Progress of Bromophenols in Marine Algae from 2011 to 2020: Structure, Bioactivities, and Applications.

Marine algae contain various bromophenols that have been shown to possess a variety of biological activities, including antiradical, antimicrobial, anticancer, antidiabetic, anti-inflammatory effects, and so on. Here, we briefly review the recent progress of these marine algae biomaterials and their derivatives from 2011 to 2020, with respect to structure, bioactivities, and their potential application as pharmaceuticals.

Naturally Drug-Loaded Chitin: Isolation and Applications.

Naturally occurring three-dimensional (3D) biopolymer-based matrices that can be used in different biomedical applications are sustainable alternatives to various artificial 3D materials. For this purpose, chitin-based structures from marine sponges are very promising substitutes. Marine sponges from the order Verongiida (class Demospongiae) are typical examples of demosponges with well-developed chitinous skeletons. In particular, species belonging to the family Ianthellidae possess chitinous, flat, fan-like fibrous skeletons with a unique, microporous 3D architecture that makes them particularly interesting for applications. In this work, we focus our attention on the demosponge Ianthella flabelliformis (Linnaeus, 1759) for simultaneous extraction of both naturally occurring ("ready-to-use") chitin scaffolds, and biologically active bromotyrosines which are recognized as potential antibiotic, antitumor, and marine antifouling substances. We show that selected bromotyrosines are located within pigmental cells which, however, are localized within chitinous skeletal fibers of I. flabelliformis. A two-step reaction provides two products: treatment with methanol extracts the bromotyrosine compounds bastadin 25 and araplysillin-I N20 sulfamate, and a subsequent treatment with acetic acid and sodium hydroxide exposes the 3D chitinous scaffold. This scaffold is a mesh-like structure, which retains its capillary network, and its use as a potential drug delivery biomaterial was examined for the first time. The results demonstrate that sponge-derived chitin scaffolds, impregnated with decamethoxine, effectively inhibit growth of the human pathogen Staphylococcus aureus in an agar diffusion assay.

Halogen bonding for increasing efficiency in liquid-liquid microextraction: Application to the extraction of hexabromocyclododecane enantiomers in river water.

Halogen bonding (XB) was here proposed, for the first time, as a solubilization mechanism for increasing efficiency in the liquid-liquid microextraction of halogenated compounds. The approach was illustrated by the extraction of hexabromocyclododecane (HBCD) enantiomers in natural waters with a supramolecular solvent (SUPRAS) made up of inverted hexagonal aggregates of decanoic acid. The XB and dispersion interactions offered by the SUPRAS were able to extract the six HBCD enantiomers (i.e. (+)-α-, (-)-α- (+)-ß-, (-)-ß-, (+)-γ- and (-)-γ-) quantitatively (e.g. recoveries in the range 89-106%) and reach concentration factors as high as 720 without the need for solvent evaporation. HBCD enantiomers in the SUPRAS extract were directly analysed by chiral liquid chromatography coupled to tandem mass spectrometry (LCMS/MS). Quantitation limits of the method (0.09-0.9 ng L-1) were below the quality standard stablished by the European Union for HBCDs in inland surface water samples (1.6 ng L-1), and the precision, expressed as relative standard deviation (n = 6), was below 9% for all the HBCD enantiomers at concentrations within the range 50-500 ng L-1. The method was successfully applied to the enantioselective determination of HBCDs in the dissolved and the particle-bound fractions of river waters containing different concentration of suspended particles (10-57.8 mg L-1) that were spiked at two concentration levels (10 and 100 ng L-1). The results here obtained prove that XB is a valuable mechanism for the solubilisation of halogenated compounds that can effectively increase their recovery from liquid and solid samples.

Separation of hexabromocyclododecane diastereomers: Application of C18 and phenyl-hexyl ultra-performance liquid chromatography columns.

