Antitubercular Activity and Inhibitory Effect on Epstein-Barr Virus Activation of Sterols and Polyisoprenepolyols from an Edible Mushroom, Hypsizigus marmoreus

Abstract

Hypsizigus marmoreus (Buna-shimeji) has recently become a popular edible mushroom in Asia, and has been cultivated by culture methods such as bottle culture and bag-culture. It has been reported to exhibit many biological activities such as anti-tumor, 1,2) antioxidant, 1) and antiproliferative activity. 3) Several homologous polyisoprenepolyols named hypsiziprenols have recently been characterized in this mushroom. 4) Hypsiziprenols are the possible biosynthetic precursors of gymnopilins, bitter principles of H. marmoreus. 4) In the course of our studies on the bioactive principles of natural medicines and foodstuffs, 5,6) we evaluated the constituents of H. marmoreus for their antitubercular activity 7) as well as their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). 5,6) In this paper, we describe the antitubercular activity against Mycobacterium tuberculosis of seven sterols (1-7) and eight polyisoprenepolyols, hypsiziprenols (8-15), and inhibitory effects on the induction of EBV-EA by the TPA of seven sterols (1-7) and three hypsiziprenols (8, 10, 12) isolated from the non-saponifiable lipid (NSL) fraction of the extract of H. marmoreus. MATERIALS AND METHODSExtraction, Isolation, and Identification Sliced fresh bodies of Hypsizigus marmoreus (11 kg), cultivated in Nagano Prefecture in 2001, were air-dried. The dried mushroom (833 g) was then extracted with hot CH 2 Cl 2 to give an extract (86 g). Alkaline hydrolysis (5% KOH in methanol (MeOH), reflux, 3 h) of the extract yielded a neutral nonsaponifiable lipid (NSL) fraction (9.0 g). Crystallization of the NSL fraction from MeOH yielded a precipitate (2.8 g), which was identified as ergosterol (1), 2) and a filtrate (4.9 g).The filtrate was subjected to column chromatography on silica gel (150 g) using an n-hexane-ethyl acetate (EtOAc) gradient of 1 : 0-1 : 1 as an eluent, which yielded fractions containing monohydroxy sterols (fraction A; Rf 0.83 on TLC developed with EtOAc-MeOH, 4 : 1), polyhydroxy sterols (fraction B; Rf 0.48), and polyisoprenepolyols (fraction C; Rf 0.24). Fractions A-C were purified by further chromatography on silica gel. Reversed-phase preparative HPLC (HPLC I; 25 cmϫ10 mm i.d., Pegasil ODS II (Senshu Scientific Co., Ltd., Tokyo, Japan) column; mobile phase: MeOH-H 2 O- Farmers' Federation; 787-1 Suzaka, Sazaka, Nagano 382-0084, Japan: and e College of Pharmacy, Nihon University; 7-7-1 Narashinodai, Funabashi, Chiba 274-8555, Japan. Received January 18, 2005; accepted February 28, 2005 Seven sterols (1-7) and eight polyisoprenepolyols (8-15), isolated from the non-saponifiable lipid fraction of the dichloromethane extract of an edible mushroom, Hypsizigus marmoreus (Buna-shimeji), were tested for their antitubercular activity against Mycobacterium tuberculosis strain H 37 Rv using the Microplate Alamar Blue Assay (MABA). Six sterols (2-7) and two polyisoprenepolyols (8, 12) showed a minimum inhibitory concentration (MIC) in the range of 1-51 m mg...

Isolation of ergosterol peroxide from Nomuraea rileyi infected larvae of tobacco cutworm

“…Additional biological activities of ergosterol peroxide have been reported, i.e., antioxidant (Kim et al 1999), antimycobacterial (Cantrell et al 1999;Akihisa et al 2005;Duarte et al 2007), antiplasmodial (Kuria et al 2002), leishmanicidal activity (Correa et al 2006), antiinflammatory (Yasukawa et al 1996;Gao et al 2007;Kobori et al 2007), immunosuppressive (Kuo et al 2003), anti-atherosclerosis (Kim et al 2005) and aldose-reductase inhibitory activity (Lee et al 2006). Aside from the reported biological activities, we found that ergosterol peroxide had an effect on in vitro insect cells and an insecticidal activity against 3rd instar S. litura larvae via topical application.…”

“…and Penicillium sp. (Nemec et al 1997;Kuo et al 2005), mushrooms (Mizushina et al 1998;Kim et al 1999;Li et al 2004;Akihisa et al 2005;Gao et al 2007), marine organisms (Gunatilaka et al 1981), lichens (Rubio et al 2005), algae (Sheffer et al 1986;Yasukawa et al 1996) and plants (Kuria et al 2002;Kim et al 2005;Macias et al 2006). This is the first report of ergosterol peroxide isolation from N. rileyi.…”

Anti‐Inflammatory and Anti‐Tumor‐Promoting Effects of Triterpene Acids and Sterols from the Fungus Ganoderma lucidum

“…-The MeOH extract of the fruiting bodies of G. lucidum was purified by column chromatography on silica gel followed by preparative reversephase HPLC, yielding four known lucidenic acid derivatives, lucidenic acid B (3) [8], methyl lucidenate C (4) [9], lucidenic acid F (9) [6] [10], and lucidenic acid N (10) [11], six known ganoderic acids and related compounds, ganoderic acid A (20) [12], ganoderic acid C 1 (21) [10], ganoderic acid C 2 (22) [10], ganoderic acid DM (24) [13], ganodermanondiol (21) [14], and ganolactone (27) [15], as well as six known sterols, fungisterol (28) [16], 5,6-dihydroergosterol (29) [16], ergosterol (30) [17], ergosterol peroxide (31) [17], 9(11)-dehydroergosterol peroxide (32) [18], and demethylincisterol A 3 (33) [19] [20]. These compounds were identified by comparison of the spectroscopic data with those reported in the literature.…”

Antitubercular Activity of Mushrooms (Basidiomycetes) and their Metabolites

“…Two other derivatives of vulpinic acid, (6) and 7, obtained from 5 by semi-synthetic modifications showed very weak antitubercular activity (MIC of 100 μg/mL) [10]. Recently, several secondary metabolites isolated from the edible mushroom Hypsizigus marmoreus have been shown to exhibit antitubercular activity and an inhibitory effect on tumor promotion [12]. Hypsizigus marmoreus (Peck) Bigel., known in Asia under a common name buna-shimeji, has been reported to exhibit many biological activities such as anti-tumor, antioxidant and antiproliferative activity.…”

“…Six sterols (9)(10)(11)(12)(13)(14) and two polyisoprenepolyols (15,19) showed an MIC in the range of 1-51 μg/mL, while the other compounds (8, 16-18, 20-22) were inactive (MIC > 128 μg/mL). Ergosterol peroxide (9) and cerevisterol (10) showed the highest antitubercular potency with MIC values of 1 μg/mL and 6 μg/mL respectively, as compared to a positive control (rifampin MIC 0.25 μg/mL).…”