• Archives of Pharmacal Research
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• 제27권2호
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• pp.169-172
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• 2004

Activity-guided fractionation of the EtOAc-soluble extract of the stems of Couepia ulei, using a bioassay based on the induction of quinone reductase (QR) in cultured Hepa 1c1c7 mouse hepatoma cells led to the isolation of two active compounds, a new natural product, erythro-2,3-bis(4-hydroxy-3-methoxyphenyl)-3-ethoxypropan-1-o1 (1), and a known compound, evofolin-B (2), along with five inactive compounds all of known structure, viz., betulinic acid, oleanolic acid, pomolic acid, ($\pm$)-syringaresinol, and ursolic acid. These isolates were identified by analysis of physical and spectral data. Compounds 1 and 2 exhibited QR inducing activity, with observed CD (concentration required to double induction) values of 16.7 and 16.4 $\mu\textrm{M}$, respectively.

• 생약학회지
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• 제36권3호통권142호
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• pp.240-244
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• 2005

The methanol extract of the seed of Myristica fragrans (myristicaceae) demonstrated a potent inhibition on the proliferation of cultured human tumor cells such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2(melanoma), XF498 (central nerve system) and HCT-15(colon). The MeOH extract was fractionated into three portions by serial solvent partition i,e., EtOAc soluble part, BuOH soluble part and remaining water layer. Among them, the EtOAc soluble part of the extract demonstrated a potent inhibition on the proliferation of cultured human tumor cells, Bioassay-guided fractionation of the EtOAc soluble part led to the isolation of six lignan constituents, i.e., safrole(1), machilin A (2), licarin B (3), macelignan (4), mesodihydroguaiaretic acid (5) and myristargenol A (6) as well as a large amount of myristic acid as active ingredients. Structures of the isolated active components (1-6) were established by chemical and spectroscopic means.

• Journal of Microbiology and Biotechnology
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• 제30권12호
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• pp.1885-1895
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• 2020

Rumex japonicus Houtt (RJH) is a valuable plant used in traditional medicine to treat several diseases, such as scabies and jaundice. In this study, Jurkat cell growth inhibitory extracts of R. japonicus roots were subjected to bioassay-guided fractionation, resulting in the isolation of three naphthalene derivatives (3-5) along with one anthraquinone (6) and two phenolic compounds (1 and 2). Among these compounds, 2-methoxystypandrone (5) exhibited potent anti-proliferative effects on Jurkat cells. Analysis by flow cytometry confirmed that 2-methoxystypandrone (5) could significantly reduce mitochondrial membrane potential and promote increased levels of mitochondrial reactive oxygen species (ROS), suggesting a strong mitochondrial depolarization effect. Real-time quantitative polymerase chain reaction (qPCR) analysis was also performed, and the results revealed that the accumulation of ROS was caused by reduced mRNA expression levels of heme oxygenase (HO-1), catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD). In addition, 2-methoxystypandrone (5) triggered strong apoptosis that was mediated by the arrest of the G0/G1 phase of the cell cycle. Furthermore, 2-methoxystypandrone (5) downregulated p-IκB-α, p-NF-κB p65, Bcl2, and Bcl-xl and upregulated BAX proteins. Taken together, these findings revealed that 2-methoxystypandrone (5) isolated from RJH could potentially serve as an early lead compound for leukemia treatment involving intracellular signaling by increasing mitochondrial ROS and exerting anti-proliferative effects.

• 한국식품저장유통학회지
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• 제16권3호
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• pp.435-441
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• 2009

