Zhangshuang Deng

Estrogenic and anti-estrogenic activities of hispolon from Phellinus lonicerinus (Bond.) Bond. et sing.

Hispolon was the main antitumor active ingredient in Phellinus sensu lato species. In order to confirm the dual regulating estrogenic ingredient and obtain more effective natural estrogen replacement drugs, hispolon was separated from Phellinus lonicerinus (Bond.) Bond. et sing. Hispolon exhibited significant anti-proliferative effect against estrogen-sensitive ER (+) MCF-7 cells in the absence of estrogen, and exhibits antagonistic effects on 17β-estradiol (E2)-induced MCF-7 cell proliferation when E2 and the different concentrations of hispolon were treated simultaneously. Hispolon also inhibited the proliferation of estrogen-negative ER (-) MDA-MB-231 cells at the concentration of 5.00×10(-5) M. The yeast two-hybrid experiments showed that hispolon had strong and non-selective effects on the estrogen receptor (ER) α and ERβ at a concentration of 1.00×10(-6) M. The ERβ-binding ability of hispolon was larger than ERα in the concentration range of 1.00×10(-9) M and 1.00×10(-7) M. Hispolon could increase the body weight coefficient, serum E2 and progesterone contents in immature female mice at dose of 9.10×10(-6) mol/kg, and increase coefficient of thymus and spleen in mice. The Gscores of hispolon-ERα and hispolon-ERβ docked complexes were -7.93 kcal/mol and -7.79 kcal/mol in docking simulations. Hispolon presented dual regulating estrogenic activities, which showed estrogenic agonist activity at low concentration or lack of endogenous estrogen, and the estrogenic antagonistic effect was stimulated at high concentrations or too much endogenous estrogen. Hispolon could be used for treating the estrogen deficiency-related disease with the benefit of non-toxic to normal cells, good antitumor effects and estrogenic activity.

Paraconfuranones A-H, eight new furanone analogs from the insect-associated fungus Paraconiothyrium brasiliense MZ-1.

Insects that constituted the largest number of organisms on earth possess the richest and most unexplored domains of natural products.Microorganisms livingwithin the insect inner gut system without causing diseases have proven to be potential sources of diverse chemical structures, for example, unprecedented immunosuppressive polyketides from a mantis-associated fungus Daldinia eschscholzii, antifungal naphthalenones from Perenniporia sp. inhabiting the larva of Euops chinesis, cytotoxic angucyclines from Amycolatopsis sp.HCa1, anda rare actinobacteriumderived fromthe gutofOxyachinensis. Asapartofourongoingpiecesof researchon the utilization of fungal sources in Shennongjia Forest District, a National Natural Protection Region of China, a rare fungus Paraconiothyrium brasiliense (strain no. MZ-1) was isolated from the gut of healthy Acrida cinerea collected from the Shennongjia Forest District. To our best knowledge, the strain can produce some novel terpenoid derivatives. When this fungus was cultured under the rice medium in our preliminary experiments, a few terpenoids could be detected. When the culture medium was guided by the one strain–many compounds approach, the secondary metabolites thereby changed. Further investigation led to the isolation of eight new furanone derivatives, named paraconfuranones A–H (Fig. 1). In thispaper,we reported the isolationandstructural elucidation of paraconfuranones A–H and their cytotoxity activity.

Machilusmarin, a new neuroprotective isocoumarin dimer from the stems of Machilus ichangensis Rehd. et Wils.

A new isocoumarin dimer, Machilusmarin (1), was isolated from the stem of Machilus ichangensis Rehd. et Wils., together with two known coumarins, scopoletin (2) and isoscopoletin (3). The structure of Machilusmarin (1) was elucidated by HRMS and NMR spectrums. Machilusmarin (1) showed significant neuroprotective activity against SH-SY5Y human neuroblastoma cells apoptosis induced by hydrogen peroxide at concentrations ranging from 0.625 to 10.0u2009µgu2009mL−1.

Cytotoxic tremulanes and 5,6-secotremulanes, four new sesquiterpenoids from a plant-associated fungus X1-2.

Abstract Two new tremulanes and two new 5,6-secotremulanes, davotremulanes A-D 1–4, along with four known compounds 5–8, were isolated from the culture extract of X1-2, an unidentified plant-associated fungus, which was isolated from the endangered plant, Davidia involucrate Baill. in Shennongjia District. The structures of new compounds 1–4 were established on the basis of extensive spectroscopic analysis. Compounds 1–8 were evaluated for cytotoxic activity to four cancer cell lines, and compounds 1, 2 and 5 displayed selectively moderate activities to A549 cell line with IC50 at 15.3, 25.2, 35.2 μg/mL.

