Six New Polyhydroxysteroidal Glycosides, Anthenosides S1 - S6, from the Starfish Anthenea sibogae.

Six new polyhydroxysteroidal glycosides, anthenosides S1  - S6 (1 - 6), along with a mixture of two previously known related glycosides, 7 and 8, were isolated from the methanolic extract of the starfish Anthenea sibogae. The structures of 1 - 6 were established by NMR and HR-ESI-MS techniques as well as by chemical transformations. All new compounds have a 5α-cholest-8(14)-ene-3α,6ß,7ß,16α-tetrahydroxysteroidal nucleus and differ from majority of starfish glycosides in positions of carbohydrate moieties at C(7) and C(16) (1 - 4, 6) or only at C(16) (5). The 4-O-methyl-ß-d-glucopyranose residue (2) and Δ24 -cholestane side chain (3) have not been found earlier in the starfish steroidal glycosides. The mixture of 7 and 8 slightly inhibited the proliferation of human breast cancer T-47D cells and decreased the colony size in the colony formation assay.

Marine and semi-synthetic hydroxysteroids as new scaffolds for pregnane X receptor modulation.

In recent years many sterols with unusual structures and promising biological profiles have been identified from marine sources. Here we report the isolation of a series of 24-alkylated-hydroxysteroids from the soft coral Sinularia kavarattiensis, acting as pregnane X receptor (PXR) modulators. Starting from this scaffold a number of derivatives were prepared and evaluated for their ability to activate the PXR by assessing transactivation and quantifying gene expression. Our study reveals that ergost-5-en-3ß-ol (4) induces PXR transactivation in HepG2 cells and stimulates the expression of the PXR target gene CYP3A4. To shed light on the molecular basis of the interaction between these ligands and PXR, we investigated, through docking simulations, the binding mechanism of the most potent compound of the series, 4, to the PXR. Our findings provide useful functional and structural information to guide further investigations and drug design.

Tupisteroide A-C, three new polyhydroxylated steroidal constituents from the roots of Tupistra chinensis.

Three new steroidal compounds with polyhydroxy groups, tupisteroide A-C (1-3), were obtained from the roots of Tupistra chinensis, together with one known compound (4) that was isolated from this plant for the first time. The structures of tupisteroide A-C were determined on the basis of one- and two-dimensional NMR spectroscopy, including (1) H-(1) H Correlation Spectroscopy, Heteronuclear Multiple Bond Correlation, and Heteronuclear Single Quantum Coherence experiments. The isolated compounds were evaluated for their cytotoxic activities against A549, HepG2, and CaSki cancer cell lines in vitro. Among them, compounds 1, 2, and 4 did not show significant inhibitory activity, but compound 3 showed cytotoxicity against A549 cancer cell lines with IC(50) values of 25.0 µM.

New 3beta,5alpha,6beta-trihydroxysteroids from the octocorals Bebryce sp. (Plexauridae) and Carijoa sp. (Clavulariidae).

A new 3beta,5alpha,6beta-trihydroxysteroid, bebryceoid A (1), has been isolated from an octocoral Bebryce sp. In addition, an octocoral Carijoa sp. yielded two new 3beta,5alpha,6beta-trihydroxysteroids, carijoids A (2) and B (3). The structures of steroids 1-3 were elucidated by spectroscopic methods and by comparison of the spectral data with those of known steroid analogues.

Bendigoles A approximately C, new steroids from Gordonia australis Acta 2299.

Bendigoles A approximately C are the first secondary metabolites to be isolated from a member of the actinomycete genus Gordonia. They were detected in a culture filtrate extract of Gordonia australis Acta 2299 by HPLC-diode array analysis and characterized as new steroids by mass spectrometry and NMR experiments. Bendigole C show binding affinity to the human progesterone and A approximately C to androgen receptor but are inactive at mineralocorticoid and estrogen receptors. In in vitro transactivation studies bendigoles A and C showed moderate and weak androgenic activities.

Sinugrandisterols A-D, trihydroxysteroids from the soft coral Sinularia grandilobata.

