Marine Natural Products

This review covers the literature published in 2013 for marine natural products (MNPs), with 982 citations (644 for the period January to December 2013) referring to compounds isolated from marine microorganisms and phytoplankton, green, brown and red algae, sponges, cnidarians, bryozoans, molluscs, tunicates, echinoderms, mangroves and other intertidal plants and microorganisms. The emphasis is on new compounds (1163 for 2013), together with the relevant biological activities, source organisms and country of origin. Reviews, biosynthetic studies, first syntheses, and syntheses that lead to the revision of structures or stereochemistries, have been included.


Introduction
These annual reviews of marine natural products were initiated by the late Professor D. John Faulkner in 1984 1,2 and continued by the New Zealand group since 2003. A feature of the reviews has been the inclusion of the structures for all new MNPs, and any subsequently revised structures. The number of new MNPs reported each year has steadily grown from 332 in 1984 to 1378 in this present review of the 2014 literature. This has inevitably resulted in an increased size for each review. With the everincreasing size creating difficulties for preparation of the annual review, the NPR Editorial Board suggested changing the format to focus on a selection of highlighted structures. To maintain the usual comprehensive coverage of all new and revised MNPs, we have prepared a ESI ‡ document with links associated with this review, showing all structures, along with their names, taxonomic origins, locations for collections, and biological activities. The numbers for all highlighted structures in this review (169) are shown in non-italicised bold font, while italicised numbers refer to the remaining structures in the ESI document. ‡ For structures that have their absolute congurations fully described, the compound number in the diagrams is preceded with †. In addition to the highlighted compounds in this review, we have retained the inclusion of reference to rst syntheses of MNPs, and comments on new information on ecological aspects, bioactivities or other relevant data for previously reported MNPs, all as non-highlighted material. The Reviews section (Section 2) has also been reformatted to show selected highlights, with all other reviews referenced in a section of the ESI document. ‡

Reviews
There continues to be a steady increase in the number of reviews of various aspects of MNP studies. Some of the more signicant reviews (16) are given here while a listing of the remainder (71) is given in the ESI section. ‡ A comprehensive review of MNPs reported in 2012 has appeared. 3 'Marine-sourced anticancer and cancer pain control agents in clinical and late preclinical development' have been reviewed, 4 and 'New horizons for old drugs and drug leads' were described. 5 The implications of the Convention on Biological Diversity (1999) and its Nagoya Protocol (2010) on the collection of marine genetic resources has been discussed and should be noted by all who collect marine organisms for MNP studies. 6 The putative microbial origin of sponge metabolites has been the subject of several reviews and articles. [7][8][9][10][11] Developments in chemical ecology for sh and benthic algae and invertebrates for 2010-2012 have been reviewed, with comment on the biosynthesis of bioactive MNPs by symbiotic microorganisms. 12 Polyketide biosynthesis in dinoagellates has been reviewed. 13 There have been comprehensive reviews for marine nucleosides, 14 saxitoxin, 15 and tetrodotoxin. 16 A review of 'Emerging strategies and integrated systems microbiology technologies for biodiscovery of marine bioactive compounds' provides a very useful oversight of a number of developing techniques. 17 'AlgaeBase: an on-line John Blunt obtained his BSc (Hons) and PhD degrees from the University of Canterbury, followed by postdoctoral appointments in Biochemistry at the University of Wisconsin-Madison, and with Sir Ewart Jones at Oxford University. He took up a lectureship at the University of Canterbury in 1970, from where he retired as an Emeritus Professor in 2008. His research interests are with natural products, the application of NMR techniques to structural problems, and the construction of databases to facilitate natural product investigations.
Brent Copp received his BSc (Hons) and PhD degrees from the University of Canterbury, where he studied the isolation, structure elucidation and structureactivity relationships of biologically active marine natural products under the guidance of Professors Blunt and Munro. He undertook postdoctoral research with Jon Clardy at Cornell and Chris Ireland at the University of Utah. 1992-93 was spent working in industry as an isolation chemist with Xenova Plc, before returning to New Zealand to take a lectureship at the University of Auckland, where he is currently an Associate Professor. (Hons) and PhD studies at Victoria University of Wellington. His thesis research, carried out under the guidance of Assoc. Prof. Peter Northcote, a former contributor to this review, focused on spectroscopy-guided isolation of sponge metabolites. He then carried out post-doctoral research with Mike Davies-Coleman (Rhodes University, South Africa) and Raymond Andersen (University of British Columbia, Canada) before a short role as a avour and aroma chemist at CSIRO in Adelaide, Australia. He was appointed to the faculty at his alma mater in 2009 where he is currently a Senior Lecturer.

Murray
Munro, Emeritus Professor in Chemistry at the University of Canterbury, has worked on natural products right through his career. This started with diterpenoids (PhD; Peter Grant, University of Otago), followed by alkaloids during a postdoctoral spell with Alan Battersby at Liverpool. A sabbatical with Ken Rinehart at the University of Illinois in 1973 led to an interest in marine natural products with a particular focus on bioactive compounds which has continued to this day. In recent years his research interests have widened to include terrestrial/marine fungi and actinomycetes.