This study was performed to establish the proper liquid chromatographic conditions for the separation of hexabromocyclododecane (HBCD) diastereomers. Column selectivity towards HBCD diastereomers was evaluated for C18 and phenyl-hexyl stationary phases. First, the baseline separation of the primary HBCDs (α-, ß-, and γ-HBCD) was obtained using the ultra-performance liquid chromatography (UPLC) column with C18 stationary phase chosen in most previous studies for HBCD analysis; however, co-elution of δ- and ε-HBCD with the primary HBCD diastereomers was observed. To prevent the interference from δ- and ε-HBCD, we adopted a phenyl-hexyl UPLC column to resolve the HBCD diastereomers. The phenyl-hexyl UPLC column showed significantly different selectivity for the HBCD diastereomers compared with the C18 column, which allowed the clear isolation of δ-HBCD and ε-HBCD from the primary HBCD diastereomers. In addition, by checking the retention times of all HBCD diastereomers using both C18 and phenyl-hexyl columns, we confirmed the presence of δ-, ε-, η-, and θ-HBCDs in two technical HBCD mixtures.

Further brominated polyacetylenes with pancreatic lipase inhibitory activity from Chinese marine sponge Xestospongia testudinaria.

A new brominated polyacetylene, xestonariene I (1), along with three known related analogues (2-4), was obtained from Chinese marine sponge Xestospongia testudinaria. Its structure was determined on the basis of detailed spectroscopic analysis and by comparison with literature data. Compound 4 exhibited significant inhibitory activity against pancreatic lipase, which plays a key role in preventing obesity, with an IC50 value of 0.61 µM, being comparable to that of the positive control orlistat (IC50 = 0.78 µM).

Enantioselective Analytical- and Preparative-Scale Separation of Hexabromocyclododecane Stereoisomers Using Packed Column Supercritical Fluid Chromatography.

Hexabromocyclododecane (HBCDD) is an additive brominated flame retardant which has been listed in Annex A of the Stockholm Convention for elimination of production and use. It has been reported to persist in the environment and has the potential for enantiomer-specific degradation, accumulation, or both, making enantioselective analyses increasingly important. The six main stereoisomers of technical HBCDD (i.e., the (+) and (-) enantiomers of α-, ß-, and γ-HBCDD) were separated and isolated for the first time using enantioselective packed column supercritical fluid chromatography (pSFC) separation methods on a preparative scale. Characterization was completed using published chiral liquid chromatography (LC) methods and elution profiles, as well as X-ray crystallography, and the isolated fractions were definitively identified. Additionally, the resolution of the enantiomers, along with two minor components of the technical product (δ- and ε-HBCDD), was investigated on an analytical scale using both LC and pSFC separation techniques, and changes in elution order were highlighted. Baseline separation of all HBCDD enantiomers was achieved by pSFC on an analytical scale using a cellulose-based column. The described method emphasizes the potential associated with pSFC as a green method of isolating and analyzing environmental contaminants of concern.

Additional Insights into the Obtusallene Family: Components of Laurencia marilzae.

The obtusallenes are a significant subset of C15-halogenated acetogenins that incorporate 12-membered cyclic ethers. We have recently reported the isolation from Laurencia marilzae of 12-epoxyobtusallene IV (1) and its related α,ß-unsaturated carboxylate ester (2), both of special biogenetic relevance. Here we describe the final step of our study, the isolation of three new analogues (3-5), among these, the first bromopropargylic derivative (3) of this class of macrocyclic C15-acetogenins. The structures were elucidated by analysis of NMR and X-ray data. 12-Epoxyobtusallene IV (1), its new isomer 4, and known obtusallene IV (6) were evaluated for their apoptosis-inducing activities in a human hepatocarcinoma cell line.

Unique Polybrominated Hydrocarbons from the Australian Endemic Red Alga Ptilonia australasica.

