트리테르페노이드의 코르티코이드 유사활성물질에 관한 연구(II) 11-옥소올레아놀산 및 유도체의 합성과 그 항염증 활성

한병훈,조명선,한용남,박명환,Han, Byung-Hoon,KimCho, Myung-Sun,Han, Yong-Nam,Park, Myung-Hwan 생화학분자생물학회 1984 한국생화학회지 Vol.17 No.1

11-oxo-olenolic acid [I] 및 유도체를 합성하고 이 화합물들의 corticoid-$5{\beta}$-reductase 저해활성과 항염증 활성을 연구하였다. [I]을 $CH_2N_2$로 메칠화하여 11-oxo-oleanolic acid methylester [II] $C_{31}H_{48}O_4$, mp $196{\sim}198^{\circ}$를 얻었고, oleanolic acid를 $CrO_3$-pyridine으로 산화하여 3-oxo-oleanolic acid [III] $C_{30}H_{46}O_3$, $198{\sim}202^{\circ}$를 얻고 [III]을 다시 $CrO_3$-초산으로 산화하여 3, 11-dioxo-olenolic acid [IV] $C_{30}H_{44}O_4$, mp $268{\sim}270^{\circ}$, UV (${\lambda}max$) 252nm와 부산물로 3, 11-dioxo-oleanolic acid lactone [V] $C_{30}H_{44}O_4$, mp $265{\sim}267^{\circ}$를 얻었다. [IV]는 [I]에 비하여 corticoid-$5{\beta}$-reductase를 강력히 저해하며 $K_i$값은 [IV]가 $4.2{\times}10^{-4}M$, [I]이 $4.6{\times}10^{-4}M$로 측정되었다. 이 화합물의 항염증 작용을 흰쥐의 카라게닌 유발부종시험법으로 측정한 결과 30mg/kg.s.c. 용량에서 hydrocortisone이 66% 부종을 억제한데 비하여 [I]은 63%, [II]는 78% [IV]는 70%, [V]는 66% 억제하였다. 이상의 연구결과로부터 구조와 활성의 관계를 고찰할 때 올레아놀산의 탄소 11 및 12 위치의 $\alpha,\beta$-unsaturated ketone 구조가 코르티코이드 유사활성에 필수적이며 탄소3위치의 수산기 보다 케톤기가 더 큰활성을 나타나게 하고 C 및 D 환의 락톤 형성에 의한 구조변화는 이 활성증가를 감소시키고 있다. 가장 강한 활성을 나타낸 [II]의 경우는 소수성의 증가에 기인된 것으로 추정된다. The effect of 11-oxo-oleanlic acid [I] and its derivatives on carrageenin-induced edema in rat kind paw was investigated. The following derivatives were synthesized ; 11-oxo-oleanolic acid methylester [II] $C_{31}H_{48}O_4$ mp $196-198^{\circ}$ was derived from [I] by methylation with $CH_2N_2$. 3-Oxo-oleanolic acid [III] $C_{30}H_{46}O_3$, mP $198-202^{\circ}$ was obtained from oleanolic acid by $CrO_3$-pyridine oxidation. Oxidation of [III] by $CrO_3$-HAc produced 3, 11-dioxooleanolic acid [IV] $C_{30}H_{44}O_4$, mp $268-270^{\circ}$ VU ($\lambda$ max) 252 nm and 3, 11-dioxo-oleanolic acid lactone [V] $C_{30}H_{44}O_4$, mp $265-267^{\circ}$ as a by-product. [IV] showed strong competitive inhibition against corticoid-$5{\beta}$-reductase, $K_i=4.2{\times}10^{-4}M$, compared to $4.6{\times}10^{-4}M$ of [I]. The anti-inflammatory activities of 11-oxo-oleanolic acid derivatives were evaluated by rat kind paw carrageenin edema test after s.c, injection with 30mg/kg doses. [I], [II], [IV] and [V] suppressed significantly the edema volumes by 63, 78, 70 and 66%, respectively, compared to 66% of hydrocortisone. The above results suggest that the replacement of OH group in $C_3$ position by ketone group increased the corticomimetic activity, but that the change on the steric structure in C and D rings by lactone formation reduced the activity increase. [II] showed the most powerful activities probably due to increased lipophilicity.