Michèle Prinsep received her BSc (Hons) and PhD degrees from the University of Canterbury, where she studied the isolation and structural elucidation of biologically active secondary metabolites from sponges and bryozoans under the supervision of Professors Blunt and Munro. She undertook postdoctoral research on cyanobacteria with Richard Moore at the University of Hawaii before returning to New Zealand to take up a lectureship at the University of Waikato, where she is currently an Associate Professor. resource for algae' is an article describing a very comprehensive algal database. 18 As in previous years, the MarinLit database 19 has been updated and used as the literature source for the preparation of this present review.

Marine microorganisms and phytoplankton
Even considering the trend of recent years that many marine natural products research efforts are directed towards microorganisms, there has been a sharp upward swing in the number of new metabolites reported from marine microorganisms (677 vs. 493 in 2013). Unless otherwise stated, compounds described in this section were obtained from cultures of the named microorganisms.

Marine-sourced bacteria (excluding from mangroves)
The number of new compounds reported from marine bacteria (164) is similar to the 158 reported in 2013. A metagenomic approach has identied the gene cassette responsible for the biosynthesis of calyculin A, originally isolated from the sponge Discodermia calyx, 20 as microbial in origin, arising from Candidatus Entotheonella sp. Functional analysis of the biosynthetic pathway has shown that the end product is actually a diphosphate protoxin, phosphocalyculin A 1 rather than calyculin A, suggesting a phosphorylation/dephosphorylation mechanism for the active chemical defence of the host sponge. 21 A further diphosphate, protoxin phosphocalyculin C 2, has been isolated from D. calyx but can be assumed to have arisen from the same microbial source. Phosphocalyculin C, although potent (IC 50 ¼ 36 nM vs. P388), is 5000 times less toxic than calyculin C itself. 22 A Chinese sediment-derived Actinoalloteichus cyanogriseus 23 was the source of cyanogramide 3, an unprecedented spirocyclic alkaloid with multidrug-resistance (MDR) reversing activity. 24 An interesting dereplication strategy was utilised in the study of sponge-associated Actinokineospora sp. Principal Component Analysis (PCA), hierarchical clustering (HCA) and orthogonal partial least square-discriminant analysis (OPLS-DA) were employed to evaluate HRFTMS and NMR data of crude extracts arising from four different fermentation regimes. Statistical analysis enabled identication of the most suitable one-strainmany-compounds (OSMAC) culture conditions and extraction method resulting in isolation of the O-glycosylated angucyclines actinosporin A 4 and B 5 of which actinosporin A 4 possessed moderate activity against the causative agent of sleeping sickness, Trypanosoma brucei (T. brucei). 25 A Korean sediment-derived strain of Bacillus subtilis (B. subtilis) has yielded a variety of linear lipopeptides with differing biological properties, the gageopeptides A-D 6-9, 26 gageostatins A-C 10-12 27 and gageotetrins A-C 13-15. 28 Most are non-cytotoxic with good antibacterial activity, but better antifungal activity especially against the late blight pathogen Phytophthora capsici in the case of gageotetrins A-C. A transformation-associated recombination (TAR) cloning approach was used to capture, activate and express a 67-kb non-ribosomal peptide synthetase (NRPS) biosynthetic gene cluster from Saccharomonospora sp., resulting in isolation of the dichlorinated lipopeptide antibiotic taromycin A 16. 29 Solwaraspora sp. (ascidian, Trididemnum orbiculatum, Florida Keys, U.S.A.) produced the trialkyl-substituted aromatic acids, solwaric acids A 17 and B 18. Enrichment with 13 Clabelled glucose followed by acquisition of a 13 C-13 C COSY enabled unambiguous determination of the position of the methyl group on the phenyl ring, an approach which could be very useful for structural determination of molecules with multiple quaternary carbons. 30 Heronapyrroles A-C are nitropyrrole metabolites with a partially oxidised farnesyl chain appended that were obtained from a Streptomyces sp. 31 Biosynthetic considerations prompted the hypothesis that a mono-tetrahydrofuran-diol, heronapyrrole D might be an as yet unidentied metabolite of the bacterium. Following a putative biomimetic synthesis of heronapyrrole D 19 the metabolite was then detected in cultures of the bacterium, thus validating this approach. 32 Total synthesis of heronapyrrole C 31 has also been reported. 33 Mollemycin A 20 is a rst-in-class glycol-hexadepsipeptide-polyketide from a Streptomyces sp. (sediment, South Molle Is., Queensland, Australia) and active against certain Gram-positive and Gram negative bacteria, in addition to extremely potent antimalarial activity against drug sensitive and MDR Plasmodium falciparum (P. falciparum) clones. 34 A deep-sea strain of Streptomyces, previously a source of spiroindimicins A-D, 35 [173][174][175][176] and a bio-inspired total synthesis of the indole sesquiterpenoid sespenine 177 completed. 178

Marine-sourced fungi (excluding from mangroves)
Studies of fungi continue to rise with 318 new compounds reported in 2014 compared to 223 in 2013. Sponge-associated Aspergillus similanensis yielded two isocoumarin derivatives 204 and 205, and a deacetyl analogue of chevalone C, 179 chevalone E 206, in addition to pyripyropene S 207. 180 While chevalone E 206 itself did not display signicant antibacterial activity, it did exhibit synergy with the antibiotics oxacillin and ampicillin against MRSA. 181 Ultrasonication of Aspergillus versicolor spores led to the isolation of a mutant with neomycin resistance from which six metabolites not present in the parent strain, including 208 were obtained. Although several patents exist for synthesis of the planar structure of this compound, [182][183][184] this is the rst natural product (NP) isolation and establishment of stereochemistry.