A new cinnamic acid derivative from plant-derived endophytic fungus Pyronema sp.

Abstract Ten secondary metabolites (1–10) including a new 4-hydroxycinnamic acid derivatives, methyl 2-{(E)-2-[4-(formyloxy)phenyl]ethenyl}-4-methyl-3-oxopentanoate (1), and nine known compounds (2–10) were isolated from an EtOAc extract derived from a solid rice medium of endophytic fungal strain Pyronema sp. (A2-1 & D1-2). Their structures were elucidated from NMR and HRMS data. All the compounds were tested for antibacterial activity against Mycobacterium marinum ATCCBAA-535. Compounds 1, 8 and 9 exhibited moderate antibiotic activity with IC50 of 64, 43 and 32 μM, respectively.

Structural elucidation and NMR spectral assignments of paraconfuranones I-M from the insect-associated fungus Paraconiothyrium brasiliense.

Furanones consisted of natural oxygenated heterocycles have been reported from various natural sources. Fungus-derived furanone metabolites include aspertetronins A-B and gregatins A-E isolated from Cephalosporium gregatum, graminin A from Cephalosporium gramineum, 704-I-704-IV from Aspergillus panamensis, penicilliols A-B from penicillium daleae K. M. Zalessky, huaspenones A-B from Aspergillus sp. XW-12, and graminin B from Paraconiothyrium sp. Microorganisms living within the insect inner gut system without causing diseases have proven to be potential sources of diverse chemical structures. In the course of our systematic researches on the utilization of Chinese herbs and fungal sources in Shennongjia Forest District, a National Natural Protection Region of China, a rare fungus Paraconiothyrium brasiliense was isolated from the gut of healthy Acrida cinerea collected from the aforementioned district. It should be noted that the genus Paraconiothyrium have rarely been chemically explored, some terpenoids and furanones had been reported to date. In the previous paper, we reported the isolation and structural elucidation of paraconfuranones A–H from the insectassociated fungus P. brasiliense. During the continual screening for bioactive furanone analogs, another five new furanones, paraconfuranones I–M (3–7), together with two known furanones (1–2) were obtained from the genus under different fermentation conditions. Furthermore, the neuroprotective activities of the isolated compounds 1–3 and 5–6 were evaluated by PC12 cell apoptosis induced by hydrogen peroxide, and the compound 3 exhibited significant neuroprotective activities. In this paper, we reported the isolation and complete assignments of H and C-NMR spectral data of compounds 1–7 by MS, 1D-NMR and 2D-NMR spectral analyses, and the neuroprotective activity evaluation of these compounds against hydrogen peroxideinduced PC12 cell damage.

A New Wutaifuranol Derivative From Solanum cathayanum

The plant Solanum cathayanum is widely distributed in China, and the whole plant has been clinically available in Chinese folk medicine as anti-cancer, anti-inflammatory, and anti-bacterial agents. Our previous phytochemical investigations of the plant revealed the existence of phenylpropanoids, coumarins, lignans, and alkaloids [1]. Recently, further exploitation of the chemical constitutions of Solanum cathayanum led to the discovery of 3-hydroxymethyl-7-methoxywutaifuranol (1), a new wutaifuranol analogue. Wutaifuranol and its derivatives are characteristic members of the benzofuran family and are widespread in nature. They exhibit a broad range of biological activities such as anticancer [2], anti-inflammatory [3], antifungal [4], and antitubercular [5]. Herein, we report the detailed isolation and structural elucidation of 1. The dried Solanum cathayanum (20 kg) was extracted with 95% ethanol under reflux for three times. After removal of solvent under reduced pressure, the extract was suspended in water and degreased with ligarine. The water layer was acidified with 4% HCl and filtered. The alkalized filtrate, pH adjusted to 7.0 with Na2CO3, was extracted with chloroform for three times. The combined organic layers were concentrated to give a residue (400 g). The crude residue (20 g) was chromatographed on a reverse-phase column eluting with 5–70% CH3CN–H2O to give 153 fractions. Fractions 26–28 were combined and further separated by semipreparative reverse-phase HPLC on a Diamonsil C18 column (5 m, 250 4.6 mm) eluting with 17.5% CH3CN–H2O (v/v) to afford compound 1 (5 mg). Compound 1 was isolated as a yellow amorphous powder, and its molecular formula was established as C13H14O4 by HR-ESI-MS (m/z [M + H]+ 235.0960, calcd 235.0970) and NMR spectrum. The 1H NMR spectrum revealed two meta-coupled protons characteristic of a benzene ring at 7.25 (1H, d, J = 1.2 Hz, H-4) and 7.02 (1H, d, J = 1.2 Hz, H-6), a trans-3-hydroxy-1-propenyl group [ 6.60 (1H, d, J = 15.8 Hz, H-8), 6.37 (1H, dt, J = 15.8 and 5.2 Hz, H-9) and 4.14 (2H, m, H-10)], an alkene proton at 7.79 (1H, s, H-2), an oxygenated methylene proton at 4.59 (2H, d, J = 3.5 Hz, H-11) in the downfield region, a methoxy proton at 3.95 (3H, s, 7-OCH3), and two exchangeable proton at 4.82 (1H, br.s, 10-OH) and 5.11 (1H, br.s, 11-OH). The 13C NMR and DEPT spectra displayed five olefinic methine carbons at 142.7, 110.5, 104.8, 129.7, and 129.0, two oxygenated methylene carbons at C 61.5 and 53.8, a methoxy carbon at 55.8, and five sp2 quaternary carbon at 144.9, 143.5, 132.8, 128.7, and 121.9. The 1H and 13C NMR spectra of compound 1 (Table 1) were similar to those of 7-methoxywutaifuranol [5], having a benzofuran framework, except that a proton of furan ring was observed at 7.79 (1H, s, H-2). Inspection of the HMBC spectrum revealed that a hydroxymethyl group signal at H 4.59 (2H, d, J = 3.5 Hz, H-11) and 5.11 (1H, br.s, 11-OH) in 1 replaced the ortho-coupled protons in the furan ring of the benzofuran unit in 7-methoxywutaifuranol. The location of the hydroxymethyl group at C-3 in the furan ring was determined by the HMBC correlations (Fig. 1) from H-11 ( 4.59) to C-3 ( 121.9), C-3a ( 128.7), and C-2 ( 142.7). On the basis of the above evidence, the structure of compound 1 was elucidated as the hitherto unreported 3-hydroxymethyl-7-methoxywutaifuranol.

