重緣葉馬尾藻衍生真菌株之化學活性成分研究

Abstract

本研究自海洋褐藻重緣葉馬尾藻 (Sargassum cristaefolium C. Agardh) 中分離出真菌株Penicillium sumatraense SC29、Xylaria acuta SC1019、Scytalidium lignicola SC228和Hypomontagnella monticulosa SC1047，經液態及固態發酵後以有機溶劑進行分配萃取發酵產物，再將粗萃物以開放式管柱和高效液相層析儀 (HPLC) 進行分析、分離與純化，所得純質以核磁共振光譜 (NMR) 鑑定分子結構，再透過質譜、紅外光譜、紫外光譜、圓二色光譜、旋光度及Mosher’s酯化反應等數據佐證結構的正確性。計得到23個未曾報導的聚酮類化合物，包含 penisterine A (PS-1)、penisterines C-E (PS-3~PS-5)、xylarilactone A (XA-1)、xylarilactone B (XA-2)、xylarilactone C (XA-3)、ent-gedebic acid 8-O-α-D-glucopyranoside (XA-4)、5R-hydroxylmethylmellein 11-O-α-D- glucopyranoside (XA-5)、ent-hymatoxin E 16-O-α-D-mannopyranoside (XA-6)、19,20-epoxycytochalasin S (XA-7)、19,20-epoxycytochalasin T (XA-8)、(2R)-butylitaconic acid (XA-9)、scytabenzofurans A-C (SL-1~SL-3)、(3S,4S)-5-chloro-3,4-dihydro-4,6,8-trihydroxy-3-methyl-1H-2-benzopyran-1-one (SL-4)、hypoketides A-F (HM-1~HM-6)，一個天然物首次分離化合物penisterine B (PS-2)，及21個已知化合物。在生物活性方面，penisterine D (PS-4)、19,20-epoxycytochalasin S (XA-7)、19,20-epoxycytochalasin T (XA-8)、cytochalasin C (XA-21)、(-)-mycorrhizin A (SL-5)、(-)-dechloromycorrhizin A (SL-6)、(-)-trans-3,4-dihydro-2,4,8-trihydroxynaphthalen-1(2H)-one (SL-7)、hypoketide A (HM-1)、hypoketide B (HM-2) 和sporothriolide (HM-7) 表現出對EPC生長的抑製作用，IC50值分別為28.5 ± 2.2、0.4 ± 0.1、0.5 ± 0.1、0.5 ± 0.1、30.6 ± 0.8、0.5 ± 0.1、42.7 ± 0.9、47.7 ± 5.5、21.6 ± 0.6和63.8 ± 1.8 µM。Xylarilactone A (XA-1)、5R-hydroxylmethylmellein 11-O-α-D-glucopyranoside (XA-5)、19,20-epoxycytochalasin S (XA-7)、PC-2 (XA-10)、5,6-dihydro-4-methoxy-6-(pentanoyloxy)-2H-pyran-2-one (XA-17)、scytabenzofurans A-C (SL-1~SL-3)、(3S,4S)-5-chloro-3,4-dihydro- 4,6,8-trihydroxy-3-methyl-1H-2-benzopyran-1-one (SL-4)、(-)-trans-3,4-dihydro- 2,4,8-trihydroxynaphthalen-1(2H)-one (SL-7)、(R)-6-hydroxymellein (SL-8)、hypoketide B (HM-2) 和sporothriolide (HM-7) 表現出對BV-2細胞產生的NO具有抑制作用，IC50值分別為19.6 ± 0.4、16.1 ± 0.6、11.4 ± 0.3、15.2 ± 0.9、11.8 ± 0.5、49.0 ± 1.2、23.4 ± 0.4、34.2 ± 0.5、22.7 ± 1.5、44.8 ± 2.6、19.6 ± 0.1、4.9 ± 0.3和8.9 ± 0.1 µM。

In this study, fungal strains Penicillium sumatraense SC29, Xylaria acuta SC1019, Scytalidium lignicola SC228, and Hypomontagnella monticulosa SC1047 were isolated and identified from a marine brown algae Sargassum cristaefolium. Sephadex LH-20 open column separation followed by HPLC purification of liquid- and solid-state fermented products of the fungal strains were carried out, the structures were elucidated by spectroscopic analysis including NMR, MS, IR, UV, CD, optical rotational and Mosher’s method. This analysis led to identification of 23 new poketides, including penisterine A (PS-1), penisterines C-E (PS-3~PS-5), xylarilactones A-C (XA-1~XA-3), ent-gedebic acid 8-O-α-D-glucopyranoside (XA-4), 5R-hydroxylmethylmellein 11-O-α-D-glucopyranoside (XA-5), ent-hymatoxin E 16-O-α-D-mannopyranoside (XA-6), 19,20-epoxycytochalasin S (XA-7), 19,20-epoxycytochalasin T (XA-8), (2R)-butylitaconic acid (XA-9), scytabenzofurans A-C (SL-1~SL-3), (3S,4S)-5-chloro-3,4-dihydro-4,6,8-trihydroxy-3-methyl-1H-2-benzopyran-1-one (SL-4), and hypoketides A-F (HM-1~HM-6), along with one compound firstly isolated from nature, and 21 known compounds. Of these, penisterine D (PS-4), 19,20-epoxycytochalasin S (XA-7), 19,20-epoxycytochalasin T (XA-8), cytochalasin C (XA-21), (-)-mycorrhizin A (SL-5), (-)-dechloromycorrhizin A (SL-6), (-)-trans-3,4-dihydro-2,4,8-trihydroxynaphthalen-1(2H)-one (SL-7), hypoketide A (HM-1), hypoketide B (HM-2), and sporothriolide (HM-7) displayed anti-angiogenic activities by suppressing the growth of human endothelial progenitor cells with IC50 values of 28.5 ± 2.2, 0.4 ± 0.1, 0.5 ± 0.1, 0.5 ± 0.1, 30.6 ± 0.8, 0.5 ± 0.1, 42.7 ± 0.9, 47.7 ± 5.5, 21.6 ± 0.6, and 63.8 ± 1.8 µM, respectively. Xylarilactone A (XA-1), 5R-hydroxylmethylmellein 11-O-α-D-glucopyranoside (XA-5), 19,20-epoxycytochalasin S (XA-7), PC-2 (XA-10), 5,6-dihydro-4-methoxy-6-(pentanoyloxy)-2H-pyran-2-one (XA-17), scytabenzofurans A-C (SL-1~SL-3), (3S,4S)-5-chloro-3,4-dihydro-4,6,8-trihydroxy- 3-methyl-1H-2-benzopyran-1-one (SL-4), (-)-trans-3,4-dihydro-2,4,8-trihydroxynaphthalen- 1(2H)-one (SL-7), (R)-6-hydroxymellein (SL-8), hypoketide B (HM-2), and sporothriolide (HM-7) showed nitric oxide production inhibitory activities in lipopolysaccharide-activated BV-2 microglial cells with IC50 values of 19.6 ± 0.4, 16.1 ± 0.6, 11.4 ± 0.3, 15.2 ± 0.9, 11.8 ± 0.5, 49.0 ± 1.2, 23.4 ± 0.4, 34.2 ± 0.5, 22.7 ± 1.5, 44.8 ± 2.6, 19.6 ± 0.1, 4.9 ± 0.3, and 8.9 ± 0.1 µM.