비파 씨를 MeOH로 침지 추출하여 얻어진 추출물을 n-hexane, EtOAc, n-BuOH 및 $H_2O$로 용매 분획하였다. 이중 tyrosinase, DPPH 및 SOD 활성이 상대적으로 높은 EtOAc 분획에 대해 column chromatography를 이용하여 6개의 페놀성 화합물을 분리하였다. 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 benzaldehyde (1), chlorogenic acid (2), caffeic acid (3), benzoic acid (4), ferulic acid (5) 및 amygdalin (6)으로 동정하였다. 이들 화합물중 benzaldehyde (1)는 125 mg/mL농도에서 tyrosinase저해활성이 70.8%로 나타나 positive control인 kojic acid의 56.3%보다 높게 나타났다. 또한 분리한 화합물 중 DPPH 라디칼 소거능은 12.5 mg/mL 실험농도에서 chlorogenic acid (2)와 caffeic acid (3)가 각각 43.7, 81.5%의 저해활성을 나타내었으며, 비파 씨의 SOD 활성물질은 chlorogenic acid (2)임을 확인하였다. 향후 이들 활성물질의 활성 기작에 대한 연구가 필요하며 본 연구결과는 보다 안전하고 우수한 tyrosinase 및 라디칼소거능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을뿐만 아니라 비파 씨의 식물 화학적 성분에 대한 기초자료로 이용될 수 있을 것으로 사료된다.

• 한국식품영양과학회지
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• 제44권7호
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• pp.1090-1099
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• 2015

본 연구는 잠상산물로부터 고부가가치 항당뇨 기능성 소재를 개발하기 위한 연구의 일환으로, 먼저 뽕나무 부위별 4가지(오디, 뽕잎, 상지, 상백피) 에탄올 추출물의 ${\alpha}$-glucosidase 저해활성을 측정한 결과 상백피가 가장 높은 저해활성을 나타내었으며, 그로부터 여러 column chromatography 및 NMR 기기분석을 통해 4가지 ${\alpha}$-glucosidase 저해제를 분리, 정제 및 동정한 결과는 다음과 같다. 상백피 에탄올 추출물을 Diaion HP-20, silica gel, ODS-A 및 Sephadex LH-20 column chromatography를 실시하여 4가지 화합물[Comp. 1(10 mg), Comp. 2(9.6 mg), Comp. 3(9.3 mg), 및 Comp. 4(6.5 mg)]을 분리 및 정제하였으며, 그들의 효소 저해활성을 측정한 결과 Comp. 1($IC_{50}=5.22{\mu}g/mL$), Comp. 2($IC_{50}=1.78{\mu}g/mL$), Comp. 3($IC_{50}=2.94{\mu}g/mL$) 및 Comp. 4($IC_{50}=1.54{\mu}g/mL$)로 나타났다. 그리고 그들 화합물의 동정을 위해 UV 및 NMR spectroscopy를 이용하여 구조를 분석한 결과 Comp. 1(morusin), Comp. 2(kuwanon H), Comp. 3(chalcomoracin A), Comp. 4(chalcomoracin B)로 각각 동정하였다. 이상의 연구 결과로 미루어 보아 상백피로부터 분리된 ${\alpha}$-glucosidase 저해제는 향후 당뇨 치료용 기능성 소재로 활용할 수 있을 것으로 기대된다.

• 대한한의학방제학회지
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• 제16권1호
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• pp.117-130
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• 2008

Korean traditional medicine has been used for the treatment of the various diseases based on both oriental medicinal theory and clinical trials. Thus, the prescriptions of Korean traditional medicine would be useful for the development of new therapeutics. This research focuses on the fundamental study in Korean traditional prescriptions for the development of new hepatoprotective agents. We found two prescriptions. Injinho-Tang and Osumogwa-Tang, showed the significant DPPH free radical scavenging and hepatoprotective effect, respectively. It is well-known that free radical scavenging effect is related to the prevention of various pathological events including liver injury. This paper deals with hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, free radicals scavenging on both DPPH and superoxide of above two prescriptions. Hot water extract of Injinho-Tang did not show the significant hepatoprotective effect on tacrine-induced cytotoxicity in Hep G2 cells, however, it shows the significant scavenging effects for both DPPH and superoxide radicals. On the other hand, all of the hot water extracts of constituent herbal drugs in Injinho-Tang exhibited the promising protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Of these, water extract of Rhei Rhizoma showed the most prominent effect on tacrine-induced cytotoxicity in Hep G2 cells. Bioassay-guided fractionation of Rhei Rhizoma extract has furnished four compounds, and their chemical structures have been identified by comparison of their spectral data with those of literature as chrysophanol (1), emodin (2), 3,5-dihydroxy-4'- methoxystilbene (3), and rhapontigenin (4), respectively. Among the isolated compounds, compounds 2-4 revealed the significant hepatoprotective effect in vitro when their $EC_{50}$ values compare with that of silybin, as a positive control. It also exhibited that emodin possessed the most hepatoprotective effect among these active compounds. In case of Osumogwa-Tang, its hot water extract showed the moderate protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Hot water extract of Chaenomelis Fructus, one of the constituent herbal drug of this prescription, exhibited the significant hepatoprotective effect with $EC_{50}$ value of $7.8{\pm}0.1\;{\mu}g/ml$, however, it showed strong cytotoxicity in Hep G2 cells above the concentration of $25\;{\mu}g/ml$. It was revealed that both hot water extract of Evodiae Fructus and its butanol soluble fraction showed the moderate hepatoprotective effect but concentration-dependent activity in Hep G2 assay system. Two quinolone alkaloids, evocarpine and dihydroevocarpine, also tested for their hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, however, these two compounds derived from the Evodiae Fructus as the major constituents did not show in vitro hepatoprotective effect. From these results, it would be necessary to further isolation of its hepatoprotective compounds from the butanol soluble fraction of the hot water extract of Evodiae Fructus.