Unusual C25 Steroids with Bicyclo[4.4.1]Skeleton at A/B Rings from Endophytic Fungus Penicillium citrinum

Endophytic fungus isolated from medicinal plants are a fertile source of natural products with novel structures and interesting biological activities [1]. Recently, 14 endophytic filamentous fungi were isolated from tissue fragments of the Chinese medicinal plant species Saccharum arundinaceum Retz. collected from Yichang, Hubei Province in April 2011 and identified based on morphological traits and internal transcribed spacer rDNA-ITS sequence analysis. The EtOAc extracts of all fungal cultures were evaluated for cytotoxictiy using the MTT method and an extract of Penicillium citrinum BM7 exhibited cytotoxic activity with 85% inhibition of HepG2 cell and 65% inhibition of CaSki cell at 100 g/mL. HPLC-DAD-guided investigation of secondary metabolites of the fungus Penicillium citrinum BM7 led to the discovery of a series of unusual C25 steroids with bicyclo[4.4.1]skeleton at A/B rings. Herein, we present details on the isolation and structural identification of all seven steroids (1–7). The EtOAc extract prepared from the solid fermentation products of the endophytic fungus Penicillium citrinum BM7 was subjected repeatedly to column chromatography on silica gel, Sephadex LH-20, and preparative HPLC to afford compounds 1–7, which were identified as erythro-23-O-methylneocyclocitrinol (1), threo-23-O-methylneocyclocitrinol (2), 22-acetylisocyclocitrinol A (3), isocyclocitrinol A (4), isocyclocitrinol B (5), neocyclocitrinol A (6), and neocyclocitrinol B (7) by extensive analysis of their spectroscopic data, and all these data were in good agreement with literature data. erythro-23-O-Methylneocyclocitrinol (1). Colorless powder, C26H38O4. ESI-MS m/z 415 [M + H]+. 13C NMR (100 MHz, CDCl3, , ppm): 205.0 (C-6), 157.35/157.30 (C-8), 145.8 (C-10), 141.11/141.06 (C-20), 124.9 (C-7), 123.37/123.03 (C-22), 122.1 (C-1), 81.21/81.17 (C-23), 69.42/69.36 (C-24), 64.6 (C-3), 59.7 (C-17), 56.17/56.12 (-OCH3), 55.4 (C-14), 54.1 (C-9), 48.6 (C-5), 47.07/47.04 (C-13), 41.7 (C-4), 38.00/37.91 (C-12), 35.8 (C-2), 27.76 (C-18), 27.67 (C-11), 24.39/24.31 (C-16), 22.8 (C-15), 18.66/18.17 (C-21), 17.82/17.77 (C-25), 13.8 (C-19) [2].