The red alga Ptilonia australasica is endemic to Australian temperate waters. Chemical investigation of P. australasica led to the identification of four new polybrominated compounds, ptilones A-C (1-3) and australasol A (4). Their planar structures were established by extensive NMR and MS analyses. The low H/C ratio and the presence of a large number of heteroatoms made the structure elucidation challenging. The absolute configurations of 1, 2, and 4 were determined by quantum chemical ECD calculations employing time-dependent density functional theory. Ptilones A-C (1-3) show unique 4-ethyl-5-methylenecyclopent-2-enone (1 and 2) and 2-methyl-6-vinyl-4H-pyran-4-one (3) skeletons not previously reported in algal metabolites. Ptilone A displayed the most potent cytotoxicity against the human prostate cancer PC3 cells with an IC50 value of 0.44 µM and induced the PC3 cell cycle arrest in the G0/G1 phase.

Hybrid Pyrrole-Imidazole Alkaloids from the Sponge Agelas sceptrum.

A chemical investigation of the tropical sponge Agelas sceptrum from Plana Cays (Bahamas) led to the isolation of two hybrid pyrrole-imidazole alkaloids (PIAs), 15'-oxoadenosceptrin (1) and decarboxyagelamadin C (2). Herein, we report their challenging structure elucidation established by NMR and ECD spectroscopy. 15'-Oxoadenosceptrin (1) shows sceptrin merged with an adenine moiety, not yet encountered in the PIA family, whereas decarboxyagelamadin C (2) is a close derivative of agelamadins C to E recently isolated from an Agelas sp. from Okinawa.

The Alga Ochromonas danica Produces Bromosulfolipids.

Many halogenases interchangeably incorporate chlorine and bromine into organic molecules. On the basis of an unsubstantiated report that the alga Ochromonas danica, a prodigious producer of chlorosulfolipids, was able to produce bromosulfolipids, we have investigated the promiscuity of its halogenases toward bromine incorporation. We have found that bromosulfolipids are produced with the exact positional and stereochemical selectivity as in the chlorosulfolipid danicalipin A when this alga is grown under modified conditions containing excess bromide ion.

Discovery and Synthesis of Boletopsins 13 and 14, Brominated Fungal Metabolites of Terrestrial Origin.

Here we report the discovery and synthesis of complex polybrominated p-terphenyl ethers isolated from a mushroom (Boletopsis sp.) used as a traditional medicine by the Kiovi people in the highlands of Papua New Guinea. Boletopsins 13 and 14 represent the first report of polybrominated fungal metabolites to be produced by a terrestrial fungus. The synthetic method employs 2,4,4,6-tetrabromo-2,5-cyclohexadienone to achieve selective polybromination of the extended aromatic system in a selective and sequential manner.

Brominated polyunsaturated lipids with protein tyrosine phosphatase-1B inhibitory activity from Chinese marine sponge Xestospongia testudinaria.

A new brominated polyunsaturated lipid, methyl (E,E)-14,14-dibromo-4,6,13-tetradecatrienoate (1), along with three known related analogues (2-4), were isolated from the Et2O-soluble portion of the acetone extract of Chinese marine sponge Xestospongia testudinaria treated with diazomethane. The structure of the new compound was elucidated by detailed spectroscopic analysis and by comparison with literature data. Compound 3 exhibited significant inhibitory activity against protein tyrosine phosphatase 1B (PTP1B), a key target for the treatment of type II diabetes and obesity, with an IC50 value of 5.30 ± 0.61 µM, when compared to the positive control oleanolic acid (IC50 = 2.39 ± 0.26 µM).

Bioactive secondary metabolites from the Red Sea marine Verongid sponge Suberea species.