Cyclo(D-Tyr-D-Pro) was also claimed as a rst NP isolation but this has previously been obtained from both terrestrial 185 and marine 186 sources. 187 Lumazine peptides, penilumamide B-D 209-211 and a cyclic pentapeptide, asperpeptide A 212 were obtained from a gorgonian-derived Aspergillus sp. The presence of the sulfone penilumamide 188 and the derived sulfoxide penilumamide C 210 led to speculation that these compounds were derived from oxidation of a putative metabolite containing a methionine residue. This led to a feeding experiment with Lmethionine resulting in isolation of the sulde, penilumamide B 209. Yields of penilumamide B 209 and penilumamide increased with increasing concentration of L-methionine and when penilumamide B 209 was exposed to air, penilumamide was detected aer a few days whilst penilumamide C 210 was formed several days later. 189 Of the two prenylated hydroquinone derivatives 213 and 214 obtained from a gorgonian-derived Aspergillus sp., 214 exhibited very potent activity against respiratory syncytial virus (RSV). 190 These two metabolites differ only in the conguration of a methyl group on a cyclohexane ring yet given that 214 is an extremely potent anti-RSV agent whilst its epimer 213 is completely inactive, indicating the importance of the conguration of this ring for anti RSV-activity.
So-coral associated Chondrostereum sp. has previously been reported to produce hirsutane-framed sesquiterpenes. [191][192][193] Cultivation of the fungus in a medium with glycerol as the carbon source led to the isolation of the sesquiterpenes chondrosterin I 215 and J 216 of which chondrosterin J 216 displayed potent activity against HTCLs. 194 Chemical epigenetic modication of Cochliobolus lunatus (sea anemone Palythoa haddoni) with inhibitors of histone deacetylase (HDAC) resulted in isolation of two brominated 14-membered resorcylic acid lactones 217 and 218, but only in the presence of an HDAC inhibitor. 195 The chromones engyodontiumone A-H 219-226 and the phenol derivatives 227-229 were derived from deep-sea derived Engyodontium album. Of these, engyodontiumones E-G 223-225 were obtained as racemates. 196 The known polyketide aspergillus one B 197 was also isolated and was a potent inhibitor of settlement of Balanus amphitrite (B. amphitrite) larvae. 196 Fermentation of a lamentous fungus of the Eurotiomycetes class (ascidian, Lissoclinum patella, Papua New Guinea) produced the pentacyclic oxazinin A 230, derived from a combination of benzoxazine, isoquinoline and pyran rings. Oxazinin A 230 occurred as a racemate and was antimycobacterial. 198 Neosartorya pseudoscheri (starsh Acanthaster planci, Hainan, China) produced different suites of metabolites when cultured in different media. When cultured in glycerolpeptone-yeast extract (GlyPy), the diketopiperazines neosartin A 231 and B 232 were produced along with six known diketopiperazines and a precursor alkaloid but when fermented in glucose-peptone-yeast extract (GluPy), a tetracyclic-fused alkaloid neosartin C 233 was produced along with a known meroterpenoid and ve known gliotoxin analogues 234-238, obtained here for the rst time as NPs. 199   Metabolites 245-254 were also obtained from the genera Acremonium, Arthrinium, Ascotricha and Astrocystis. [204][205][206][207] As usual, a large number of metabolites were obtained from species of the Aspergillus genus. Two indole diterpenoids 255 and 256, and an isocoumarin 257 were obtained from A. avus. The known compounds b-aatrem, 208 paspalinine 209,210 and leporine B 211 were isolated and leporine A 212 was prepared from the last of these. Congurations were established for each as 258-261 respectively and b-aatrem and leporine B were isolated for the rst time as MNPs. 213 Other metabolites isolated from the genus Aspergillus include 262-327 (the last obtained from co-culture with an unidentied bacterium).  New metabolites 328-371 were also obtained from the genera Beauveria, Cladosporium, Cochliobolus, Curvularia, Dendrodochium, Diaporthaceae, Dichotomomyces, Emericella and Eurotium. [238][239][240][241][242][243][244][245][246][247][248][249][250] A pyrrolidinoindoline diketopiperazine dimer, cristatumin E, isolated from Eurotium herbariorum, 251 appears to be identical to the previously reported eurocristatine. 252 The genera Hansfordia, Humicola, Isaria, Neosartorya, Nigrospora, Paecilomyces and Paraconiothyrium also yielded the new metabolites 372-399. 179,253-258 The genus Penicillium was the source of many other metabolites 400-461. [259][260][261][262][263][264][265][266][267][268][269][270][271][272][273][274][275][276][277][278] Other genera to yield new the metabolites 462-520 were Pseudallescheria, Spicaria, Spiromastix, Stachybotrys, Talaromyces, Trichoderma, Xylaria and Xylariaceae, 279-291 while 521 was obtained from a strain of the order Xylariales and also synthesised. 292 A number of syntheses of fungal metabolites have resulted in structural revisions of the natural products, including syntheses of (À)-protubonine A 522, (À)-protubonine B 523 (Aspergillus sp.) 293,294 and (+)-cristatumin C 524 (Eurotium cristatum). 