• Applied Biological Chemistry
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• 제50권4호
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• pp.262-267
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• 2007

토양에는 생존하지만 현 기술로는 배양이 불가능한 미생물로부터 천연 항생제를 탐색하기 위해 한국 토양 DNA 단편들을 가진 cosmid library을 대장균에서 제작하였고 약6만개의 clone들을 대상으로 항세균 활성을 보여주는 YS92B를 최종 선발하였다. YS92B클론 배양액의 ethyl acetate추출액은 다양한 병원성 세균의 성장을 in vitro에서 강력히 저해하였다(Listeria monocytogenes, Bacillus subtilis, Pseudomonas syringae, Xanthomonas campestris pv. oryzae, Staphylococcus epidemis). 이 항균활성의 주 물질인 YS92B-VII는 ethyl acetate추출, Sephadex LH20 column chromatography와 HPLC(High Performance Liquid Chromatography)에 의해 순차적으로 분리하였으며 이 과정에서 주 활성물질은 각 피크를 항균검정으로 추적하여 최종 정제하였다. 이 물질은 NMR(Nuclear Magnetic Resornance)에 의한 구조 분석 결과에서 탄소 12개의 포화지방산인 lauric acid가 tyrosine에 결합된 N-lauroyl tyrosine임을 최종 확인하였다. 따라서 본 보고는 한국 토양에서 유래한 고유의 N-acyl amino acid synthase(NAS)유전자가 대장균에 발현되어 생산되는 N-acyl amino acid tyrosine의 특성을 밝히는 것이다.

• Archives of Pharmacal Research
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• 제26권8호
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• pp.585-590
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• 2003

Activity-guided fractionation of the EtOAc-soluble extract of the whole plants of Sida acuta using a bioassay based on the induction of quinone reductase (OR) in cultured Hepa 1c1c7 mouse hepatoma cells, led to the isolation of ten active compounds of previously known structure, quindolinone (1), cryptolepinone (2), 11-methoxyquindoline (3), N-trans-feruloyltyramine (4), vomifoliol (5), loliolide (6), 4-ketopinoresinol (7), scopoletin (8), evofolin-A (9), and evofolin-B (10), along with five inactive compounds of known structure, ferulic acid, sinapic acid, syringic acid, ($\pm$)-syringaresinol, and vanillic acid. These isolates were identified by physical and spectral data measurement. A new derivative of quindolinone, 5,10-dimethylquindolin-11-one (1a) was synthesized and characterized spectroscopically. Of the active substances, compounds 1-3 and 1a exhibited the most potent QR activity, with observed CD (concentration required to double induction) values ranging from 0.01 to 0.12 $\mu$ g/mL. Six compounds were then evaluated in a mouse mammary organ culture assay, with cryptolepinone (2), N-trans-feruloyltyramine (4), and 5,10-dimethylquindolin-11-one (1a) found to exhibit 83.3, 75.0, and 66.7% inhibition of 7,12-dimethylbenz[a]anthracene-induced preneoplastic lesions, respectively, at a dose of 10 $\mu\textrm{g}$/mL.