In a continuation of our efforts to identify bioactive compounds from Red Sea Verongid sponges, the organic extract of the sponge Suberea species afforded seven compounds including two new dibrominated alkaloids, subereamollines C and D (1 and 2), together with the known compounds aerothionin (3), homoaerothionin (4), aeroplysinin-1 (5), aeroplysinin-2 (6) and a revised subereaphenol C (7) as ethyl 2-(2,4-dibromo-3,6-dihydroxyphenyl)acetate. The structures of the isolated compounds were assigned by different spectral data including optical rotations, 1D (1H and 13C) and 2D (COSY, multiplicity-edited HSQC, and HMBC) NMR and high-resolution mass spectroscopy. Aerothionin (3) and subereaphenol C (7) displayed potent cytotoxic activity against HeLa cell line with IC50 values of 29 and 13.3 µM, respectively. In addition, aeroplysinin-2 (6) showed potent antimigratory activity against the human breast cancer cell line MDA-MB-231 with IC50 of 18 µM. Subereamollines C and D are new congeners of the previously reported compounds subereamollines A and B with methyl ester functionalities on the side chain. These findings provide further insight into the biosynthetic capabilities of members of the genus Suberea and the chemical diversity as well as the biological activity of these compounds.

Dactylomelane diterpenes from the sea hare Aplysia depilans.

A chemical investigation of the organic extract of the sea hare Aplysia depilans, collected off Skyros Island, Greece, yielded eight new brominated diterpenes (1-8), featuring the rare dactylomelane skeleton, together with the previously reported luzodiol (9). The structure elucidation and the assignment of the relative configurations of the new natural products were based on extensive NMR spectroscopic and MS spectrometric analyses. Compounds 1-9 were evaluated for their cytotoxic activities against five human tumor cell lines, but were proven inactive.

Activated carbons from end-products of tree nut and tree fruit production as sorbents for removing methyl bromide in ventilation effluent following postharvest chamber fumigation.

End-products of tree nuts and tree fruits grown in California, USA were evaluated for the ability to remove methyl bromide (MB) from ventilation effluent following postharvest chamber fumigation. Activated carbon sorbents from walnut and almond shells as well as peach and prune pits were prepared using different methods of pyrolysis, activation, and quenching. Each source and preparation was evaluated for yield from starting material (%, m/m) and performance on tests where MB-containing airstreams were directed through a columnar bed of the activated carbon in an experimental apparatus, termed a parallel adsorbent column tester, which was constructed as a scaled-down model of a chamber ventilation system. We report the number of doses needed to first observe the breakthrough of MB downstream of the bed and the capacity of the activated carbon for MB (%, m/m) based on a fractional percentage of MB mass sorbed at breakthrough relative to mass of the bed prior to testing. Results were based on a novel application of solid-phase microextraction with time-weighted averaging sampling of MB concentration in airstreams, which was quantitative across the range of fumigation-relevant conditions and statistically unaffected by relative humidity. Activated carbons from prune pits, prepared either by steam activation or carbon dioxide activation coupled to water quenching, received the greatest number of doses prior to breakthrough and had the highest capacity, approximately 12-14%, outperforming a commercially marketed activated carbon derived from coconut shells. Experimental evidence is presented that links discrepancy in performance to the relative potential for activated carbons to preferentially sorb water vapor relative to MB.

Identification and structure elucidation of antimicrobial compounds from Lyngbya aestuarii and Aphanothece bullosa.

Cyanobacteria are known to produce array of compounds. In an earlier report, we reported antibacterial and antifungal activities in methanolic crude extracts of laboratory grown Lyngbya aestuarii and Aphanothece bullosa isolated from Chilka Lake and local paddy field respectively. In this report the same methanolic crude extracts were subjected to TLC purification twice by altering the solvents and UV—illuminated bands bioassayed. Such UV illuminated potent bands obtained after 2nd TLC were subjected to spectroscopic analysis (UV, IR, 1H NMR and LCMS/MS). We have screened malyngolide and dragonamide C from L. aestuarii and a diterpenoid and majusculoic acid from A. bullosa. Dragonamide C and malyngolide were found to be antifungal while majusculoic acid and a diterpenoid as antibacterial. As far as our knowledge goes, this is the first ever report where fresh water A. bullosa was found to be a source of diterpenoid and majusculoic acid. Likewise, L. aestuarii was also established as a source of malyngolide and dragonamide C. This again indicated that cyanobacteria are inherently endowed with the capacity to produce metabolites according to niche and species specific manner emphasising fresh water cyanobacterial strain are as important as marine one.