295,296 Synthesis of the racemate of oxalicumone C (Penicillium oxalicum) 297 followed by resolution by chiral HPLC and examination of experimental and calculated ECD data, resulted in the conguration of the natural product being assigned as (S)-525. 298 Culture of a strain of Aspergillus clavatus isolated from the hydrothermal vent crab Xenograpsus testudinatus in the presence of the abiotic stress agent and hydrothermal vent uid component zinc (as zinc sulfate), elicited production of a known synthetic cyclic peptide 299 that was isolated for the rst time from a natural source and named as clavatustide C 526. The fungus did not produce clavatustide C 526 when cultured in the absence of zinc. 300 Pericosine E 301 (Periconia byssoides) occurs as an enantiomeric mixture in nature and synthesis of the preferred enantiomer (À)-pericosine E 527 has been reported. 302 First syntheses of a number of fungal metabolites achieved include those of secalonic acids A 303 (Paecilomyces sp.) 304 and D 305 (Gliocladium sp.) 306,307 (AE)-sorbiterrin A 308 (Penicillium terrestre), 309 (À)-auronamide C 310 (Penicillium aurantiogriseum), 311 calcaripeptides A-C 312 (Calcarisporium sp.), 313 (À)-aspergilazine A 314 (Aspergillus taichungensis), 315 328 The benzaldehyde derivative isotetrahydro-auroglaucin 329,330 has been shown to exhibit anti-inammatory activity, inhibiting the NF-kB pathway through suppressing production of both pro-inammatory mediators and cytokines. 331 Oxirapentyns A 332,333 and B 333 exhibited growth stimulatory effects on seedling roots of barley and wheat 334 while isoechinulin A 335,336 was a strong inhibitor of settlement of larvae of the barnacle B. amphitrite. 337 Methylpenicinoline 338 also displayed anti-inammatory activity, suppressing expression of pro-inammatory mediators through the NF-kB and MAPK pathways. 339 Heavy metal stress of two strains of hydrothermal vent fungi (Aspergillus sclerotiorum and A. clavatus) induced biosynthesis of metabolites that were not produced under normal culture conditions: A. sclerotiorum produced aspochracin 340,341 when stressed with copper and produced stephacidin A 342 and notoamides B 342 and F 343 under normal culture conditions whilst A. clavatus produced the acetophenone derivative clavatol 344,345 under stress conditions and deoxytryptoquivaline 346 and tryptoquivaline A 346 in metal-free culture. 347

Fungi from mangroves
There has been a considerable increase in the number of new metabolites reported from mangrove-associated fungi (108 in 2014 vs. 75 in 2013), with the majority coming from endophytic species. Co-culture of Alternaria and Phomopsis species led to isolation of three cyclic peptides 528-530, all of which exhibited signicant activity against a range of plant pathogenic fungi, 348,349 whilst co-culture of two brown alga (Sargassum)derived Aspergillus species also produced a cyclic peptide, psychrophilin E 531. 350 Aspergillus avipes was the source of the aromatic butyrolactones, avipesin A 532 and B 533 and the previously synthesised 534 351 and 535, 352 of which avipesin A 532 exhibited moderate to good antibacterial activity. Unlike penicillin, it was able to penetrate the biolm matrix to kill live bacteria inside mature Staphylococcus aureus biolm. 353 An endophytic Diaporthe sp. was the source of diaporine 536, an unprecedented symmetric polyketide which induces conversion of tumour associated macrophages from the M2 to the M1 phenotype in both cellular and animal models. 354 The polyketides dothiorelone F 537 and I 538 were obtained from endophytic Dothiorella sp. along with three known analogues. 355 Of these analogues, dothiorelone G 356 is actually the same as the previously reported cytosporone R, 357 also obtained from a mangrove associated species (Leucostoma persoonii). Peniphenones A-D 539-543 were obtained from Penicillium dipodomyicola. Of these, peniphenone A 539, 540 occurs as a racemate and was separated into its enantiomers by chiral HPLC while peniphenones B 541 and C 542 strongly inhibited Mycobacterium tuberculosis protein tyrosine phosphatase B (MptpB). 358 The polyketides 544 and 545 were obtained from co-culture of mangrove soil derived Penicillium sp. with the sediment derived bacterium Streptomyces fradiae and were not produced in discrete bacterial and fungal control cultures, suggesting the activation of silent gene clusters by co-cultivation. These also occurred as a racemate and were separated by chiral chromatography into 544 359 a known terrestrial fungal metabolite (9R,14S)-epoxy-11-deoxyfunicone, but now isolated as a rsttime MNP, and the enantiomer (9S,14R)-epoxy-11-deoxyfunicone 545. 360 An endophytic Penicillium sp. was the source of a phenyl ether derivative 546 and a spiroax-4-ene-12-one derivative 547 with the spiroax-4-ene-12-one derivative 547 more potent to the MG-63 cell line with in vivo activity and signicant inhibition of human osteosarcoma in nude mice upon oral administration. 361 The prenylated phenols vaccinol A-G, 548-554 and the naphthalene derivative vaccinal A 555 were isolated from Pestalotiopsis vaccinii. Vaccinal A 555 exhibited potent COX-2 inhibition. 362 Other genera or families of fungi associated with mangroves to yield the new metabolites 556-635 were Acremonium, Alternaria, Daldinia, Guignardia, Penicillium, Pestalotiopsis, Phoma, Phomopsis, Pseudolagarobasidium, Rhytidhysteron, Stemphylium and Xylariaceae. 363-388

