Steroidal monoglycosides from the Far Eastern starfish Hippasteria kurilensis and hypothetic pathways of polyhydroxysteroid biosynthesis in starfish

Five new steroidal monoglycosides, kurilensosides E (1), F (2), G (3), H (4) and 15-O-sulfate of echinasteroside C (5) were isolated along with the previously known echinasteroside C (6) from the alcoholic extract of the Far Eastern starfish Hippasteria kurilensis collected near Kuril Islands. Compounds 1-3 were determined to contain unusual polyhydroxysteroidal aglycons lacking 6-hydroxy group. Aglycon moiety of kurilensoside H (4) was shown to be the first case of marine polar steroids containing 4,5-epoxy functionality. Hypothetic pathways of the biosynthesis of polyhydroxysteroids and related glycosides in starfish and the existence of the late C-6 oxidation pathway in H. kurilensis are discussed.     

Anti-inflammatory components of the Vietnamese starfish Protoreaster nodosus

Background

In the present study, we examined the inhibitory effects of a methanolic extract, dichloromethane fraction, water layer, and polyhydroxylated sterols (1–4) isolated from the Vietnamese starfish Protoreaster nodosus on pro-inflammatory cytokine (IL-12 p40, IL-6, and TNF-α) production in LPS-stimulated bone marrow-derived dendritic cells (BMDCs) using enzyme-linked immunosorbent assays (ELISA).

Results

The methanolic extract and dichloromethane fraction exerted potent inhibitory effects on the production of all three pro-inflammatory cytokines, with IC₅₀ values ranging from 0.60 ± 0.01 to 26.19 ± 0.64 μg/mL. Four highly pure steroid derivatives (1–4) were isolated from the dichloromethane fraction and water layer of P. nodosus. Potent inhibitory activities were also observed for (25S) 5α-cholestane-3β,4β,6α,7α,8β,15α,16β,26-octol (3) on the production of IL-12 p40 and IL-6 (IC_50s = 3.11 ± 0.08 and 1.35 ± 0.03 μM), and for (25S) 5α-cholestane-3β,6α,8β,15α,16β,26-hexol (1) and (25S) 5α-cholestane-3β,6α,7α,8β,15α,16β,26-heptol (2) on the production of IL-12 p40 (IC_50s = 0.01 ± 0.00 and 1.02 ± 0.01 μM). Moreover, nodososide (4) exhibited moderate inhibitory effects on IL-12 p40 and IL-6 production.

Conclusion

This is the first report of the anti-inflammatory activity from the starfish P. nodosus. The main finding of this study is the identification oxygenated steroid derivatives from P. nodosus with potent anti-inflammatory activities that may be developed as therapeutic agents for inflammatory diseases.     

Bioactive polyhydroxylated sterols from the marine sponge Haliclona crassiloba

Four new polyhydroxylated sterols, named halicrasterols A–D (1–4), together with six known analogs (5–10) were isolated from the marine sponge Haliclona crassiloba. Compounds 1 and 2 represented rare examples of steroids featuring 17(20)E-double bonds. The structures of 1–10 were elucidated by spectroscopic analysis and comparison with reported data. This is the first report of a steroid profile for this species. The antimicrobial activities of 1–10 were evaluated against a panel of bacterial and fungal strains in vitro, and compounds 4 and 9 showed moderate activity against some of the Gram-positive strains with MICs ranging from 4 to 32 μg/mL.     

