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標題: | 第一部分 山欖葉部之成分研究(III):分離、三萜皂苷之分析與其醣基之確認
第二部分 大葉山欖果仁之活性成分研究 Part I Chemical investigation of Planchonella obovata leaf (III): isolation, analysis of triterpenoid glycosides and confirmation of their glycon moieties Part II Bioactive constituents from Palaquium formosanum kernel |
作者: | Hsin-Yi Chen 陳欣苢 |
指導教授: | 李水盛 |
關鍵字: | 山欖,大葉山欖,山欖科,三?皂?,1D TOCSY,細胞毒性,原花青素,旋轉異構物,氫暨碳譜標定, Planchonella obovata,Palaquium formosanum,Sapotaceae,triterpenoid glycosides,1D TOCSY,cytotoxicity,proanthocyanidins,rotamers,1H & 13C NMR assignment, |
出版年 : | 2017 |
學位: | 博士 |
摘要: | 第一部分 山欖葉部之成分研究(III):分離、三萜皂苷之分析與其醣基之確認
山欖(Planchonella obovata)為常青樹木,是臺灣三種山欖科植物其中之一。本論文為延續本實驗室先前對其正丁醇可溶之高極性成分探討。使用Sephadex LH-20、逆向管柱層析得到2個新化合物,6β-hydroxy-conyzasaponin G (8)和glycerol 1,2-disinapate 3-α-glucuronide (9),和5個已知化合物。 通過薄層層析法分析Mi-saponin B和C酸水解產物,得知皂苷裡單醣的組成。將這些單醣做成trimethylsilyl L-cysteine衍生物,接著利用氣相層析分析,即可鑑定出這些單醣的絕對立體結構。這些單醣為D-apiose、L-arabinose、D-xylose、D-glucose和L-rhamnose。 第二部分 大葉山欖果仁之活性成分研究 山欖科植物大葉山欖(Palaquium formosanum)為高大之喬木,原生於菲律賓和台灣。其果仁酒精萃取物的正丁醇可溶層對PC-3前列腺癌細胞具有好的細胞毒性,因而將正丁醇可溶層經Sephadex LH-20分離。富含皂苷的組分具有細胞毒殺性,因而針對此一部分進行分離,得到1個新的化合物,3'-O-rhamnopyranosyl-arganin C (10),以及11個已知的oleanane-type皂苷(11–21)。這些化合物的1H和13C訊號相當複雜,需要經由1D NMR (1H和13C)、2D NMR (1H−1H COSY、NOESY、HSQC和HMBC),以及選擇性激發1D TOCSY圖譜進行繁複的分析而確立。本研究修正不少文獻之1H和13C NMR標定,而且這些圖譜是在methanol-d4紀錄,方便日後用於鑑定類似的三萜皂苷之參考。這些化合物加上五個來自山欖的三萜皂苷一起做活性試驗,結果顯示arganin C具有好的抗PC-3前列腺癌細胞活性,IC50為13.8 μM。本研究亦歸納這些三萜皂苷結構與抗PC-3前列腺癌細胞活性關係。 另一方面,正丁醇可溶層對甲型葡萄糖水解酶亦有甚佳之抑制活性,在10 μg/mL下達87.8%抑制率。該層依活性導引進行分離,得到10個類黃酮成分(22–31)和兩個megastigmane glycosides (32和33)。其中四個為二聚原花青素(dimeric proanthocyanidins)以4α→8方式相接,由於axial rotation使之產生旋轉異構物(rotational isomers)。因此 prodelphinidin B3 (28)、catechin-(4α→8)-gallocatechin (29)、gallocatechin-(4α→8)-gallocatechin (30)和gallocatechin-(4α→8)-epigallocate chin gallate (31), 1H和13C NMR 圖譜相當複雜。藉由2D ROESY圖譜了解rotamers的特性,再經由分析2D NMR (1H−1H COSY、ROESY、HSQC和HMBC),將這些rotamers完整的1H和13C NMR資料加以標定,此亦為首次之報告。 Part I. Chemical investigation of Planchonella obovata leaf (III): isolation, analysis of triterpenoid glycosides and confirmation of their glycon moieties Planchonella obovata is an evergreen tree, one of the three Sapotaceae plants in Taiwan. This thesis is a continuation of our previous study on the chemical investigation of the polar components in the n-BuOH soluble fraction. Further separation through Sephadex LH-20 and reversed-phase C-18 columns led to the isolation of two new compounds, 6β-hydroxy-conyzasaponin G (8) and glycerol 1,2-disinapate 3-α-glucuronide (9), and five known compounds. The monosaccharide compositions in these glycosides were determined by TLC analysis of the acid hydrolysates of Mi-saponins B and C. The absolute configurations of these monosaccharides was determined by GC analysis of the corresponding trimethylsilyl L-cysteine derivative. These monosaccharides are determined as D-apiose, L-arabinose, D-xylose, D-glucose, and L-rhamnose. Part II Bioactive constituents from Palaquium formosanum kernel Palaquium formosanum (Sapotaceae) is a tall tree indigenous to Philippines and Taiwan. The n-BuOH-soluble fraction of the ethanol extact of the P. formosanum kernel being cytotoxic against PC-3 prostate cancer cell line was further fractionated by Sephadex LH-20. The fractions rich in saponins showed cytotoxicity were further separated mainly by RP-18 columns.This effort led to the isolation of one new, 3'-O-rhamnopyranosyl-arganin C (10), and 11 known oleanane-type saponins (11–21). The 1H and 13C NMR spectra of these compounds are quite complex and their assignments were established by analysis of 1D NMR (1H and 13C), 2D NMR (1H−1H COSY, NOESY, HSQC, and HMBC), and selectively excited 1D TOCSY spectra. This work revised some reported 1H and 13C NMR assignments, and these data being recorded in methanol-d4 are friendly to the user as a reference for identifying the related triterpenoid saponins. Bioassay of these isolated compounds and five from Planchonella obovata leaf indicated arganin C to be most potent against PC-3 prostate cancer cells with the IC50 value of 13.8 μM. Some structure and activity relationships were also drawn. The n-BuOH-soluble fraction also displayed potent inhibitory activity against α-glucosidase (87.8% at 10 μg/mL). Further investigation of this fraction via the bioassay-guide approach yielded ten flavonoids (22–31). Four of them (28–31) are dimeric proanthocyanidins with 4α→8 linkage, arising rotational isomers from axial rotation. Thus, the 1H and 13C NMR spectra of prodelphinidin B3 (28), catechin-(4α→8)-gallocatechin (29), gallocatechin-(4α→8)-gallocatechin (30), and gallocatechin-(4α→8)-epigallocatechin gallate (31) were quite complex. The conformations of these rotamers were identified through analysis of 2D ROESY spectra. The complete 1H and 13C NMR assignments of these rotamers were made by extensive analysis of 2D NMR (1H−1H COSY, ROESY, HSQC, and HMBC), and this is the first report for such assignments. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68531 |
DOI: | 10.6342/NTU201704053 |
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顯示於系所單位: | 藥學系 |
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