Discovery of 23,24-diols containing ergosterols with anti-neuroinflammatory activity from Penicillium citrinum TJ507.

Citristerones A-E (1-5), five new 23,24-diols containing ergosterols, along with three known analogues, were isolated from the endophytic fungus Penicillium citrinum TJ507 obtained from Hypericum wilsonii N. Robson. Their structures and absolute configurations were determined by NMR, HRESIMS, Snatzke's method, X-ray diffraction analyses and ECD calculation. Subsequently, the anti-neuroinflammatory effects of these isolates were screened using lipopolysaccharide (LPS)-induced BV-2 microglial cells, and citristerone B (2) showed outstanding anti-neuroinflammatory activity, with IC50 value of 0.60 ± 0.04 µM. Moreover, immunofluorescence and western blot analysis suggested that citristerone B not only reduced the release of nitric oxide (NO) and proinflammatory cytokines in LPS-induced BV-2 microglial cells, but also significantly inhibited the expression of TNF-α, iNOS and NF-κB, along with the production of cellular ROS.

Two highly conjugated ergosterols from the fungus Psathyrella rogueiana and their anti-inflammatory activity.

Two previously undescribed ergosterols containing a highly conjugated ring system, psathrosterols A and B (1 and 2), have been isolated from the fungus Psathyrella rogueiana. Their structures with absolute configurations were established by extensive spectroscopic methods, as well as single crystal X-ray diffraction. Compounds 1 and 2 showed inhibitory activity against NO production with IC50 values of 22.3 and 16.4 µM, respectively.

Ergosterols from the Culture Broth of Marine Streptomyces anandii H41-59.

An actinomycete strain, H41-59, isolated from sea sediment in a mangrove district, was identified as Streptomyces anandii on the basis of 16S rDNA gene sequence analysis as well as the investigation of its morphological, physiological and biochemical characteristics. Three new ergosterols, ananstreps A-C (1-3), along with ten known ones (4-13), were isolated from the culture broth of this strain. The gross structures of these new compounds were elucidated on the basis of extensive analysis of spectroscopic data, including HR-ESI-MS, and NMR. The cytotoxicities of these isolates against human breast adenocarcinoma cell line MCF-7, human glioblastoma cell line SF-268, and human lung cancer cell line NCI-H460 and their antibacterial activities in inhibiting the growth of Candida albicans and some other pathogenic microorganisms were tested. Compounds 3-8, 10 and 11 displayed cytotoxicity with IC50 values in a range from 13.0 to 27.8 µg/mL. However, all the tested compounds showed no activity on C. albicans and other bacteria at the test concentration of 1 mg/mL with the paper disc diffusion method.

Ergosterols with rare peroxide, oxetane ring moiety, and a lactone ring from Aspergillus spectabilis and their immunosuppressive activities.

Ten ergostane-type steroids, including seven undescribed ones named spectasteroids A-G, were obtained from Aspergillus spectabilis. Their structures and absolute configurations were determined based on HRESIMS, NMR, ECD calculations, and single-crystal X-ray diffraction analyses. Structurally, spectasteroid A was a unique example of aromatic ergostane-type steroid that featured a rare peroxide ring moiety; spectasteroid B contained a rare oxetane ring system formed between C-9 and C-14; and spectasteroid C was an unusual 3,4-seco-ergostane steroid with an extra lactone ring between C-3 and C-9. Spectasteroids F and G specifically showed inhibitory effects against concanavalin A-induced T lymphocyte proliferation and lipopolysaccharide-induced B lymphocyte proliferation, with IC50 values ranging from 2.33 to 4.22 µM. Spectasteroid F also showed excellent antimultidrug resistance activity, which remarkable enhanced the inhibitory activity of PTX on the colony formation of SW620/Ad300 cells.

Psathrosterols C-E, highly conjugated ergosterol derivatives from Psathyrella rogueiana with anti-inflammatory and immunosuppressive activity.