Polyhydroxylated steroids from the soft coral Sinularia dissecta

A repeated silica gel column chromatography followed by HPLC purification on the methanol extract of marine soft coral Sinularia dissecta, resulted in the isolation of fifteen polyhydroxylated steroids (1-15), involving six new C-18 functionalized steroids, 3beta-acetoxy-1alpha,11alpha-dihydroxygorgost-5-en-18-oic acid (1), gorgost-5-en-1alpha,3beta,11alpha,18-tetrol (2), 18-acetoxy-1alpha,3beta,11alpha-trihydroxygorgost-5-ene (3), 24(S)-3beta-acetoxy-1alpha, 11alpha-dihydroxyergost-5-en-18-oic acid (4), 24(S)-ergost-5-en-1alpha,3beta,11alpha,18-tetrol (5), and dissectolide (6). The structures of the new compounds were determined on the basis of extensive spectroscopic data (IR, MS, (1)H and (13)C NMR, HMQC, HMBC, and NOESY) analysis. Compound 6 was found as an unusual sterol bearing a lactone functionality.     

New 19-oxygenated and 4-methylated steroids from the Formosan soft coral Nephthea chabroli

Chemical investigation of the Formosan soft coral Nephthea chabroli resulted in the isolation of four new 19-oxygenated steroids, nebrosteroids I-L (1-4), together with a new 4α-methylated steroid, nebrosteroid M (5). The molecular structures of these isolated metabolites were elucidated on the basis of extensive spectroscopic analysis and by comparison of the data with those of related metabolites. Compounds 1-5 were evaluated for anti-inflammatory activity using RAW 264.7 macrophages.     

Seasonality of reproduction in wild boar (Sus scrofa) assessed by fecal and plasmatic steroids

The collection of biological samples through non-invasive techniques represents one way of monitoring in vivo physiological changes associated with reproductive activity. Such techniques are particularly important for the study of animal species in the wild.The goals of this study were 1) to evaluate fecal progestogen (P), estrogen (E), and androgen (A) by means of radioimmunoassays, in male and female wild boars culled in the Piedmont, Italy area; 2) to compare them with plasmatic concentrations and the animals’ reproductive status; and 3) to assess variations in reproductive seasonality between two populations of wild boars living in a mountainous vs. a plain habitat in Piedmont.The results demonstrate a positive correlation between fecal and plasmatic steroid concentrations (r = 0.46, 0.58, and 0.45 for plasma P₄ and P, E₂ and E, and T and A; P < 0.05). Moreover, high fecal levels of both P and E (>170 ng/g and >100 pg/g respectively) were found in 70.6% of pregnant sows and in none of the non-pregnant animals, thus supporting the use of this technique for detecting pregnancy status in wild boar.Similar birth patterns were displayed by the mountain and plain populations, but births peaked significantly only in the mountain population, in the spring (46%, P < 0.05, vs. other seasons). A corresponding autumnal peak of plasma testosterone concentrations in males was displayed only by the mountain population (7.4 vs. < 2.0 ng/mL in the other seasons, P < 0.05).The correlation between fecal and plasmatic steroid concentrations obtained in this study supports the applicability of this non-invasive sampling technique for monitoring reproductive status in wild boar, thus enabling a more informed and correct management of the species.     

3-Keto steroids from the marine organisms Dendrophyllia cornigera and Cymodocea nodosa

The new (20R)-22E-cholesta-4,22-diene-3,6-dione (1), along with three known 3-keto steroids were isolated from the deep-water Mediterranean scleractinian coral Dendrophyllia cornigera (2-4). Moreover, four known related 3-keto steroids were isolated from the sea grass Cymodocea nodosa (5-8). The structure elucidation of steroid 1 and the full NMR resonance assignments of all isolated metabolites were based on interpretation of their spectral data. All compounds are reported for the first time as metabolites of the investigated organisms. Compounds 2 and 3 showed significant cytotoxicity against lung cancer NSCLC-N6 cell line.     

New 4alpha-methyl steroids from a Chinese soft coral Nephthea sp

Three new 4alpha-methyl steroids, 4alpha-methyl-ergost-7,24(28)-diene-3beta-ol-23-one (1), 4alpha-methyl-ergost-8(14),24(28)-diene-3beta-ol-23-one (2), 4alpha-methyl-ergost-24(28)-en-3beta,11beta-diol-23-one (3), and two new polyoxygenated steroids, ergost-5,25-diene-3beta,24S,28-triol (4), ergost-5,24(28)-diene-3beta,23S-diol (5), along with one known steroid 24-methylencholesterol (6) were isolated from the soft coral Nephthea sp. Their structures including the relative configurations were elucidated by extensive 1D and 2D NMR spectra analysis. The absolute configurations of (4) and (5) were determined by the Mosher's method.     

