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In vitro pharmacological and toxicological effects of norterpene peroxides isolated from the Red Sea sponge Diacarnus erythraeanus on normal and cancer cells.

Eight cyclic peroxide norterpenoids, compounds 1-8, have been isolated and characterized from the Red Sea sponge Diacarnus erythraeanus, including two new norsesterterpene derivatives (3, 4). Among these metabolites, (-)-muqubilin A (5) (nine cell lines analyzed) and the new compounds 3 and 4 (seven cell lines analyzed) displayed mean IC₅₀ growth inhibitory concentrations in vitro of <10 μM, while the remaining compounds (1, 6-8) were inactive in these cancer cell lines. Compound 5 displayed no selectivity between normal and cancer cells in terms of in vitro growth inhibition. Quantitative video microscopy analysis carried out on (-)-muqubilin A-treated cells validated the data obtained by means of the MTT colorimetric assay, while flow cytometry analysis revealed ROS production but no induction of apoptosis in cancer cells.

Composition and Quantitation of Microalgal Lipids by ERETIC 1H NMR Method

Accurate characterization of biomass constituents is a crucial aspect of research in the biotechnological application of natural products. Here we report an efficient, fast and reproducible method for the identification and quantitation of fatty acids and complex lipids (triacylglycerols, glycolipids, phospholipids) in microalgae under investigation for the development of functional health products (probiotics, food ingredients, drugs, etc.) or third generation biofuels. The procedure consists of extraction of the biological matrix by modified Folch method and direct analysis of the resulting material by proton nuclear magnetic resonance (1H NMR). The protocol uses a reference electronic signal as external standard (ERETIC method) and allows assessment of total lipid content, saturation degree and class distribution in both high throughput screening of algal collection and metabolic analysis during genetic or culturing studies. As proof of concept, the methodology was applied to the analysis of three microalgal species (Thalassiosira weissflogii, Cyclotella cryptica and Nannochloropsis salina) which drastically differ for the qualitative and quantitative composition of their fatty acid-based lipids.

Antifungal Amphidinol 18 and Its 7-Sulfate Derivative from the Marine Dinoflagellate Amphidinium carterae

Two new polyketides of the amphidinol family, amphidinol 18 (AM18, 1) and its corresponding 7-sulfate derivative (AM19, 2), have been isolated from the MeOH extract of the dinoflagellate Amphidinium carterae. Structure elucidation of the two polyoxygenated molecules has been accomplished by extensive use of spectroscopic and spectrometric techniques. AM18 exhibited antifungal activity against Candida albicans at 9 μg/mL.

Development and Application of a Novel SPE-Method for Bioassay-Guided Fractionation of Marine Extracts

The biological diversity of marine habitats is a unique source of chemical compounds with potential use as pharmaceuticals, cosmetics and dietary supplements. However, biological screening and chemical analysis of marine extracts pose specific technical constraints and require adequate sample preparation. Here we report an improved method on Solid Phase Extraction (SPE) to fractionate organic extracts containing high concentration of salt that hampers the recovery of secondary metabolites. The procedure uses a water suspension to load the extracts on a poly(styrene-divynylbenzene)-based support and a stepwise organic solvent elution to effectively desalt and fractionate the organic components. The novel protocol has been tested on MeOH-soluble material from three model organisms (Reniera sarai, Dendrilla membranosa and Amphidinium carterae) and was validated on a small panel of 47 marine samples, including sponges and protists, within discovery programs for identification of immuno-stimulatory and anti-infective natural products.

Mycalol: A Natural Lipid with Promising Cytotoxic Properties against Human Anaplastic Thyroid Carcinoma Cells

The high-mobility group A (HMGA, types 1 and 2) proteins are low-molecular-weight nuclear factors that orchestrate the assembly of nucleoprotein complexes involved in gene transcription, replication, and chromatin structure. HMGAs possess oncogenic activity 2] and proteins of type 1 (HMGA1) have been correlated to cellular invasiveness and drug-resistance in human malignancies. In particular, blockage of expression of these proteins significantly enhances the responsiveness of tumor cell lines that are otherwise resistant to cytotoxic agents. Thus, phenotypic assays based on cells with reduced levels of HMGA are a possible tool for a rational search of novel compounds against tumors whose aggressiveness and resistance reduce the success of normal screening methods. Herein, we report the elucidation of the structure of mycalol (1), a novel polyoxygenated ether lipid that showed a promising in vitro specific activity against different cell lines derived from human anaplastic thyroid carcinoma (ATC), the most aggressive human thyroid gland malignancy. Mycalol was identified by a novel screening method based on the parallel use of FRO cells, which are human ATC-derived cells with high constitutive levels of HMAG1, but not HMGA2, and FRO-asHMGA1 cells, a genetically modified population of FRO cells that stably express an anti-HMGA1 antisense construct that blocks HMGA1 synthesis. The effects of extracts and fractions were measured on the paired cell lines by MTS proliferation assay. Mycalol was isolated from a chloroform extract of the sponge Mycale (Oxymycale) acerata Kirkpatrick 1907 collected along the coasts of Terra Nova Bay (Antarctica) during the Austral summer of 2005. The sponge, frozen soon after collection, was extracted with MeOH and fractionated according to a modified Kupchan method. The chloroform extract showed no activity against FRO cells up to 50 mgmL , but gave a good response (IC50 = 7.5 mgmL ) against HMGA1-silenced FRO cells (FRO-asHMGA), thus supporting the potential of a novel screening method based on HMGA-interference. Sequential steps of silica gel radial chromatography and reverse-phase HPLC (see the Supporting information) gave alkyl glyceryl ether 1 together with a number of minor compounds that are still under study.

