The Effects of Ergosta-7,9(11),22-trien-3ß-ol from Antrodia camphorata on the Biochemical Profile and Exercise Performance of Mice.

Antrodia camphorata (AC) is a rare and unique mushroom that is difficult to cultivate. Previous studies have demonstrated the bioactivity of the compound Ergosta-7,9(11),22-trien-3ß-ol (EK100) from AC in submerged culture. The purpose of this study is to evaluate the potential beneficial effects of EK100 on fatigue and ergogenic functions following physiological challenge. Male ICR (Institute of Cancer Research) mice were randomly divided into three groups (n = 8 per group) and orally administered EK100 for six weeks at 0 (Vehicle), 10 (EK100-1X), and 20 (EK100-2X) mg/kg/day. The six-week Ek100 supplementation significantly increased grip strength (P = 0.0051) in trend analysis. Anti-fatigue activity was evaluated using 15-min. acute exercise testing and measuring the levels of serum lactate, ammonia, glucose, blood urea nitrogen (BUN), and creatine kinase (CK) after a 15-min. swimming exercise. Our results indicate that AC supplementation leads to a dose-dependent decrease in serum lactate, ammonia, BUN, and CK activity after exercise and significantly increases serum glucose and glycogen content in liver tissues. Biochemical and histopathological data demonstrated that long term daily administration of EK100 for over six weeks (subacute toxicity) was safe. EK100's anti-fatigue properties appear to be through the preservation of energy storage, increasing blood glucose and liver glycogen content, and decreasing the serum levels of lactate, ammonia, BUN, and CK. EK100 could potentially be used to improve exercise physiological adaptation, promote health, and as a potential ergogenic aid in combination with different nutrient strategies.

Ergosterol peroxide from marine fungus Phoma sp. induces ROS-dependent apoptosis and autophagy in human lung adenocarcinoma cells.

As part of our ongoing search for novel therapeutic structures from microorganism, the chemical examination of marine fungus Phoma sp. resulted in the isolation of ergosterol, ergosterol peroxide (EP), and 9,11-dehydroergosterol peroxide (DEP). The bioassay results demonstrated that the three isolates reduced the viability of various cancer cells, with EP being highest in human lung cancer cell line A549 cells. EP induced caspase-dependent apoptosis through mitochondrial damage in A549 cells. Additionally, EP-induced ROS generation and apoptosis were attenuated by ROS-generating enzymes inhibitors and antioxidant N-acetylcysteine, indicated that ROS played an important role in EP-mediated apoptosis in A549 cells. Furthermore, it was observed that EP induced ROS-dependent autophagy, which attenuated apoptosis in A549 cells. On the other hand, EP reduced the LPS/ATP-induced proliferation and migration of A549 cells through attenuated NLRP3 inflammasome activity. Additionally, EP showed synergistic cytotoxic effect with antitumor drug Sorafenib in A549 cell viability inhibition. Furthermore, Micro-Western Array and Western blot analyses demonstrated that the protein levels of EGFR, HSP27, MEK5, AKT1, mTOR, Smad2, Smad3, TAB1, NF-κB, and HIF1-α decreased, while the levels of p-p38α, p-ERK1/2, p-JNK, fibronectin and p27 increased. Collectively, the results of this study demonstrated that EP might be useful to develop a therapeutic candidate for lung cancer complications.

Hyfraxins A and B, cytotoxic ergostane-type steroid and lanostane triterpenoid glycosides from the invasive ash dieback ascomycete Hymenoscyphus fraxineus.

A virulent culture of Hymenoscyphus fraxineus, the causal agent of ash dieback, was investigated for its production of secondary metabolites in a 70 L batch fermentation. Chemical analysis of the mycelial extract by means of flash chromatography and preparative HPLC led to the isolation of a new ergostane-type steroid (1) and a new related lanostane triterpenoid (2), both revealing the same glycosylation pattern. While their planar structures were elucidated by HR-ESIMS and NMR data, relative stereochemistry was assigned by ROESY correlations in conjunction with H,H and C,H coupling constants. Absolute configuration was determined based on ROESY correlations between the aglycons and the sugar moieties, which were identified in both cases as d-mannose by GC/MS analysis of the trimethylsilylated derivatives. The isolated compounds, for which we propose the trivial names hyfraxins A (1) and B (2), were found to be cytotoxic against the mouse fibroblast cell line L929 and exhibited moderate to weak activity against Gram-positive bacteria.

Lethality of cytochalasin B and other compounds isolated from fungus Aspergillus sp. (Trichocomaceae) endophyte of Bauhinia guianensis (Fabaceae).