New pregnane steroids from Turraea pubescens

Three new pregnane steroids, 2beta,3beta,5beta-trihydroxy-pregn-20-en-6-one (1), 3beta-hydroxy-5alpha-pregn-7,20-dien-6-one (2), and 3beta-acetoxy-5alpha-pregn-7,20-dien-6-one (3) were isolated from the twigs and leaves of Turraea pubescens, and were structurally elucidated on the basis of spectroscopic data and chemical method.     

Polyhydroxylated steroids from an endophytic fungus, Chaetomium globosum ZY-22 isolated from Ginkgo biloba

A novel polyhydroxylated C₂₉-sterol, 25ξ-methyl-22-homo-5α-cholest-7,22-diene-3β,6β,9α-triol, designated globosterol (1), together with one known tetrahydroxylated ergosterol (22E, 24R)-ergosta-7,22-diene-3β,5α,6β,9α-tetraol (2) has been isolated from the cultures of an endophytic fungus, Chaetomium globosum ZY-22 originated from the plant Ginkgo biloba. The structures and relative configurations of 1 and 2 were established on the basis of extensive spectroscopic analyses including 1D and 2D NMR (¹H-¹H COSY, HSQC, HMBC, and NOESY) experiments and comparison with the literature. Globosterol (1) possesses an unprecedented 25-methyl Δ²²-C₁₀-side chain and Δ⁷-3β,6β,9α-hydroxy-steroid nucleus, which represents the first example for C₂₉-steroids of the group.     

New bufadienolides and C23 steroids from the venom of Bufo bufo gargarizans

Six new bufadienolides (1-6) and two new C₂₃ steroids (7 and 8), together with three known compounds (9-11) were isolated from the venom of Bufo bufo gargarizans. Their structures were elucidated by spectroscopic analysis and single-crystal X-ray diffraction. In vitro cytotoxicities of all compounds were evaluated in A549 cancer cell line. Compounds 2, 3 and 10 showed significant cytotoxic activities.     

Carrageenan from Sarconema scinaioides (Gigartinales, Rhodophyta) of Indian waters

Carrageenan was extracted from the red seaweed Sarconema scinaioides of Indian waters and was characterized. The crude carrageenan as well as its alkali modified derivative was composed of 3,6-anhydro galactose, 6-O-methyl galactose as well as galactose moieties in various proportions. Linkage analysis exhibited that these two carrageenan samples consisted of 4-linked 3,6-anhydrogalactose residue sulphated at position 2, and 3-linked galactose residue sulphated at position 4. The physicochemical and rheological data along with molecular weight data, FT-IR, 1D and 2D NMR (¹H, ¹³C, COSY and HSQC) spectrometry suggested that the polysaccharide was composed predominantly of iota- along with a small amount of its precursor nu (ν)-carrageenan, unlike the hybrid carrageenans (iota-, pyruvated- and kappa-carrageenans) from this seaweed reported in the literature. This Indian seaweed species would be a potentially important source of iota-carrageenan.     

Cariniferosides A–F and other steroidal biglycosides from the starfish Asteropsis carinifera

Six new steroidal biglycosides, cariniferosides A–F (1–6), were isolated along with six previously known glycosides, 7–12, from the alcoholic extract of the starfish Asteropsis carinifera. The structures of 1–6 were determined by extensive NMR and ESIMS techniques. The isolated compounds did not show any apparent cytotoxicity in cancer cell lines HCT-116, RPMI-7951, and T-47D, but sulfated compounds 6, 11, and 12 demonstrated a significant inhibition of RPMI-7951 and T-47D cell colony formation in a soft agar clonogenic assay.