Chemistry of the Nudibranch Aldisa andersoni: Structure and Biological Activity of Phorbazole Metabolites

The first chemical study of the Indo-Pacific dorid nudibranch Aldisa andersoni resulted in the isolation of five chlorinated phenyl-pyrrolyloxazoles belonging to the phorbazole series. Two new molecules, 9-chloro-phorbazole D and N1-methyl-phorbazole A, co-occurring with known phorbazoles A, B and D, have been characterized. Phorbazoles were found to be present mainly in the external part of the mollusc. The structures of the new compounds were determined by interpretation of spectroscopic data, mainly NMR and mass spectrometry and by comparison with the literature data. Evaluation of feeding-deterrence activity as well as in vitro growth inhibitory properties in human cancer cells was also carried out.

The green microalga Tetraselmis suecica reduces oxidative stress and induces repairing mechanisms in human cells

Green microalgae contain many active pigments such as carotenoids having antioxidant and protective activity on human cells. Here we investigate the biological activity of an ethanol/water extract of the marine green microalga Tetraselmis suecica containing high levels of carotenoids such as the xanthophylls lutein, violaxanthin, neoxanthin, antheraxanthin and loroxanthin esters. This extract has a strong antioxidant and repairing activity in the human lung cancer cell line (A549) as shown by the increased expression of dehydrocholesterol reductase-24 (DHCR24) and prostaglandin reductase 1 (PTGR1) genes and proteins. The extract also reduces prostaglandin E2 (PGE2) levels in cells damaged by H2O2 and has tissue repairing effects on reconstructed human epidermal tissue cells (EpiDermTM) indicating a potential cosmeceutical activity of this microalgal species.

Sequestered Fulvinol-Related Polyacetylenes in Peltodoris atromaculata

The Mediterranean dorid nudibranch Peltodoris atromaculata that had been collected while feeding on Haliclona fulva was shown to sequester long-chain fulvinol-like polyacetylene metabolites (compounds 2-5) from the prey. They were isolated along with previously reported bromorenierins from the diethyl ether extracts of both the mollusk and the sponge. Their structures were elucidated by NMR spectroscopy and tandem FABMS analysis. Compound 5 exhibited in vitro growth inhibitory effects against the SKMEL-28 melanoma cell line.

Dinoflagellate-Related Amphidinolides from the Brazilian Octocoral Stragulum bicolor

Benthic cnidarians are colonial marine animals that host a rich population of associated and symbiotic microorganisms. In a recent paper we described for the first time the isolation of amphidinolide P (1) from the Brazilian octocoral Stragulum bicolor. Amphidinolides and similar compounds had been previously reported only from dinoflagellates of the genus Amphidinium; thus the presence of 1 in the invertebrate opens intriguing questions on the role and occurrence of these molecules in marine ecosystems. Here we report the identification of four further amphidinolides from the same soft coral, including the known amphidinolide T1 (2) and the new analogues here named amphidinolides C4 (3), B8 (4), and B9 (5). The chemical structures have been elucidated mainly by extensive study of spectroscopic data. Cytotoxic activities of 3 and 4 were evaluated against the colon adenocarcinoma cell line HCT-116.

A new marine-derived sulfoglycolipid triggers dendritic cell activation and immune adjuvant response

Dendritic Cells (DCs) recognize infectious non-self molecules and engage the adaptive immune system thereby initiating long lasting, antigen-specific responses. As such, the ability to activate DCs is considered a key tool to enhance the efficacy and quality of vaccination. Here we report a novel immunomodulatory sulfolipid named β-SQDG18 that prototypes a class of natural-derived glycolipids able to prime human DCs by a TLR2/TLR4-independent mechanism and trigger an efficient immune response in vivo. β-SQDG18 induces maturation of DC with the upregulation of MHC II molecules and co-stimulatory proteins (CD83, CD86), as well as pro-inflammatory cytokines (IL-12 and INF-γ). Mice immunized with OVA associated to β-SQDG18 (1:500) produced a titer of anti-OVA Ig comparable to traditional adjuvants. In an experimental model of melanoma, vaccination of C57BL/6 mice with β-SQDG18-adjuvanted hgp10 peptide elicited a protective response with a reduction in tumour growth and increase in survival.