防葵衍生真菌株Diaporthe ueckerae NTU1943之成分研究

Abstract

本研究自濱海植物防葵 (Peucedanum japonicum Thumb.) 中分離出真菌株Diaporthe ueckerae NTU1943，接著使用半淡海水馬鈴薯提取物葡萄糖培養基 (potato dextrose broth, PDB) 液態醱酵14天，以及以糙米進行固態醱酵30天，之後以乙酸乙酯進行分配萃取醱酵產物，再針對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 (MS)、紅外線光譜 (IR)、紫外光譜 (UV) 及旋光度 (optical rotation) 等數據佐證結構的正確性。計得到2個倍半萜 (sesquiterpenes)、1個單萜 (monoterpene)、2個苯乙烷類 (phenylethanoids)、6個δ-內酯類 (δ-lactones)、3個呫噸酮二聚物 (xanthone dimers) 和1個縮酚肽 (depsipeptide)，分別為altiloxin F (1)、altiloxin A (2)、p-menthane-1,2,4-triol (3)、phenylacetic acid (4)、tyrosol (5)、(3R)-5-hydroxymellein (6)、mellein (7)、(3R,4R)-4-hydroxymellein (8)、(3R,4S)-4-hydroxymellein (9)、(S)-8-O-methylmellein (10)、nectriapyrone (11)、diaporxanthone F (12)、dicerandrols A和B (13-14) 與fusaristatin A (15)，其中altiloxin F (1) 為未曾報導之新化合物。在抗病毒與抗菌活性測試上，顯示化合物8在4123 μM濃度下，對於貓傳染性腹膜炎病毒 (feline infectious peritonitis virus, FIPV) 具有弱的抗病毒活性；在抗水產致病菌評估中，所得純質皆無抗菌活性；化合物13在10 μM濃度下對EBV (Epstein-Barr virus) 抑制效果顯著，但同時對P3HR1細胞具有細胞毒性，且在20 μM濃度下對LPS誘導BV-2 細胞之一氧化氮 (NO) 具有明顯的抑制效果，其抑制率為113.7%，但觀察到其細胞存活率僅為9.3%，推測其NO產生的抑制效果同樣可能來自對細胞的毒殺活性。

In this study, a fungal strain Diaporthe ueckerae NTU1943 was isolated and identified from a littoral plant Peucedanum japonicum Thumb.. Column separation followed by HPLC purification of liquid-state and solid-state fermented products of the fungal strain was carried out, which has led to the isolation of fifteen compounds 1–15. Their structures were elucidated by spectroscopic analysis including NMR, MS, IR, UV, and optical rotational data to be two sesquiterpenes, one monoterpene, two phenylethanoids, six δ-lactones, three xanthone dimers, and one depsipeptide, and were named altiloxin F (1), altiloxin A (2), p-menthane-1,2,4-triol (3), phenylacetic acid (4), tyrosol (5), (3R)-5-hydroxymellein (6), mellein (7), (3R,4R)-4-hydroxymellein (8), (3R,4S)-4-hydroxymellein (9), (S)-8-O-methylmellein (10), nectriapyrone (11), diaporxanthone F (12), dicerandrols A and B (13 and 14), and fusaristatin A (15). Of these, 1 was a previously unreported chemical entity. Compound 8 exhibited slight antiviral activity against feline infectious peritonitis virus (FIPV) at a concentration of 4123 μM. All compounds showed no significant antibacterial activity. Dicerandrol A (13) exhibited significant Epstein-Barr virus (EBV) inhibitory activity, but it was possibly caused by its cytotoxicity in P3HR1 cells at a concentration of 10 μM. Dicerandrol A (13) showed significant NO production inhibitory activity, but it was possibly caused by its cytotoxicity in BV-2 cells at a concentration of 20 μM.