第一部分. 中國穿鞘花全草化學成分與抗結核活性之研究
第二部分. 披覆炭團菌二次代謝產物與抗發炎活性之研究

Part I: 中國穿鞘花 (Amischotolype hispida (Less.＆ A. Rich.) Hong)為鴨跖草科 (Commelinaceae)植物，為多年生草本，分布於全島中低海拔之森林中。穿鞘花屬世界有20種，而台灣僅一種，而本植物全草之甲醇萃取物具有抗結核活性，且過去對於穿鞘花屬未曾有天然物分離及其相關活性之研究。由乙酸乙酯可溶部具抗結核活性層利用活性導向劃分法得到共25個化合物，包含四個新化合物: amisbenzoic acid (1)、amisnolide (2)、amisphytin (3)與amislignol (4)及一個由天然界首次分離之lignan類化合物: (–)-glaberide I (5)，以及20個已知化合物: veratic acid (6)、4-methoxybenzoic acid (7)、(–)-syringaresinol (8)、(±)-pinoresinol (9)、(+)-4-ketopinoresinol (10)、methyl palmitate (11)、methyl linoleate (12)、palmitic acid (13)、α-tocopherol (14)、γ-tocopherol (15)、δ-tocopherol (16)、ubiquinone Q9 (17)、α-tocopheryl quinone (18)、trans-phytol (19)、indole-3-carbaldehyde (20)、24-methylenecycloartanol (21)、cycloartenol (22)、β-sitosterol (23)、β-sitostenone (24)與ergosterol peroxide (25)。上述化合物皆由各種圖譜分析決定結構。抗結核活性實驗中，palmitic acid (13)顯示有抗結核活性，其MIC為20.0 μg/mL，並也針對了四種不同市售長鏈類似物進行抗結核活性探討，其中比palmitic acid少了兩個碳的myristic acid，顯示較弱的抗結核活性，其MIC為50.0 μg/mL，因此推測其鏈長為14與16個碳，對於抗結核之活性是有其意義存在。 Part II: 披覆炭團菌 (Hypoxylon investiens)為台灣固有種植物之內生真菌，是從樟科竹頭角木薑子之莖部所分離出來，並以液態發酵培養之正丁醇粹取物以管柱層析法進行分離得到共29個化合物，包含三個新的化合物: hypoxyloamide (26)、8-methoxynaphthalene-1,7-diol (27)及hypoxylonol (28)，七個化合物由天然物首次分離: investiamide (29)、hypoxypropanamide (30)、hypoxylonol A (31)、investienol (32)、2-heptylfuran (33)、(S)-5-methyl-8-O-methylmellein (34)及(R)-O-methylsclerone (35)，以及19個已知化合物: indole-3-carbaldehyde (20)、ergosterol peroxide (25)、N-acetyltyramine (36)、nicotinamide (37)、8-methoxynaphthalen-1-ol (38)、1,8-dimethoxynaphthalene (39)、xylaranol B (40)、epi-guaidiol A (41)、(R)-5-hydroxymethylmellein (42)、(R)-8-O-methylmellein (43)、(R)-5-methylmellein (44)、(–)-regiolone (45)、xylarenone (46)、2-hydroxyphenethyl alcohol (47)、cyclo(L-Pro-L-Tyr) (48)、monascuspyrrole (49)、3-hydroxy-2-methyl-4H-pyran-4-one (50)、4-hydroxy-6-methyl-2H-pyran-2-one (51)與ergosta-4,6,8(14),22-tetraen-3-one (52)。上述化合物之結構皆由各種圖譜分析決定。將足夠量之化合物進行抗發炎功效之篩選，測試在RAW 264.7細胞株中對於抑制脂多醣誘導之NO、IL-6及TNF-α所產生之能力。其中，8-methoxynaphthalene1,7-diol (27)、8-methoxynaphthalen-1-ol (38) 及1,8-dimethoxynaphthalene (39)具有強的抑制NO產生之活性，其IC50分別為11.8, 17.8及13.3 μM，且優於對照組quercetin (IC50 36.8 μM)。化合物27、38及39也顯示了強的抑制IL-6產生之活性，其IC50分別為9.2, 18.0及2.0 μM，且優於對照組quercetin (IC50 31.3 μM)。