Three new ergosterols featuring with a highly conjugated ring system, psathrosterols C-E (1-3), have been isolated from the fungus Psathyrella rogueiana. The structures with the absolute configurations were elucidated by means of spectroscopic methods and single crystal X-ray diffraction. Compounds 1-3 exhibit inhibitory activity against NO production with IC50 values ranging from 8.4 to 21.8 µM. Compound 1 inhibits the LPS-induced proliferation of B lymphocyte cells with an IC50 value of 12.3 µM.

Ten ring-B aromatized ergosterols from Aspergillus spectabilis.

Spectasterols F-O (1-10), ten interesting ergosterols with an aromatized B ring, were obtained from Aspergillus spectabilis. Their structures and absolute configurations were determined using a combination of high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), nuclear magnetic resonance (NMR) spectroscopy, single-crystal X-ray diffraction analyses, and electronic circular dichroism (ECD) calculations. Structurally, these aromatic ergosterols feature versatile side chains. Notably, compound aromatic ergosterols featured versatile side chains, and compound 4 is an unusual C23 ergosterol characterized by a shorter side chain due to oxidative cleavage between C-23 and C-24. All compounds were evaluated for their neuroprotective activities, with compound 8 showing a dose-dependent ability to reduce apoptosis and protect mitochondrial function in glutamate-induced SH-SY5Y cells.

Quadristerols A-G: Seven undescribed ergosterols from Aspergillus quadrilineata.

Quadristerols A-G, seven undescribed ergosterols, were obtained from Aspergillus quadrilineata. Their structures and absolute configurations were determined based on HRESIMS, NMR, quantum-chemical calculations, and single-crystal X-ray diffraction analyses. Quadristerols A-G featured ergosterol skeletons with different attachments; quadristerols A-C were three diastereoisomers possessing a 2-hydroxy-propionyloxy group at C-6, and quadristerols D-G were two pairs of epimers with a 2,3-butanediol group at C-6. All of these compounds were evaluated for their immunosuppressive activities in vitro. Quadristerols B and C showed excellent inhibitory effects against concanavalin A-induced T lymphocyte proliferation with IC50 values of 7.43 and 3.95 µM, respectively, and quadristerols D and E strongly inhibited lipopolysaccharide-induced B lymphocyte proliferation with IC50 values of 10.96 and 7.47 µM, respectively.

Anatoluin A and B isolated from medicinal Tricholoma anatolicum are new cytotoxic ergostanoids against the most common cancers.

Tricholoma anatolicum is an edible mushroom from the matsutake group growing under Cedar trees. Bioactivity-guided fractionation of Tricholoma anatolicum afforded two new (1 and 2), three known ergosterols (3-6), and four known (6-9) compounds. Structures were identified as anatoluin A (1), anatoluin B (2), 5α,6α-epoxy-ergosta-7,22-dien,3ß-ol (3), ergosterol-endoperoxide (4), ergosterol,3ß-ol (5), 3,5-dihydroxyfuran-2(5H)-one (6), mannitol (7), turanose (8), fumaric acid (9) using spectroscopic techniques. The cytotoxic activity of extract and isolated compounds was performed using MTT assay against MCF7, HT29, H1299, and HeLa cancerous cell lines while toxicity against PDF and L929 fibroblast healthy cell lines. The lipid peroxidation inhibitory and ABTS•+ scavenging activities were used to determine antioxidant activity. The polar extracts exhibited significant cytotoxic activity. The more perfect is that the extracts and isolated compounds (1-5) were inactive against PDF and L929 healthy cell lines. Compounds 1-3 and 4 exhibited noticeable cytotoxic activity, while 1-5 moderately inhibited lipid peroxidation.

Nine undescribed oxidized ergosterols from the endophytic fungus Penicillium herquei and their cytotoxic activity.