미성숙 감귤 과피 초임계 추출물의 성분 분석과 자궁암세포 성장억제효능

문정용(Moon, Jeong Yong),송연우(Song, YeonWoo),현호봉(Hyun, Ho Bong),김소미(KimCho, Somi) 한국산학기술학회 2015 한국산학기술학회논문지 Vol.16 No.12

본 연구의 목적은 제주도 재래감귤인 팔삭과 이예감 초임계 추출물에 대한 인간 자궁암 세포인 HeLa 세포에서의 성장억제 효능을 탐색하고, 각 추출물에서의 유효 활성 성분을 분석하는데 있다. HeLa 세포에서의 성장억제 및 세포사멸 효능을 탐색하기 위해 MTT assay와 Hoechst 33342 염색을 수행하였으며, 성분분석은 가스크로마토그래피 질량분석기 (GC/MS)를 이용하였다. 두 종류의 미성숙 감귤 과육, 과피 초임계 추출물에 대한 HeLa 세포의 성장 억제 효능을 비교해 본 결과, 과피 추출물은 팔삭과 이예감 모두 비슷한 세포사멸 효능을 나타냈다. 과피 추출물을 처리한 세포에서는 농도 의존 적으로 성장률이 억제되었으며, 처리 농도 400 μg/mL에서 팔삭과 이예감 추출물이 각각 87.16%와 92.95%의 세포사멸 효능 을 나타냈으나, 과육 추출물의 세포 성장 억제 효능은 처리 농도 400 μg/mL까지 관찰되지 않았다. Hoechst 33342 염색을 통해 apoptotic body 형성을 현미경으로 관찰한 결과, 100, 200 μg/mL 과육 초임계 추출물을 처리한 세포에서는 apoptpotic body를 관찰할 수 없었으나, 동일한 농도의 과피 추출물을 처리한 세포의 경우에서는 apoptotic body가 현저하게 증가하는 것을 확인 할 수 있었다. GC/MS 분석을 통해 미성숙 팔삭 과육과 과피 초임계 추출물에서 각각 27개와 31개의 화합물을 검출하였으며, 미성숙 이예감 과육과 과피 초임계 추출물에서는 각각 27개와 29개의 화합물을 검출하였다. 팔삭 과피 초임계 추출물에는 1,1,4,4-Tetramethyl- 2-tetralone(20.86%), alloimperatorin(8.15%), limonene (11.23%), auraptene(7.29%) 등이 주로 함유되어 있었으며, 이예감 과피 초임계 추출물에는 limonene(22.19%), linalool(11.23%), γ-sitosterol(9.12%) 등이 주로 함유되 어 있었다. This study was performed to investigate the antiproliferative activities of supercritical extracts from phalsak(Citrus hassaku Hort ex Tanaka) and yeagam(Citrus iyo Hort. ex Tanaka) against human cervical carcinoma HeLa cells and the chemical compositions of the extracts. The anticancer properties of supercritical extracts were demonstrated using the MTT assay and Hoechst 33342 staining and the compositional analyses were conducted by using gas chromatography-mass spectrometry(GC-MS). The peel extracts of both species exhibited similar antiproliferative effect. The antiproliferative activity of the flesh extracts was not detected up to 400 μg/mL, whereas peel extracts of phalsak and yeagam reduced cell viability with 87.16% and 92.95% at 400 μg/mL, respectively. There was a dramatic increase of the apoptotic body formation in the cell treated with peel extracts while no apoptotic body formation detected in the cell treated with flesh extracts at 100, 200 μg/mL. By GC-MS analysis, 27 and 31 kinds of compounds identified in flesh and peel of phalsak, while 27 and 29 kinds of compounds were identified in flesh and peel of yeagam, respectively. 1,1,4,4-Tetramethyl- 2-tetralone(20.86%), alloimperatorin(8.15%), limonene(11.23%), and auraptene(7.29%) were major in peel of phalsak, whereas limonene(22.19%), linalool(11.23%), and γ -sitosterol(9.12%) were major in peel of yeagam. som