關鍵字

中國穿鞘花；鴨跖草科；抗結核病；披覆炭團菌；炭角菌科；內生菌；竹頭角木薑子；抗發炎

並列摘要

Part I: Amischotolype hispida (Less.＆ A. Rich.) Hong (Commelinaceae) is a perennial herb, distributed over forest floors at low to middle altitudes throughout Taiwan. There are 20 species of Amischotolype genus around the world and only one species in Taiwan. The methanolic extract of the whole plant of this plant was shown with antitubercular activity. However, the phytochemistry and biological activities of Amischotolype genus have never been conducted. Bioassay-guided fractionation of the active ethyl acetate-soluble layer has led to the isolation of four new compounds: amisbenzoic acid (1), amisnolide (2), amisphytin (3), amislignol (4) and one compound first isolated from nature, (–)-glaberide I (5), along with 20 known compounds, veratic acid (6), 4-methoxybenzoic acid (7), (–)-syringaresinol (8), (±)-pinoresinol (9), (+)-4-ketopinoresinol (10), methyl palmitate (11), methyl linoleate (12), palmitic acid (13), α-tocopherol (14), γ-tocopherol (15), δ-tocopherol (16), ubiquinone Q9 (17), α-tocopheryl quinone (18), trans-phytol (19), indole-3-carbaldehyde (20), 24-methylenecycloartanol (21), cycloartenol (22), β-sitosterol (23), β-sitostenone (24), and ergosterol peroxide (25). The structures of isolates were determined by spectroscopic techniques. Palmitic acid (13) showed antitubercular activity with MIC value of 20.0 μg/mL and other four different lengths of fatty acid analogues such as myristic acid with less two methylenes than palmitic acid which showed weak antitubercular activity with MIC value of 50.0 μg/mL. From the biological results of the study, it is meaningful for antitubercular activity on the carbon numbers from 14 and 16 of saturated fatty acid analogues. Part II: Hypoxylon investiens was an endophytic fungus, isolated from the stem of Taiwan endemic plant, Litsea akoensis Hayata var. chitouchiaoensis Liao (Lauraceae). H. investiens was processed through liquid fermentation. The n-BuOH extract of H. investiens was further chromatographed and led to the isolation of three new compounds, hypoxyloamide (26), 8-methoxynaphthalene-1,7-diol (27), and hypoxylonol (28), and ten compounds first isolated from nature, investiamide (29), hypoxypropanamide (30), hypoxylonol A (31), investienol (32), 2-heptylfuran (33), (S)-5-methyl-8-O-methylmellein (34), and (R)-O-methylsclerone (35), along with 19 known compounds, indole-3-carbaldehyde (20), ergosterol peroxide (25), N-acetyltyramine (36), nicotinamide (37), 8-methoxynaphthalen-1-ol (38), 1,8-dimethoxynaphthalene (39), xylaranol B (40), epi-guaidiol A (41), (R)-5-hydroxymethylmellein (42), (R)-8-O-methylmellein (43), (R)-5-methylmellein (44), (–)-regiolone (45), xylarenone (46), 2-hydroxyphenethyl alcohol (47), cyclo(L-Pro-L-Tyr) (48), monascuspyrrole (49), 3-hydroxy-2-methyl-4H-pyran-4-one (50), 4-hydroxy-6-methyl-2H-pyran-2-one (51), and ergosta-4,6,8(14),22-tetraen-3-one (52). The structures of isolates were determined by spectroscopic techniques. The isolates with enough amounts were screened for their ability to inhibit NO, IL-6, and TNF-α production in LPS-activated RAW 264.7 cells. Among these isolates, 8-methoxynaphthalene-1,7-diol (27), 8-methoxynaphthalen-1-ol (38) and 1,8-dimethoxynaphthalene (39) showed the stronger NO inhibitory activity with IC50 values of 11.8, 17.8, and 13.3 μM than the positive control quercetin (IC50= 36.8 μM). Compounds 27, 38, and 39 also showed stronger IL-6 inhibitory activity with IC50 values of 9.2, 18.0, and 2.0 μM than the positive control quercetin (IC50= 31.3 μM).

並列關鍵字

Amischotolype hispida ； Commelinaceae ； anti-tubercular activity ； Hypoxylon investiens ； Xylariaceae ； endophytic fungus ； Litsea akoensis var. chitouchiaoensis ； Lauraceae ； anti-inflammatory

參考文獻

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