Cyanobacteria
There has been an increase in the number of new metabolites reported from cyanobacteria since 2013, but the total numbers are still low overall (20 in 2014 vs. 9 in 2013), seemingly continuing an overall downward trend. Typical of the phylum, the vast majority of metabolites reported were peptidic in nature. Although not peptidic, yoshinone A 636 and the diastereoisomers yoshinone B1 and B2 637 were isolated from Leptolyngbya sp. Yoshinone A 636 inhibited differentiation of 3T3-L1 cells into adipocytes without accompanying cytotoxicity suggesting potential as an anti-obesity lead. 389 A cyanobacterial assemblage, consisting mostly of Lyngbya sp. (now renamed as Moorea sp.) yielded the dolastatin 13 analogue, kurahamide 638, a strong inhibitor of the proteases elastase and chymotrypsin 390 and the acetylenic lipopeptide kurahyne 639 an inhibitor of the HeLa cell line and inducer of apoptosis. 391 Mooreamide A 640 is a cannabinomimetic lipid obtained from Moorea bouillonii and is the most potent marine-derived inhibitor of the neuroreceptor CB 1 reported to date. 392 Two new aplysiatoxin analogues, 3-methoxyaplysiatoxin 641 and 3methoxydebromoaplysiatoxin 642 were isolated from Trichodesmium erythraeum and 642, along with the co-isolated known debromo analogues debromoaplysiatoxin 393 and anhydrodebromoaplysiatoxin, 394 were inhibitors of the Chikungunya virus. 395 Other new metabolites 643-655 were obtained from the genera Anabaena, Lyngbya, Moorea, Oscillatoria and Symploca. [396][397][398][399][400][401][402] The absolute congurations of coibacins A and B 403 have been determined as 656 and 657 respectively by total synthesis. 404 414 have also been achieved. Total syntheses of the proposed structures for itralamide B 415 and coibamide A 416 have indicated that the structures of these natural products require revision. 417,418 Grassypeptolides A-C 419 were shown to selectively inhibit the dipeptidyl peptidase (DPP8) protease and molecular docking studies indicated that grassypeptolide A binds to the enzyme at two different sites. 420 Gallinamide A 421 was shown to be a potent and selective inhibitor of the human cysteine protease cathepsin L. 422 A genome mining approach was utilised to identify three proteusin rSAM epimerases, enzymes which install multiple D-amino acids in genetically encoded peptide chains, from three strains of cyanobacteria including a marinederived Pleurocapsa strain. 423 A genome mining approach was also used to show that LanA peptides, linear precursor peptides of lanthionine-containing peptides (lanthipeptides), 424 are highly diverse among different systems and that closely related lanthipeptide synthetases can be associated with quite different substrate sets. 425

Dinoagellates
The number of new metabolites reported from dinoagellates has remained about the same as for 2013 with 19 compounds reported in 2014. The linear polyketide, amphirionin-4 658, from an Amphidinium species displayed very potent and selective growth promotion activity on murine bone marrow stromal ST-2 cells. Feeding experiments with 13 C single and double labelled acetate indicated that polyketide 658 comprises a linear C22 chain with two irregular C1 sites (in the terahydrofuran moiety) and four C1 branches. 426 Dinophysis acuminata was the source of the macrolide acuminolide A 659 which was non-toxic to human tumour cell lines (HTCLs) but a potent stimulator of actomyosin ATPase activity. 427 Belizentrin 660, a 25-membered macrolactam obtained from Prorocentrum belizeanum is the rst member of its class of polyunsaturated and polyhydroxylated macrocycles and displayed potent effects on neuronal network integrity in cerebellar cells, ultimately resulting in cell death. 428 Other genera of dinoagellates from which the new metabolites 661-675 were isolated include Amphidinium, Azadinium, Karenia and Vulcanodinium. [429][430][431][432][433][434][435][436] Tentative structures were assigned to ovatoxin-g 676 and an isomer of palytoxin, the so-called isobaric palytoxin. 437 The absolute conguration of amphidinin A 438 was determined as 677. 439 Synthesis of genetically predicted saxitoxin intermediates with identication and quantication in the dinoagellate Alexandrium tamarense and the cyanobacterium Anabaena cicinalis supports the genetically proposed biosynthetic route 440 to saxitoxin. 441 Metabolomic and proteomic analyses indicated that Karenia brevis exhibited allelopathy against two competitor diatoms Asterionellopsis glacialis and Thalassiosira pseudonana. The former, which co-occurs with K. brevis, exhibited somewhat more robust metabolism while in the latter, energy metabolism was disrupted and cellular protection mechanisms were impeded. 442 Other studies examined the biosynthesis of brevetoxin, 443,444 brevisamide 445,446 and the genetics of toxin production in Alexandrium catenella. 447