Endophytic fungi are fungi that colonize internal tissues of plants; several biologically active compounds have been isolated from these fungi. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, this study aimed the isolation and structural identification of ergosterol (1), ergosterol peroxide (2), mevalonolactone (3), cytochalasin B (4) and cytochalasin H (5) from Aspergillus sp. EJC 04, an endophytic fungus from Bauhinia guianensis. The cytochalasin B (4) and the diacetate derivative of cytochalasin B (4a) showed high lethality in the brine shrimp assay. This is the first occurrence of cytochalasins in Amazonian endophytic fungi from B. guianensis.

Antimicrobial ergosteroids and pyrrole derivatives from halotolerant Aspergillus flocculosus PT05-1 cultured in a hypersaline medium.

In order to obtain more structurally novel and bioactive lead compounds for subsequent drug discovery, we have shifted the focus of our study from traditional microbial resources to 'extremophiles'. In this study, a halotolerant fungus Aspergillus flocculosus PT05-1 was isolated from the sediment of Putian saltern of Fujian Province of China in a hypersaline medium. Two new compounds, (22R,23S)-epoxy-3ß,11α,14ß,16ß-tetrahydroxyergosta-5,7-dien-12-one (1) and 6-(1H-pyrrol-2-yl)hexa-1,3,5-trienyl-4-methoxy-2H-pyran-2-one (5) (existed as a pair of epimers with the configuration of 1E,3Z,5E and 1E,3E,5E separately), along with nine known compounds were isolated and identified from the fermentation broth of A. flocculosus PT05-1 grown at a 10 % saline medium. New ergosteroid 1 together with 7-nor-ergosterolide (2) and 3ß-hydroxyergosta-8,24(28)-dien-7-one (3) showed cytotoxicity against HL-60 and BEL-7402 cells with IC50 values of 12-18 µM, and antimicrobial activity against Enterobacter aerogenes, Pseudomonas aeruginosa, and Candida albicans with MIC values of 1.6-15 µM, respectively. New compound 5 exhibited antibacterial effect on E. aerogenes with MIC value of 3.7 µM. This study also showed great prospects in developing medicinal resources from extremophiles.

Chemical constituents of Euphorbia tangutica.

Sixteen known compounds isolated from the whole plants of Euphorbia tangutica, including phorbol-13-actate (1) previously synthesised and obtained from a natural source for the first time, were evaluated in vitro against a panel of human cancer cell lines using the MTT method. Among them, ergosterol (6) exhibited significant cytotoxic activity against HL-60 cell line with an IC(50) value of 3.3 µM, and 3ß,5α-dihydroxy-15ß-cinnamoyloxy-14-oxolathyra-6 Z,12 E-diene (7) also displayed moderate activity.

Chemical constituents of Papulaspora immersa, an endophyte from Smallanthus sonchifolius (Asteraceae), and their cytotoxic activity.

Papulaspora immersa H. H. Hotson was isolated from roots and leaves of Smallanthus sonchifolius (Poepp. and Endl.) H. Rob. (Asteraceae), traditionally known as Yacon. The fungus was cultured in rice, and, from the AcOEt fraction, 14 compounds were isolated. Among them, (22E,24R)-8,14-epoxyergosta-4,22-diene-3,6-dione (4), 2,3-epoxy-1,2,3,4-tetrahydronaphthalene-c-1,c-4,8-triol (10), and the chromone papulasporin (13) were new secondary metabolites. The spectral data of the known natural products were compared with the literature data, and their structures were established as the (24R)-stigmast-4-en-3-one (1), 24-methylenecycloartan-3ß-ol (2), (22E,24R)-ergosta-4,6,8(14),22-tetraen-3-one (3), (-)-(3R,4R)-4-hydroxymellein (5), (-)-(3R)-5-hydroxymellein (6), 6,8-dihydroxy-3-methylisocoumarin (7), (-)-(4S)-4,8-dihydroxy-α-tetralone (8), naphthalene-1,8-diol (9), 6,7,8-trihydroxy-3-methylisocoumarin (11), 7-hydroxy-2,5-dimethylchromone (12), and tyrosol (14). Compound 4 showed the highest cytotoxic activity against the human tumor cell lines MDA-MB435 (melanoma), HCT-8 (colon), SF295 (glioblastoma), and HL-60 (promyelocytic leukemia), with IC50 values of 3.3, 14.7, 5.0 and 1.6 µM, respectively. Strong synergistic effects were also observed with compound 5 and some of the isolated steroidal compounds.

Chemical examination of the Sponge Phycopsis sp.