Four new trihydroxysteroids, sinugrandisterols A-D (1-4), have been isolated from the CH(2)Cl(2)-soluble fraction of the EtOH extract of Sinularia grandilobata. The structures of these metabolites were determined on the basis of spectroscopic (IR, MS, and 1D and 2D NMR) analysis. The cytotoxicity of 1-4 toward a limited panel of cancer cell lines is also reported.

Influence of polyhydroxysteroids on [Ca(2+)](i).

Recently, we have shown that two biologically active, disulfated polyhydroxysteroids from the Pacific brittle star Ophiopholis aculeata stimulate Ca(2+) influx into different cell types. In the present study, 45Ca(2+) and two fluorescent calcium probes, quin-2/AM and fura-2/AM, were employed to investigate the course and amplitude of calcium signals induced in different mouse cells using an radio-isotope, spectrofluorimetry, and microcytofluorimetry techniques. The cytotoxic and hemolytic effects were not observed for both steroids at the wide range of concentrations. Steroids did not influence [3H]-uridine incorporation in a variety of cells. The investigated steroids stimulated a rapid increase in cytosolic Ca(2+) content in Ehrlich mouse carcinoma cells, mouse spleen lymphocytes, and mouse peritoneal macrophages in the concentration range 1-100 microg/ml on a dose-dependent basis. Blockers of L-type calcium channels, such as verapamil, diltiazem, nifedipine (1 x 10(-7)M), and 1mM EGTA, inhibited this process and reduced the response of cells to steroid application. The stimulatory effect of steroids on human fibroblast proliferation and mouse macrophage lysosome activity was observed also. It is suggested that the investigated compounds may act as Ca(2+)-agonists and increase Ca(2+)-transport across cell membranes.

Evidence for 16beta-hydroxylation of estrogens by guinea pig liver slices.

In an earlier communication, we conclusively proved that estrone-3-sulfate could be hydroxylated in the 16alpha position by guinea pig liver slices. Disulfates of 16alpha-hydroxyestrone and estriol were identified. Suggestive evidence for 6-hydroxylated metabolites was also mentioned. A careful reinvestigation has now shown that the steroid disulfate fraction is composed of 16alpha- and 16beta-hydroxylated steroids. 16beta-Hydroxyestrone in particular is an important quantitative metabolite of estradiol-17 beta. It is concluded that no firm evidence is available for the formation of 6-hydroxysteroids in the tissue system under consideration.

Application of over-run thin-layer and gas-liquid chromatography in the separation of closely related C19O2 steroids.

An improved method for the separation of epimeric C19O2 steroids and their related allylic alcohols is described. In this method, the steroids are first separated by over-run thin-layer chromatography, and the unresolved groups are further analysed as free or as trimethylsilyl ether derivatives by gas-liquid chromatography. The behaviour of twenty-one C19O2 steroids was investigated by thin-layer chromatography in four systems and by gas-liquid chromatography in four liquid phases. All steroid pairs of similar polarity were resolved by the combination of these two fractionation procedures.

Estrogen biosynthesis from testosterone-4-14C by post-ovulatory human ovaries. Comparison between the corpus-luteum ovary and its partner.

PIP: The whole of each ovary from 2 women in the luteal phase of normal cycles was incubated with testosterone-4-carbon-14 in order to test the postulate that an interaction between ovarian compartments might be involved in the post-ovulatory synthesis of estrogens. Estradiol, estriol and estrone were all identified, estriol in only one ovary, from the corpus-luteum ovary. In both women, the corpus-luteum ovary converted more than 60 times as much substrate to estrogens than its non-corpus-luteum partner, and produced markedly lower yields of testosterone, 4-androstene-3, 17-dione(androstene-dione) and of their 16-alpha-hydroxy derivatives. All 4 of these compounds were identified. The 19-hydroxy derivative of testosterone and androstene-dione were also identified, but with a less marked difference between the two ovaries. The markedly lower yields of the 16-alpha-hydroxy derivatives from the corpus-luteum ovaries indicates that they were being further metabolized. The yield of estriol was too low, however, to implicate this steroid as a final product of their conversion.^ieng