Green algae
Research into green algal chemistry continues at low ebb with just 13 relevant articles published and just three new compounds published for 2014. The new compounds were the racemosines A-C 678-680, 448,449 bisindole alkaloids isolated from Caulerpa racemosa which are biosynthetically related to the well-established green algal metabolite caulerpin, also from a Caulerpa sp. 450 Caulerpin featured in studies of antinociception mechanisms 451 and the antituberculosis activities of caulerpin and synthetic analogues. 452 Included in the green algal literature for 2014 was a study on the effect of natural lycopene (E/Z mixture) on a human prostate cancer cell line, 453 while a thought-provoking paper tackled a problem all marine natural product chemists should be alert tothe misuse of taxonomic descriptors and the implications that this has. Misuse of such descriptors appears to be a particular problem with marine algae. 454

Brown algae
The output of new compounds (17) from brown algae in 2014 was again relatively low and although dominated by terpenoid chemistry saw the emergence of a new class of brown algal metabolite. This was the characterisation of a 1-deoxysphingoid, 3-epi-xestoaminol C 681, from a New Zealand collection of Xiphophora chondrophylla following a M. tuberculosis-guided fractionation. A genome-wide screening against a library of non-essential gene deletion mutants of Saccharomyces cerevisiae established the cellular processes that 681 disrupted. 455 Some 27 diterpenoids ranging across six classes were isolated from a Chinese collection of Dictyota plectens. These included seven new diterpenes, belonging to the dolabellanes 682-684, prenylated guaianes 685-687, and a xenicane 688, 19 known analogues and an ethoxylated artifact as well. 456 Three new diterpenoids, a dolabellane 689 a xenicane 690, and a prenylated guaiane 691 with ve previously characterised compounds were characterised from Mediterranean collections of several Dictyota spp. 457 Three new, 692-694, and three known dolabellane diterpenoids, 458,459 were isolated from a Brazilian D. pfaffi, and a single crystal X-ray analysis established the absolute conguration of the known 10,18-diacetoxy-8-hydroxy-2,6-dolabelladiene. 458,460 The meroterpenoids sargachromanol Q 695 and R 696 along with the known sargachromanol J 461 resulted from re-examination of the original extract from Sargassum siliquastrum, 462 while thunberol, 697, is the only new sterol of ve isolated from the Chinese Sargassum thunbergii. 463 Typical brown algal phlorotannins such as eckol and dieckol have stimulated research into a wide range of topicsheme oxygenase-1 expression, 464 oxidative stress, 465 anti-adipogenic activity, 466 anti-HIV-1 467 and antibacterial activity. 468 Among the studies on the anti-inammatory properties of phlorotannins 469,470 was the synthesis of a rhodamine-labelled dieckol. Confocal laser microscopy determined that the labeled dieckol was mainly located in the endoplasmic reticulum and studies showed that the anti-inammatory activity of the conjugate was considerably greater than that of dieckol itself. 471 The potential therapeutic properties of fucoxanthin and derivatives have been studied, 472,473 as have those of polyphenols such as octaphlorethol, [474][475][476] and mono-and diacylglycerols. 477

Red algae
There continues to be a marked variation in the number of new compounds obtained from red algae and reported each year, with 42 for 2014 compared to 9 for 2013. There was the usual range of structural types with glycolipids 698 478  A synthesis 491 of the brominated sesquiterpene aldingenin C 492 showed that the original structure was incorrect, and it was suggested that aldingenin C was probably the known compound caespitol. 493 A synthesis 494 of the proposed structure of prevezol B 495 has shown that the original structure was incorrect. Of the $70 polyhalogenated acyclic monoterpenes isolated from Plocamium spp., surprisingly none had been synthesised until the use of a broadly applicable approach generated four of the naturally occurring compounds and a number of analogues. 496 An asymmetric total synthesis of (+)-bermudenynol 497 has been accomplished, 498 while a total synthesis of the snyderane (+)-luzofuran 499 has also been made. 500 The anti-inammatory potential of 5b-hydroxypalisadin B 501 from Laurencia snackeyi has been demonstrated in LPS-stimulated RAW 264.7 macrophages 502 and LPS-induced zebrash embryo. 503

Sponges
With 283 new structures reported from the phylum Porifera in 2014, sponges have returned again to be a dominant source of new bioactive metabolites, although the growing realisation that microbial symbionts are the real producers of "sponge" specialised metabolites highlights the need for more detailed metagenomic and biosynthetic analyses of sponge matrices.