A chemical investigation of the EtOAc extract of the endophytic fungus Penicillium herquei led to the isolation of nine undescribed oxidized ergosterols, penicisterols A-I (1-9), along with ten known analogs (10-19). Their structures and absolute configurations were elucidated by a combination of spectroscopic data analysis, quantum-chemical electronic circular dichroism (ECD) calculations and comparisons, [Rh2(OCOCF3)4]-induced ECD experiments, DFT-calculated 13C chemical shifts and DP4+ probability analysis. Compound 1 was a rare example of ergosterol in which the bond between C-8 and C-9 was cleaved to form an enol ether. Moreover, compound 2 possessed a rare (2,5-dioxo-4-imidazolidinyl)-carbamic acid ester group substituted at C-3. All undescribed oxidized ergosterols (1-9) were evaluated for their cytotoxic activity against five cancer cell lines including 4T1 (mouse breast carcinoma), A549 (human pulmonary carcinoma), HCT-116 (human colorectal carcinoma), HeLa (human cervical carcinoma) and Hepg2 (human hepatoma carcinoma) cells. Compounds 2 and 3 displayed moderate cytotoxic activity against 4T1, A549 and HeLa cells, with IC50 values ranging from 17.22 to 31.35 µM.

New cytotoxic ergosterols from a plant-associated fungus Colletotrichum magnisporum.

Three new ergosterols, colletosterols A-C (1-3), together with two known analogues 4 and 5, were isolated from the endophytic fungus Colletotrichum magnisporum associated with the leaves of Rauvolfia verticillata by a bioassay-guided fractionation method. The new structures were elucidated on the basis of extensive spectroscopic analyses and electronic circular dichroism (ECD) calculations. All the ergosterols were evaluated for their cytotoxic activities against A549 and HeLa cell lines. Compounds 1-3 exhibited notable cytotoxicity with the IC50 values of 3.76-11.18 µM.

Chemical constituents and their biological activities from the mushroom Pyropolyporus fomentarius.

The chemical constituents and their biological activities of the mushroom Pyropolyporus fomentarius were investigated in this study. Two previously undescribed pentacyclic lupane-type triterpenes, 3-formyloxybetulin and 3-formyloxybetulinic acid, two rare degraded ergosterols, pyropolincisterols A and B, along with ten known triterpenoids and four known ergosterols were isolated from the fruiting bodies of P. fomentarius. Their chemical structures were determined using a combination of spectroscopic analysis. Nine compounds exhibited certain cytotoxicities to human cancer cell lines, while polyporenic acid showed significant cytotoxicities to SMMC-7721 and A-549 with IC50 values less than 10 µM. Four compounds showed inhibitory activities against nitric oxide (NO) production in LPS-activated RAW264.7 macrophages with IC50 values of 36.3, 25.1, 21.4, and 34.2 µM, respectively. The results of this assessment suggested that the lanostane triterpenoids and ergosterols in fruiting bodies of P. fomentarius played key roles in its folk usages.

Four new steroids from the leaves and twigs of Dysoxylum pallens and their cytotoxic activities.

Four previously undescribed steroids, identified as (3S,7S,8S,9S,10R,13S,14S,16S,17R,20S)-7α-methoxy-ergosta-5,24(28)-dien-3ß,16ß,20-triol (1), ergosta-5,24(28)-dien-3ß,7α,16ß-triol (2), ergosta-5,25-dien-3ß,7α,16ß,20-tetrol (3) and 7α,16ß,24α-trihydroxy-varninasterol (4), as well as five known analogues (5-9), were isolated from the leaves and twigs of Dysoxylum pallens Hiern (Meliaceae). Their structures were elucidated based on extensive spectroscopic analysis such as HR-ESI-MS, 1D and 2D NMR, UV, and IR. The absolute configuration of compound 1 was determined by X-ray diffraction analysis. Selected compounds were evaluated for their cytotoxic activities. Compounds 1, 2, and 8 exhibited moderate cytotoxic activity against HL-60, Hela, and HepG2 tumor cell lines with IC50 ranged from 11.09 to 17.51 µM.

