鐘萼木枝葉及果殼之成分研究

Abstract

本論文主要研究鐘萼木(Bretshneidera sinensis Hemsley)含有之天然物活性成分，將其枝葉及果殼部分使用酒精萃取，接著使用各種管柱色層分析方法進行分離鑑定所含之化學組成成分。實驗結果共分離了8個化合物，其中包括了5個新的含硫之生物鹼(sulfur- containing alkaloids)。 分離得到的新化合物中，共有3個屬於3-methyl-1,3-benzothiazine-2,4-dione骨架的雜環化合物(heterocyclic compounds, 1─3)、2個骨架為N-benzyl-S-methyl-thiocarbamate的含硫之生物鹼(4和5)、以及3個首次於天然物中分離獲得之已知化合物N-benzyl-S-methyl-thiocarbamate (6)、ethyl benzylcarbamate (7)和(S)-2-benzyl-3-thioxohexahydropyrrolo-[1,2-c]-imidazole-1-one (8)。 分離得到的化合物結構皆利用核磁共振圖譜1D NMR (1H NMR和13C NMR)及2D NMR (COSY、HSQC、HMBC、NOESY)進行解析，加上各種物理數據分析，包括熔點、紅外線吸收光譜、紫外線吸收光譜及高解析電灑式質譜儀，並經由與化學結構資料庫(Reaxys)進行相關文獻比對而得到確認。另外化合物1為結晶化合物，其結構更經由X-ray單晶繞射解析進行更進一步的證明。 最後將所分離得到的化合物分別進行抗發炎以及抗癌細胞的生物活性檢測，結果發現化合物5和7對FMLP/CB的引起人類嗜中性白血球發炎反應後之superoxide陰離子釋出有抑制的效果，其IC50值分別為0.35 和2.40 μg/mL，化合物7對elastase的釋出也具有抑制的效果，其IC50值為4.99 μg/mL。

In this thesis, the phytochemical studies of the chemical constituents of Bretschneidera sinensis Hemsley was investigated. The ethanol extracts of the twigs, leaves and fruit shells of B. sinensis were isolated by using extensive column chromatography, which led to eight compounds including five new sulfur-containing alkaloids. Compounds 1─3 were identified as new heterocyclic compounds which belong to the skeleton of 3-methyl-1,3-benzothiazine-2,4-dione, while compounds 4 and 5 were characterized as new sulfur-containing alkaloids which belong to the skeleton of N-benzyl-S-methyl- thiocarbamate. In addition, three known compounds, N-benzyl-S-methylthiocarbamate (6), ethyl benzylcarbamate (7) and (S)-2-benzyl-3-thioxohexahydropyrrolo[1,2-c]imidazole-1-one (8) were obtained from the natural source for the first time. The above structures were elucidated and established on the basis of spectroscopic analysis such as 1D NMR (1H and 13C NMR) and 2D NMR (COSY, HSQC, HMBC and NOESY) techniques, as well as the physical methods including melting point, infrared spectroscopy (IR), ultraviolet spectroscopy (UV) and high resolution electrospray ionization mass spectroscopy (HRESIMS). Besides, the structures of all the isolated compounds were further confirmed by comparing with the chemical databases, Reaxys. In addition, the structure of compound 1 was further confirmed by single-crystal X-ray diffraction analysis. Finally, all the isolated compounds were tested and evaluated for the biological activities including anti-inflammatory and cytotoxic activities. The results showed that compounds 5 and 7 inhibited the superoxide anion generation release in FMLP/CD-induced human neutrophils with IC50 of 0.35 and 2.40 μg/mL, respectively. Compound 7 also inhibited the elastase release in FMLP/CB-induced human neutrophils with IC50 of 4.99 μg/mL.