Sphingoids 740 504 and 741, 505 taurinated fatty acids 742-745 506,507 and a large number of polyacetylenes 746-765 [508][509][510][511][512][513] have been reported from the genera Callyspongia, Coscinoderma, Echinoclathria, Petrosia, Placospongia, Siphonochalina and Xestospongia. The synthesis of a series of miyakosyne A (Petrosia sp.) 514 diastereomers followed by RP-HPLC separation at À56 C has revealed that the natural product 766 is actually a mixture of two stereoisomers in $96 : 4 (14R) : (14S) ratio. 515 A comprehensive blend of synthesis with NMR, IR and vibrational circular dichroism (VCD) spectroscopy has allowed for the relative congurational assignment of the C-36-C-42 segment of hemicalide 767 (Hemimycale sp.), as well as securing the absolute conguration of C-42. This potent (sub-nM) inhibitor of mitosis was sourced from a deep water sponge collected at the Torres Islands, Vanuatu, but to date has only been reported in a patent. 516,517 A series of peroxides 768-773, 518 halogenated alkenes 774 and 775, 519 and an amide 776 520 have been reported from Plakortis simplex, Dysidea sp. and Anoxycalyx (Scolymastra) joubini, respectively. Sponges continue to be a prolic source of peptide natural products. Reisolation of cyclolithistide A from a deep sea Discodermia japonica (À200 m, Sagami Bay, Japan), originally reported in 1998, 521 allowed in-depth LC-MS/MS and Marfey's analyses that necessitated a revision of the amino acid sequence and congurations as shown here 777. 522 The simple substitution of a side-chain Asp for hydroxy-Asp in pipecolidepsins A 778 and B 779 (Homophymia lamellosa, Saint Marie Is., Madagascar) resulted in a greater than ten-fold increase in activity against three HTCLs. 523 The synthesis of 778 had already been reported in 2013. 524 The stellatolides A-G 780-786 are a family of cyclodepsipeptides from a Madagascan Ecionemia acervus. The full stereochemical analyses of several variants were established by Marfey's analysis, and the solid-phase peptide total synthesis of 780 was also achieved. The unexplained racemisation of L-Thr during the Marfey's analysis is a salient warning to all experimentalists using this technique. Several of the stellatolides were cytotoxic in the nM range against three HTCLs. 525 The genera Asteropus, Discodermia, Pipestela, Reniochalina, Stylissa, Suberites and Theonella have also yielded a large number of peptides 787-806. 526-532 Callyspongiolide 807 is an unusual carbamate macrolide with an unprecedented conjugated dienyne side chain isolated from Callyspongia sp. (Ambon, Indonesia). Evaluation of 807 against three HTCLs indicated potent cytotoxicity (IC 50 ¼ 60-320 nM). Notably, the viability of cell lines treated with callyspongiolide was not affected by QVD-OPh, a known caspase-inhibitor, suggesting the test compound induces cellular toxicity in a caspase-independent manner. 533 The total synthesis of halichondrin A has been accomplished. Halichondrin A is a "phantom" natural product, viz. it is the only member of the norhalichondrin/halichondrin/homohalichondrin family not to have been detected from a natural source, typically Halichondria okadai, 534,535 even though concerted efforts have been made to try and detect this compound for more than 20 years. Use of the synthetic material to guide studies searching for the presence of halichondrin A in legacy extracts of H. okadai were unsuccessful. 536 A diketopiperazine 808, 537 a 3-alkylpiperidine 809, 538 seven manzamine-type alkaloids 810-816 539,540 and seven 3-alkylpyridines 817-823 541 have been isolated from the sponge genera Callyspongia, Acanthostrongylophora and Topsentia. Axinyssa aculeata (Okinawa) was the source of protoaculeine B 824 that is a putative novel N-terminal residue for the peptidic toxins aculeines A-C. The structure of this polyamine compound, including identifying its attachment to the peptide toxins themselves, was determined from detailed synthetic and mass spectrometric studies. 542 Other indole 825-829 543,544 and b-carboline 830-837 544-546 alkaloids were reported from Hyrtios, and Luffariella sponges. A biomimetic total synthesis of (AE)-dictazole B, isolated from the shallow water sponge Smenospongia cerebriformis, 547 has been performed using articial light to facilitate the [2 + 2] photochemical cycloaddition reaction between aplysinopsin monomers, under the presumed high local concentration of the parent reactant that would be found in the relevant sponge organelle. This biomimetic route is a viable alternative to the [4 + 2] Diels-Alder cycloaddition that has been presumed to take place in the biosynthesis of the cycloaplysinopsins. 548 Fieen new aaptamine alkaloids 838-852 [549][550][551] have been reported from Aaptos sponges while a new polycyclic aromatic alkaloid 853 came from an Ancorina sp. 552 An unbiased screen of a library of 480 sponge extracts identied the surprisingly simple compound girolline 553 from Stylissa carteri (Mont Pass, Pohnpei, Micronesia) as a potent inhibitor of signalling of MyD88-dependent and -independent Toll-Like Receptors (TLR) 2, 3, 4, 5 and 7, reducing cytokine production in peripheral blood mononuclear cells and macrophages, therefore imbuing the molecule with potent anti-inammatory activity. In-depth chemical genetics proling identied elongation factor 2 as the molecular target of girolline implying inhibition of protein synthesis as its mode of action. 