Chemical constituents and their cytotoxicities from mushroom Tricholoma imbricatum.

Two undescribed triterpenes, tricholimbrins A and B, three undescribed steroids, tricholimbrins C‒E, one undescribed 4-chromanone derivative, along with 27 known compounds were isolated from fruiting bodies of the mushroom Tricholoma imbricatum. Tricholimbrins A and B are two polycyclic triterpenoids with a carbon degradation, while tricholimbrin C is a ring-rearranged steroid containing an aromatic moiety that might be derived from an ergosterol. Isocyathisterol, 3ß,15α-dihydroxyl-(22E,24R)-ergosta-5,8(14),22-trien-7-one, demethylincisterol A3, and volemolide showed cytotoxicities to six human cancer cell lines. 3ß-Hydroxyl-(22E,24R)-ergosta-5,8,22-trien-7,15-dione and 3ß-hydroxyl-(22E,24R)-ergosta-5,8,22-trien-7-one showed preferable cytotoxicities against HL-60 while chaxine C and volemolide showed preferable cytotoxicities against A-549, with IC50 values less than 10 µM.

Steroidal composition and cytotoxic activity from fruiting body of Cortinarius xiphidipus.

From the fruiting body of ectomycorrhizal fungi Cortinarius xiphidipus, sterols were identified from the crude extract and the cytotoxic effect of ergosta-4, 6, 8(14), 22-tetraen-3-one (ergone) was evaluated. Ten sterols including ergosta-3,5,7,9(11),22-pentaene, (22E)-ergosta-5,7,9(11),22-tetraen-3b-ol, (3ß,22E)-ergosta-5,7,22-trien-3-ol, (22E)-ergosta-7,22-dien-3-ol, neoergosterol, (3ß)-ergosta-5,8-dien-3-ol, (3ß)-ergosta-7-en-3-ol, stigmasterol, stigmasterol 22,23-dihydro and (22E)-ergosta-4,6,8(14),22-tetraen-3-one were identified from the crude extract. The cytotoxic activity of the sterol fraction containing ergosta-4, 6, 8(14), 22-tetraen-3-one was assessed on four tumour cell lines (Neuro-2a, Saos-2, MCF7 and LNCaP-C42). The cytotoxic activity against the four tumour cell lines tested, being Neuro-2a and Saos-2 the most sensitive, with a half-maximal inhibitory concentration (IC50) of 20.8 ± 2.2 and 27.8 ± 1.0 µg/mL, respectively. This is the first report of this Antarctic fungi collected in the Magallanes and Chilean Antarctica Region. This work represents a potential source for the development of anticancer drugs.

Anti-tumor and Anti-angiogenic Ergosterols from Ganoderma lucidum.

This study was carried out to isolate chemical constituents from the lipid enriched fraction of Ganoderma lucidum extract and to evaluate their anti-proliferative effect on tumor cells and human umbilical vein endothelial cells (HUVECs). Ergosterol derivatives (1-14) were isolated and purified from the lipid enriched fraction of G. lucidum. Their chemical structures were established by spectroscopic analyses or by comparison of mass and NMR spectral data with those reported previously. Amongst, compound 1 was purified and identified as a new one. All the compounds were evaluated for their anti-proliferative effect on human tumor cells and HUVECs in vitro. Compounds 9-13 displayed inhibitory activity against two types of human tumor cells and HUVECs, which indicated that these four compounds had both anti-tumor and anti-angiogenesis activities. Compound 2 had significant selective inhibition against two tumor cell lines, while 3 exhibited selective inhibition against HUVECs. The structure-activity relationships for inhibiting human HepG2 cells were revealed by 3D-QASR. Ergosterol content in different parts of the raw material and products of G. lucidum was quantified. This study provides a basis for further development and utilization of ergosterol derivatives as natural nutraceuticals and functional food ingredients, or as source of new potential antitumor or anti-angiogenesis chemotherapy agent.