Shallow- and Deep-Water Ophiura Species Produce a Panel of Chlorin Compounds with Potent Photodynamic Anticancer Activities.

A Pacific brittle star Ophiura sarsii has previously been shown to produce a chlorin (3S,4S)-14-Ethyl-9-(hydroxymethyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid (ETPA) (1) with potent phototoxic activities, making it applicable to photodynamic therapy. Using extensive LC-MS metabolite profiling, molecular network analysis, and targeted isolation with de novo NMR structure elucidation, we herein identify five additional chlorin compounds from O. sarsii and its deep-sea relative O. ooplax: 10S-Hydroxypheophorbide a (2), Pheophorbide a (3), Pyropheophorbide a (4), (3S,4S,21R)-14-Ethyl-9-(hydroxymethyl)-21-(methoxycarbonyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid (5), and (3S,4S,21R)-14-Ethyl-21-hydroxy-9-(hydroxymethyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid (6). Chlorins 5 and 6 have not been previously reported in natural sources. Interestingly, low amounts of chlorins 1-4 and 6 could also be identified in a distant species, the basket star Gorgonocephalus cf. eucnemis, demonstrating that chlorins are produced by a wide spectrum of marine invertebrates of the class Ophiuroidea. Following the purification of these major Ophiura chlorin metabolites, we discovered the significant singlet oxygen quantum yield upon their photoinduction and the resulting phototoxicity against triple-negative breast cancer BT-20 cells. These studies identify an arsenal of brittle star chlorins as natural photosensitizers with potential photodynamic therapy applications.

Nine New Triterpene Glycosides, Magnumosides A1-A4, B1, B2, C1, C2 and C4, from the Vietnamese Sea Cucumber Neothyonidium (=Massinium) magnum: Structures and Activities against Tumor Cells Independently and in Synergy with Radioactive Irradiation.

Nine new sulfated triterpene glycosides, magnumosides A1 (1), A2 (2), A3 (3), A4 (4), B1 (5), B2 (6), C1 (7), C2 (8) and C4 (9) as well as a known colochiroside B2 (10) have been isolated from the tropical Indo-West Pacific sea cucumber Neothynidium (=Massinium) magnum (Phyllophoridae, Dendrochirotida) collected in the Vietnamese shallow waters. The structures of new glycosides were elucidated by 2D NMR spectroscopy and mass-spectrometry. All the isolated new glycosides were characterized by the non-holostane type lanostane aglycones having 18(16)-lactone and 7(8)-double bond and differed from each other by the side chains and carbohydrate moieties structures. Magnumoside A1 (1) has unprecedented 20(24)-epoxy-group in the aglycone side chain. Magnumosides of the group A (1-4) contained disaccharide monosulfated carbohydrate moieties, of the group B (5, 6)-tetrasaccharide monosulfated carbohydrate moieties and, finally, of the group C (7-9)-tetrasaccharide disulfated carbohydrate moieties. The cytotoxic activities of the compounds 1-9 against mouse spleen lymphocytes, the ascites form of mouse Ehrlich carcinoma cells, human colorectal carcinoma DLD-1 cells as well as their hemolytic effects have been studied. Interestingly, the erythrocytes were more sensitive to the glycosides action than spleenocytes and cancer cells tested. The compounds 3 and 7 significantly inhibited the colony formation and decreased the size of colonies of DLD-1 cancer cells at non-cytotoxic concentrations. Moreover, the synergism of effects of radioactive irradiation and compounds 3 and 7-9 at subtoxic doses on proliferation of DLD-1 cells was demonstrated.

Fatty acid biomarkers of symbionts and unusual inhibition of tetracosapolyenoic acid biosynthesis in corals (octocorallia).

Seven zooxanthellae-free species of octocorals (the genera Acanthogorgia, Acabaria, Chironephthya, Echinogorgia, Menella, Ellisella, and Bebryce) and two zooxanthellate octocorals (the genera Paralemnalia and Rumphella) were examined to elucidate their fatty acid (FA) composition. Arachidonic (about 40% of the total FA) and palmitic acids were predominant in all the species studied. Seven furan FA (F-acids) (up to 9.7%) were identified in the azooxanthellate octocorals. The main F-acids were 14,17-epoxy-15-methyldocosa-14,16-dienoic and 14,17-epoxy-15,16-dimethyldocosa-14,16-dienoic acids. In all specimens of Bebryce studeri, C(25-28) demospongic FA (about 20%) were identified. These FA reflect the presence of a symbiotic sponge in B. studeri and can be used as the specific markers for other corals. A significant difference (P < 0.01) between azooxanthellate and zooxanthellate corals was found for odd-chain and methyl-branched saturated FA, 18:1n-7, and 7-Me-16:1n-10; that indicated the presence of an advanced bacterial community in azooxanthellate corals. The zooxanthellate species were distinguished by significant amounts of 18:3n-6, 18:4n-3, and 16:2n-7 acids, which are proposed as the markers of zooxanthellae in soft corals. Contrary to the normal level of 24:5n-6 (9.4%) and 22:4n-6 (0.6%), unexpected low concentrations of 24:5n-6 (0.4%) accompanied by a high content of 22:4n-6 (up to 11.9%) were detected in some specimens. The presence of an unknown factor in octocorals, specific for n-6 PUFA, which inhibited elongation of 22:4n-6 to 24:4n-6, is conjectured.

Comparison of fatty acid compositions of azooxanthellate Dendronephthya and zooxanthellate soft coral species.

Ten zooxanthellae-free Dendronephthya species , twelve zooxanthellate soft coral species of the genera Sarcophyton, Lobophytum, Cladiella, Lytophyton, Cespitularia, and Clavularia, and the hermatypic coral Caulastrea tumida were examined for the first time to elucidate the fatty acid (FA) composition of total lipids. In Dendronephthya species, the main FAs were 20:4n-6, 24:5n-6, 16:0, 18:0, 7-Me-16:1n-10, and 24:6n-3 which amounted on the average to 26.0, 12.7, 12.1, 6.0, 4.8, and 4.0% of the total FA contents, respectively. For zooxanthellate soft corals, the main FAs were 16:0 (25.7%), 20:4n-6 (18.2%), 24:5n-6 (6.2%), and 18:4n-3 (5.6%), as well as 16:2n-7, which amounted up to 11.8% in Sarcophyton aff. crassum. Corals with zooxanthellae had low contents of 24:6n-3. The significant difference (p<0.01) between azooxanthellate and zooxanthellate soft corals was indicated only for 12 of 46 FAs determined. The principal components analysis confirmed that 7-Me-16:1n-10, 17:0, 18:4n-3, 18:1n-7, 20:4n-6, 22:5n-6, 24:5n-6, and 24:6n-3 are useful for chemotaxonomy of Dendronephthya. The azooxanthellate soft corals studied were distinguished by the absence of significant depth-dependent and species-specific variations of FA composition, low content of 16:2n-7, an increased proportion of bacterial FAs, predominance of n-6 FAs connected with active preying, and a high ability for biosynthesis of tetracosapolyenoic FAs.