Characterization of Volatile Aroma-active Compounds in Dangyooja (Citrus grandis Osbeck)

Chung, Hyun,Chung, Won-Young,Yoo, Eun-Sook,KimCho, So-Mi,Oh, Sea-Kwan,Kim, Young-Suk The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.1

Volatile components in peel and flesh of Dangyooja (Citrus grandis Osbeck) were isolated by solvent-assisted flavor evaporation (SAFE) followed by analysis using gas chromatography-mass spectrophotometry (GC-MS). Twenty-four components, including 4 alcohols, 6 aldehydes, 14 terpene hydrocarbons, and 16 components, including 1 alcohol, 1 aldehyde, 8 terpene hydrocarbons, 4 aromatic compounds, and 2 hydrocarbons were found in peel and flesh of Dangyooja, respectively. Limonene and myrcene were dominant in both parts of Dangyooja, although more volatile components were detected in peel of Dangyooja. Aroma-active compounds in peel and flesh of Dangyooja were also evaluated by aroma extract dilution analysis. Geranial (pineapple-like) showed the highest flavor dilution factor, followed by citronellal (herbaceous) and ${\alpha}$-pinene (pine tree-like). In contrast, ${\alpha}$-pinene (pine tree-like) and myrcene (floral) were the major aroma-active components in flesh of Dangyooja.

Biological Effects of Various Solvent Fractions Derived from Jeju Island Red Sea Cucumber (Stichopus japonicus)

Park, Soo-Yeong,Lim, Hee-Kyoung,Lee, Seog-Jae,Kimcho, So-Mi,Park, Sang-Gyu,Cho, Moon-Jae The Korean Society for Applied Biological Chemistr 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.5

Red variants of S. japonicus show unique ecological characteristics and are indigenous to Jeju Island in South Korea. Various biological activities of red sea cucumber extracts (RSCEs) were evaluated. In comparison with positive controls, anti-oxidant activity of RSCEs was very low. In HL-60 and HT-29 cells, chloroform and ethyl acetate (EtOAc) fractions showed higher than 80 and 60% growth inhibition, respectively. Nuclear condensation and increased pro-apoptotic signaling revealed that RSCEs triggered apoptosis. EtOAc fractions also showed strong anti-inflammatory effects at sub-lethal concentrations in lipolysaccharide-stimulated RAW264.7 cells and suppressed more than 90% of nitric oxide (NO) and prostaglandin 2 productions at 50 ${\mu}g$/mL by inhibiting inducible NO synthase and cydooxygenase-2. Water-soluble fractions showed good antibacterial effects against Staphylococcus aureus and Staphylococcus epidermidis.

Induction of Apoptosis in SNU-16 Human Gastric Cancer Cells by the Chloroform Fraction of an Extract of Dangyuja (Citrus grandis) Leaves

Moon, Jeong-Yong,Kim, Ha-Na,Cho, Moon-Jae,Chang, Weon-Young,Kim, Cheong-Tae,KimCho, So-Mi The Korean Society for Applied Biological Chemistr 2009 Applied Biological Chemistry (Appl Biol Chem) Vol.52 No.2