An integrated approach using UHPLC-PDA-HRMS and 2D HSQC NMR for the metabolic profiling of the red alga Laurencia: dereplication and tracing of natural products.

The global metabolic profile of Laurencia crude red algal extracts was addressed by applying high-throughput analytical techniques, namely UHPLC­PDA­HRMS and 2D HSQC NMR. An integrated platform including software tools and databases, such as Xcalibur, ToxID, ACD/Labs and MarinLit, has been developed to mine the complex analytical data towards the accelerated identification of known metabolites and the detection of new natural products at the early stages of phytochemical analysis. In parallel, a searchable 'in-house' Laurencia-focused NMR database incorporating chemical structures, NMR spectroscopic data and reported biological activities has been generated. The screening strategy has been developed as a tool to prioritize the crude extracts to be further subjected to phytochemical analysis by tracing the presence of new natural products among the pool of known compounds. The successful application of this integrated methodology in the crude extract of Laurencia chondrioides led to the rapid detection of two new C15 bromoallene acetogenins (1 and 2), which were subsequently isolated and characterized.

Agelamadins C-E, bromopyrrole alkaloids comprising oroidin and 3-hydroxykynurenine from a marine sponge Agelas sp.

Three structurally unique bromopyrrole alkaloids, agelamadins C-E (1-3), were isolated from a marine sponge Agelas sp. Agelamadin C (1) possesses a hybrid structure of oroidin and 3-hydroxykynurenine connected through a dihydro-1,4-oxazine moiety. Agelamadins D (2) and E (3) are a C-9/C-10 diastereomer and a 10-epimer of 1, respectively. The structures of 1-3 were elucidated on the basis of spectroscopic analysis as well as application of a PGME method and a TDDFT ECD calculation. Antimicrobial activity of 1-3 was evaluated.

Hexabromocyclododecanes in surface sediments from Shanghai, China: spatial distribution, seasonal variation and diastereoisomer-specific profiles.

The concentrations, spatial distribution, diastereoisomer-specific profiles of hexabromocyclododecanes (HBCDs) in surface sediments from three major rivers in Shanghai as well as the seasonal variation of HBCDs were investigated. The concentrations of total HBCD diastereoisomers ranged from 0.05 to 6.87ngg(-1)dw. Significant spatial distribution of total HBCDs concentrations were observed. The mean concentration of total HBCDs followed the order of Suzhou Creek (2.00ngg(-1))>Huangpu River (1.59ngg(-1))>Yunzao Creek (0.77ngg(-1)). The concentration was a relatively low HBCDs level compared to levels measured in domestic and other parts of the world. The proportions of α-HBCD in sediment samples were generally higher than that of commercial formulations. This might be due to thermal isomerization from γ-HBCD to α-HBCD and slower degradation rate of α-HBCD compared to γ-HBCD in anaerobic conditions. The concentrations of ΣHBCD in the summer were significantly higher than those in the winter (paired t test, p<0.01). This seasonal variation could probably be attributed to a combined effect of temperature and wet deposition. Moreover, a poor and no significant correlation between ΣHBCD levels and TOC content in sediments was observed, suggesting that the spatial distributions of HBCDs were not constrained by the TOC in sediments of Shanghai.

Agelamadins A and B, dimeric bromopyrrole alkaloids from a marine sponge Agelas sp.

Two structurally unique dimeric bromopyrrole alkaloids, agelamadins A (1) and B (2), were isolated from a marine sponge Agelas sp. Agelamadins A (1) and B (2) have a structure consisting of an agelastatin-like tetracyclic moiety and an oroidin-like linear moiety in common. The structures of 1 and 2 were elucidated on the basis of spectroscopic analysis. The antimicrobial activity and cytotoxicity of agelamadins A (1) and B (2) were evaluated.