Secondary Metabolites from the Endophytic Fungus Penicillium citrinum

Endophytic fungi represent a virtually inexhaustible and sustainable resource of bioactive and chemically novel natural products with potential for utilization in the medical field [1]. Our systemic investigation of secondary metabolites from this fungi led to the discovery of (3R,4S)-6,8-dihydroxy-3,4,5,7-tetramethylisochroman (1), decarboxydihydrocitrinin (2), alternethanoxin A (3), 2,3,4-trimethyl-5,7-dihydroxy-2,3-dihydrobenzofuran (4), penicitrinone A (5), penicitrinol A (6), citrinolactone B (7), -diversonolic ester (8), emodin (9), citreorosein (10), 2,3,5-trimethyl-6-(3-oxobutan-2-yl)-4H-pyran-4one (11), kojic acid (12), and phenol A (13). Herein, we report details on the isolation and structural elucidation of these compounds. The fungus Penicillium citrinum BM7 (strain No. JX966317) was isolated from inner tissue fragments of the plant species Saccharum arundinaceum Retz. collected in Yichang, Hubei Province in April 2011 and identified based on morphological traits and internal transcribed spacer rDNA-ITS sequence analysis. The fungus was grown on solid rice medium at room temperature under static conditions for 30 days, and the metabolites were extracted with EtOAc. The crude extracts (60 g) were repeatedly chromatographed on silica gel columns and extensive reversed-phase semipreparative HPLC to afford the title compounds. The structures of these compounds were identified by extensive analysis of their spectroscopic data, which were in good agreement with literature data. Alternethanoxin A (3) and 2,3,5-trimethyl-6-(3-oxobutan-2-yl)-4H-pyran-4-one (11) have been isolated from the Penicillium citrinum for the first time. (3R,4S)-6,8-Dihydroxy-3,4,5,7-tetramethylisochroman (1). Yellow powder, C13H18O3. ESI-MS m/z 245 [M + Na]+ [2]. Decarboxydihydrocitrinin (2). Yellow powder, C12H16O3. ESI-MS m/z 209 [M + H]+ [3]. Alternethanoxin A (3). Yellow powder, C16H14O6. ESI-MS m/z 303 [M + H] +. 1H NMR (400 MHz, CDCl3, , ppm, J/Hz): 9.09 (1H, s, 6-OH), 7.50 (1H, dd, J = 7.7, 1.0, H-8), 7.35 (1H, dd, J = 8.0, 7.7, H-9), 7.13 (1H, dd, J = 8.0, 1.0, H-10), 6.23 (2H, s, H-3, 6), 3.65 (3H, s, 7-OCH3), 2.24 (3H, s, COCH3). 13C NMR (100 MHz, CDCl3, , ppm): 198.9 (COCH3), 167.0 (C-7), 160.2 (C-4, 5), 153.6 (C-6a), 148.8 (C-1), 130.9 (C-9), 130.3 (C-10a), 129.0 (C-2), 122.1 (C-8), 121.4 (C-10), 109.7 (C-11), 109.4 (C-3, 6), 52.7 (7-COH3), 22.1 (COCH3) [4]. 2,3,4-Trimethyl-5,7-dihydroxy-2,3-dihydrobenzofuran (4). Yellow powder, C11H14O3. ESI-MS m/z 195 [M + H] + [5]. Penicitrinone A (5). Yellow powder, C23H24O5. ESI-MS m/z 381 [M + H]+ [3]. Penicitrinol A (6). Yellow powder, C23H26O5. ESI-MS m/z 383 [M + H]+ [3].

Estrogenic and Antiestrogenic Activities of Protocatechic Acid

Study the estrogenic and antiestrogenic effects of protocatechuic acid with the aim of obtaining a safe and effective natural estrogen replacement drugs. Its estrogenic and antiestrogenic effects were evaluated through cell proliferation experiments. The estrogen-receptor (ER) binding abilities of protocatechuic acid were tested by yeast two-hybrid experiment, and their possible binding sites for ERs were performed by computer-aided molecular docking technology. Protocatechuic acid showed significant effects on the proliferation of estrogen-sensitive ER (+) MCF-7 cells in the absence of estrogen, and resulted in antagonistic effects on E2-induced MCF-7 cell proliferation. However, it could not induce the proliferation of estrogen-negative ER (-) MDA-MB-231 cells. The yeast two-hybrid experiments showed protocatechuic acid had significant but non-selective binding abilities for the two ERs. Protocatechuic acid revealed a double directional adjusting function of estrogenic and antiestrogenic activities, which showed estrogenic agonist activity at low concentration or lack of endogenous estrogen, and the estrogenic antagonistic effect was stimulated at high concentrations or too much endogenous estrogen. Protocatechuic acid had significant binding capacity for ERs. Therefore, protocatechuic acid could be used in the treatment of the estrogen deficiency-related diseases.