The anticancer effect of Dangyuja (Citrus grandis Osbeck) leaf extract was investigated using SNU-16 human gastric cancer cells. Maximum cytotoxicity was observed from the chloroform fraction (CF) of the extract. Cell death was dose-dependent ($IC_{50}$=ca. 92.15 ${\mu}g/mL$), and was characterized by apoptotic body formation and DNA fragmentation. Flowcytometric analysis showed that treatment of CF resulted in a marked accumulation of cells in the sub-G1 phase. The induction of apoptosis was confIrmed by caspase-3 activity assays and immunoblotting using antibodies against Bid, Bcl-2, Bax, poly (ADP-ribose) polymerase, caspase-9, caspase-8, caspase-7, and caspase-3. Analyses of the CF by gas chromatography (GC) and GC-mass spectrometry tentatively identifIed 21 compounds, including linoleic acid, linolenic acid, palmitic acid, ${\alpha}$-amyrin, ${\gamma}$-sitosterol, dihydrolanosterol, methyl palmitate, and nobiletin. These results provided the fIrst evidence that the CF of a Dangyuja leaf extract induces apoptosis in SNU-16 cells. Our findings may lead to new strategies for the treatment of human gastric cancer.

Mango (Mangifera indica L.) Peel Extracts Inhibit Proliferation of HeLa Human Cervical Carcinoma Cell via Induction of Apoptosis

Ali, Md. Ramjan,Yong, Moon-Jeong,Gyawali, Rajendra,Mosaddik, Ashik,Ryu, Youn-Chul,KimCho, So-Mi The Korean Society for Applied Biological Chemisty 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.3

Flesh and peel extracts of five mango varieties were compared for their antioxidant and antiproliferative activities. Fozli peel (FP) extract was found to have the highest antioxidant activity and to significantly inhibit the proliferation of HeLa human cervical carcinoma cells in a dose-dependent manner. Hoechst 33342 staining revealed apoptotic nuclei with condensed chromatin, and cell growth arrest was confirmed by flow cytometry. Apoptotic signaling induced by FP extract was characterized by an increased Bax/Bcl-2 ratio and down-regulation of Bcl-2. Moreover, FP extract treatment triggered the proteolytic activation of caspases-3, -8, and -9 and the degradation of poly ADP-ribose polymerase in HeLa cells. These results indicate that FP is an excellent source of phenolic and/or flavonoid compounds, and may have applications in the treatment of human cervical carcinoma and as an antioxidant in the food and pharmaceutical industries. Gas chromatography-mass spectrometry analysis of FP extract revealed that 2,5-dihydroxyphenol was detected with highest amount and other compounds such as 5-hydroxymethyl-2-furancarboxaldehyde, pentadecanoic acid, (Z,Z)-9,12-octadecadienoic acid, and 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one were also found in high amounts.

Anti-Cancer Effect of Ginsenoside F₂ against Glioblastoma Multiforme in Xenograft Model in SD Rats

Shin, Ji-Yon,Lee, Jung-Min,Shin, Heon-Sub,Park, Sang-Yong,Yang, Jung-Eun,KimCho, So-Mi,Yi, Tae-Hoo The Korean Society of Ginseng 2012 Journal of Ginseng Research Vol.36 No.1

The glioblastoma multiforme (GBM) is the most common malignant brain tumor in adults. Despite combination treatments of radiation and chemotherapy, the survival periods are very short. Therefore, this study was conducted to assess the potential of ginsenoside $F_2$ (F2) to treat GBM. In in vitro experiments with glioblastoma cells U373MG, F2 showed the cytotoxic effect with $IC_{50}$ of 50 ${\mu}g/mL$ through apoptosis, confirmed by DNA condensation and fragmentation. The cell population of cell cycle sub-G1 as indicative of apoptosis was also increased. In xenograft model in SD rats, F2 at dosage of 35 mg/kg weight was intravenously injected every two days. This reduced the tumor growth in magnetic resonance imaging images. The immunohistochemistry revealed that the anticancer activity might be mediated through inhibition of proliferation judged by Ki67 and apoptosis induced by activation of caspase-3 and -8. And the lowered expression of CD31 showed the reduction in blood vessel densities. The expression of matrix metalloproteinase-9 for invasion of cancer was also inhibited. The cell populations with cancer stem cell markers of CD133 and nestin were reduced. The results of this study suggested that F2 could be a new potential chemotherapeutic drug for GBM treatment by inhibiting the growth and invasion of cancer.