities with the $IC_{50}$ values of 64.3, 54.7, and 46.2${\mu}M$ (DPPH scavenging), the $IC_{50}$ values of 6.0, 6.7, and $4.4{\mu}M$ (superoxide radical scavenging) and the $IC_{50}$ values of 3.8, 3.2, and 5.4${\mu}M$ against LDL oxidation, respectively.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.161-166
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• 2013

At present, acetylcholinesterase (AChE) inhibitors are the first group of drugs to treat mild to moderate Alzheimer's disease (AD). Although beneficial in improving cognitive and behavioral symptoms, the effectiveness of AChE inhibitors has been questioned since they do not delay or prevent neurodegeneration in AD patients. Therefore, in the present study, in order to develop new and effective anti-AD agents from lichen products, both the AChE inhibitory and the neuroprotective effects were evaluated. The AChE inhibitory assay was performed based on Ellman's reaction, and the neuroprotective effect was evaluated by using the MTT method on injured PC12 cells. One AChE inhibitor ($IC_{50}$ = 27.1 ${\mu}g/ml$) was isolated by means of bioactivity-guided isolation from the extract of lichen-forming fungus Cladonia macilenta, which showed the most potent AChE inhibitory activity in previous screening experiment. It was then identified as biruloquinone by MS, and $^1H$- and $^{13}C$-NMR analyses. The inhibitory kinetic assay suggested that biruloquinone is a mixed-II inhibitor on AChE. Meanwhile, biruloquinone improved the viability of the $H_2O_2$- and ${\beta}$-amyloid-injured PC12 cells at 1 to 25 ${\mu}g/ml$. The protective effects are proposed to be related to the potent antioxidant activities of biruloquinone. These results imply that biruloquinone has the potential to be developed as a multifunctional anti- AD agent.

• Advances in Traditional Medicine
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• 제10권3호
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• pp.200-207
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• 2010

The fruits of Piper cubeba L. are used traditionally to treat respiratory disorders in Indonesia. In order to determine the compounds responsible for this activity, the fruits were extracted with nhexane followed by ethanol to give n-hexane and ethanol extracts. Based on tracheospasmolytic assay on these two extracts, the n-hexane extract was more active to inhibit trachea contraction than that of ethanol extract. Upon bioassay guided isolation of the n-hexane extract, a tracheospasmolytic active compound was isolated and identified as dihydrocubebin [(3,4),(3',4')-bis-methylenedioxy-9,9'dihydroxylignan] $(\underline{1})$. Compound $\underline{1}$ was tested further for its ability to inhibit histamine released from mast cells, using rat basophilic leukemia (RBL-2H3) cell line and rat peritoneal mast cells RPMCs) as models; and $DNP_{24}$-BSA, thapsigargin, ionomycin, compound 48/80 and PMA were used as inducers for histamine released from mast cell. The test result showed that $\underline{1}$ inhibited histamine release from RBL-2H3 cells induced by $DNP_{24}$-BSA, thapsigargin and ionomycin. In addition, $\underline{1}$ suppressed histamine release from RPMC induced by either thapsigargin or ionomycin. However, $\underline{1}$ did not inhibit histamine release from RPMC induced by either compound 48/80 or combination PMA-sub optimum dose of ionomycin. Therefore, it was concluded that the inhibitory effects of $\underline{1}$ on the histamine released from mast cells may involve mechanisms related to intracellular $Ca^{2+}$ signaling events or downstream processes of intracellular $Ca^{2+}$ signaling in mast cells.

• 생약학회지
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• 제36권3호통권142호
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• pp.201-204
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• 2005

The rhizome extract of Atractylodes japonica Koidzumi(Compositae) exhibited a particular inhibition on the proliferation of cultured human tumor cell lines, in vitro. Thus, the intensive phytichemical investigation of the MeOH extract of Atractylodes japonica have been conducted by the way of activity-guided purification. The repeated column chromatographic separation of the n-hexane soluble part of extract resulted in the isolation of four sesquiterpenes (1-4) and a polyacetylene component (5). Chemical structures of them were identified as atractylon (1), atractylenolide Ⅰ(2), atractylenolide Ⅲ(3), eudesma-4(15),7(11)-dien-8-one (4) and 1,3-diacetyl-atractylodiol (5) by spectroscopic means. Among the isolates, compound 2-4 were shown to give moderate inhibitory effect in a dose dependent manner on the proliferation of cultured human tumor cell lines such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCT 15(colon), respectively.

• 한국식품영양과학회지
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• 제37권8호
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• pp.979-984
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• 2008

본 연구에서는 머위 엽병의 에탄올 추출물을 용매 극성에 따라 hexane, chloroform, ethyl acetate 및 n-butanol로 분배한 분획물에 대해 전자공여능, 지질과산화 억제능 및 lipoxygenase 저해효과 효과를 살펴봄으로써 항산화능을 측정하였다. 분획물 중 ethyl acetate 분획물의 경우 $100\;{\mu}g/mL$ 농도에서 전자공여능, 지질과산화 억제능 및 lipoxygenase 저해능에서 각각 65.41%, 37.04%, 50.67%로 가장 우수한 항산화능을 나타내었다. 이를 근거로 하여 ethyl acetate 분획물로부터 머위 엽병의 항산화 활성 성분을 규명하기 위하여 open column chromatography를 이용하여 화합물 PJ-4를 분리, 정제하였으며, 1H- 및 13C-NMR 분석을 통해 kaempferol로 동정하였다. 단리된 kaempferol의 항산화 활성을 확인하기 위하여 앞에서 언급한 세 가지 항산화 측정법을 통해 실험한 결과, 농도 의존적으로 활성이 증가하는 양상을 보였다. 특히 $100\;{\mu}g/mL$의 농도에서 전자공여능, TBARS에 의한 지질과산화 억제 및 lipoxygenase 저해실험에서 각각 65.76%, 43.47%, 58.60%로 높은 항산화 활성을 나타내었다. 이와 같은 결과를 통해 kaempferol은 머위 엽병의 항산화 활성에 있어서 기여하는 물질일 것으로 보이며, 이로써 머위엽병을 이용한 천연항산화제 개발 및 활용에 긍정적인 가능성을 제시할 것으로 사료된다.