Two new compounds 7alpha,8alpha-epoxy theonellin isothiocyanate (1) and 5alpha,8alpha-epidioxyergosta-6Z,22Z,25-trien-3beta-ol (2) along with two known compounds, theonellin isothiocyanate (3) and theonellin formamide (4) have been isolated from the sponge Phycopsis sp. Compound 2 showed cytotoxic activity against HL-60 and U937 human cancer cell lines with IC(50) values of 5.96+/-0.02 and 31.72+/-0.55 microg/ml, respectively.

Withanolides, a new class of natural cholinesterase inhibitors with calcium antagonistic properties.

The withanolides 1-3 and 4-5 isolated from Ajuga bracteosa and Withania somnifera, respectively, inhibited acetylcholinesterase (AChE, EC 3.1.1.7) and butyrylcholinesterase (BChE, EC 3.1.1.8) enzymes in a concentration-dependent fashion with IC50 values ranging between 20.5 and 49,2 microm and 29.0 and 85.2 microm for AChE and BChE, respectively. Lineweaver-Burk as well as Dixon plots and their secondary replots indicated that compounds 1, 3, and 5 are the linear mixed-type inhibitors of AChE, while 2 and 4 are non-competitive inhibitors of AChE with K(i) values ranging between 20.0 and 45.0 microm. All compounds were found to be non-competitive inhibitors of BChE with K(i) values ranging between 27.7 and 90.6 microm. Molecular docking study revealed that all the ligands are completely buried inside the aromatic gorge of AChE, while compounds 1, 3, and 5 extend up to the catalytic triad. A comparison of the docking results showed that all ligands generally adopt the same binding mode and lie parallel to the surface of the gorge. The superposition of the docked structures demonstrated that the non-flexible skeleton of the ligands always penetrates the aromatic gorge through the six-membered ring A, allowing their simultaneous interaction with more than one subsite of the active center. The affinity of ligands with AChE was found to be the cumulative effects of number of hydrophobic contacts and hydrogen bonding. Furthermore, all compounds also displayed dose-dependent (0.005-1.0 mg/mL) spasmolytic and Ca2+ antagonistic potentials in isolated rabbit jejunum preparations, compound 4 being the most active with an ED50 value of 0.09 +/- 0.001 mg/mL and 0.22 +/- 0.01 microg/mL on spontaneous and K+ -induced contractions, respectively. The cholinesterase inhibitory potential along with calcium antagonistic ability and safe profile in human neutrophil viability assay could make compounds 1-5 possible drug candidates for further study to treat Alzheimer's disease and associated problems.

Cytotoxic ergosterols from Paecilomyces sp. J300.

Seven ergosterol derivatives (1-7) were isolated from silkworm larvae infected with Paecilomyces sp. J300. On the basis of spectroscopic means, their structures have been elucidated as 3beta,5alpha-dihydroxy-ergosta-7,22-diene (1), 5alpha,6alpha-epoxy-(22E,24R)-ergosta-8(14), 22-diene-3beta, 7alpha-diol (2), 5alpha,6alpha-epoxy-(22E,24R)-ergosta-8,22-diene-3beta,7alpha-diol (3), ergosta-4,6,8(14),22-tetraene-3-one (4), ergosterol (5), ergosterol endoperoxide (6), 3beta,5alpha-dihydroxy-6beta-methoxyergosta-7,22-diene (7). Compounds 3-7 showed moderate cytotoxicity against five tumor cells.

Astasin, a novel cytotoxic carbohydrate-conjugated ergosterol from the colorless euglenoid, Astasia longa.

A novel cytotoxic carbohydrate-conjugated ergosterol (astasin) was found in cells of the colorless euglenoid, Astasia longa. Astasin accounted for about 2.4% of the total lipid of the cells. FAB-MS spectra of astasin showed MH+, 583.3387 (M+, C35H50O7). Astasin was composed of ergosterol (1 eq.), alpha-D-xylopyranose (1 eq.), and oxalic acid (1 eq.). By the acetylation using acetic anhydride and pyridine, oxalic acid was removed from astasin, and three hydroxyl groups of the xylopyranose moiety were acetylated. The two dimensional 13C- and 1H-NMR spectra suggest the oxalic acid was esterified with hydroxyl groups at C-2 and C-3 of the xylopyranose moiety and the hydroxyl group at C-1 of the xylopyranose was glycosidically linked to the hydroxyl group at C-3' of the ergosterol moiety. From the results, the structure of astasin was identified as 2,3-oxalyl-alpha-D-xylopyranosyl (1 --> 3')ergosterol. When cells of HL 60, a human lymphoma, were cultured with astasin, 50% of the cell growth was inhibited at 5.0 micrograms astasin/ml medium, and the cell growth was inhibited completely and 50% of the initial cells was killed at 10.0 micrograms astasin/ml medium.