554 Guanidine-derived alkaloids 854-863 were reported from Monanchora pulchra, 555 Acanthella cavernosa 556 and Biemna laboutei, 557 while several brominated hymenialdisine-type 864-867 and oroidin/oroidin dimer-type 868-877 came from Callyspongia sp. 558 and Agelas sp., 559-561 respectively. A highly scalable, multigram synthesis of axinellamines A and B (Axinella sp.) 562 and analogues has been achieved. Evaluation of the antibacterial properties of racemic synthetic products highlighted their potent inhibition of both Gram positive and negative bacteria (MIC 0.5-8 mg mL À1 vs. eight bacterial cell lines), even though the initial isolation study reported minimal activity. Further detailed investigation could not determine a specic mode of action but did suggest secondary membrane-disruption as the likely route to activity. 563 The comprehensive total synthesis of sceptrin, ageliferin and massadine congeners via single electron cycloaddition reactions has several wide-ranging consequences. Firstly, it necessitates the revision of the absolute conguration of sceptrin and its brominated analogues 878-880 [564][565][566] and ageliferin and congeners 881-883 567 as shown here, based upon X-ray crystal structures. Moreover, the massadines had been thought to be rearrangement products from the ageliferins, 568,569 however the absolute congurations of the two series of compounds are antipodal, hence indicating enantiodivergent biosynthetic pathways in vivo to these bromopyrrole alkaloids and invalidating the so-called "Scheuer-hypothesis" for their formation. The total syntheses of 932 and two analogues were achieved using the rarely applied silver-catalysed Sato photochemical coupling, leading to a short and efficient synthesis of the natural product. Comparative analysis of 932 and its analogues showed that the 3-OH functional group is highly detrimental for IDO-inhibitory activity. 585 A merotetraterpenoid 933 has been reported from Sarcotragus spinosulus. 586 Similarly, Axinyssa, Dysidea, Ircinia and Topsentia sponges were the sources of a number of sesquiterpenoids 934-964, 587-591 some of which had been cleaved, while diterpenoids were obtained from Diacarnus megaspinorhabdosa 965-970, 592 Spongia sp. 971-973 593 and Darwinella oxeata 974-982. 594  Cinanthrenol A 1027 is the rst example of a phenanthrenecontaining steroid. It was sourced from Cinachyrella sp. collected by dredging at À160 m (Oshima-Shinsone, Japan) where the unique phenanthrene-uorescence signature was used to select the sponge from amongst detritus. As well as moderate activity against various HTCLs, 1027 was a potent estrogen-receptor binder, displacing estradiol in a competitive manner with IC 50 ¼ 10 nM, as well as altering estrogenresponsive gene expression. 608 A series of triterpenoids 1028-1032 have been reported from Jaspis stellifera 609 and Siphonochalina siphonella. 610 An articial "sponge" has been developed for the non-destructive concentration of marine natural products in sensitive marine ecosystems. A pump is used to suction sponge-matrix particles that are ltered into a hollow-bre bioreactor. This is used to cultivate microbial symbionts that produce secondary metabolites which are, in turn, trapped on reversed-phase cartridges for concentration. The articial sponge system was used in three expeditions across 11 locations, two in the Pacic and one in the South China Sea, with the deployment at Pulau Lakei (Sarawak, Malaysia) yielding jasplakinolide, jasplakinolide B and C in a 18 : 1 : 1 ratio, the same proportions as found from Jaspis splendens collected near Vanuatu. 611,612 Metagenomic analyses continue to rise in their importance to sponge natural product biosyntheses. Detailed metagenomic proling of the "High Microbial Abundance" (38-57% microbial biomass) sponge Plakortis halichondrioides (Little San Salvador Is., Caribbean Sea) found characteristic gene sequences of poribacteria (a bacterial phylum found exclusively in sponges) but the known supA and swfA biosynthetic clusters observed were absent from the poribacteria genomes. Further investigation showed similarity between these clusters and protist type I polyketide synthetase (PKS) genes hence protists could be a previously overlooked reservoir of novel bioactive metabolites. 613 A comprehensive metagenomic survey of the microbial community associated with the metabolite-rich sponge Theonella swinhoei has uncovered a new bacterial genus, Entotheonella. This genus is a member of the new candidate phylum Tectomicrobia and appears to be a widespread sponge endosymbiont across many diverse sponge species. Moreover, biosynthetic gene cassettes suitable for production of many "Theonella swinhoei" NRPS and PKS compounds, including the onnamide, 614 polytheonamide, 615 keramamide 616 and cyclotheonamide 617 classes, have been identied in the microbial genome. 8,9 A metagenomic approach identied the gene cassette for calyculin A (Discodermia calyx) 20 biosynthesis as being microbial in origin, as described in 3.1 Bacteria, and resulted in the identication of two diphosphate protoxins (see 1 and 2), 21,22 again underscoring the importance of symbiotic microbial communities in the production of "sponge" metabolites. The microtubule-stabilising mode of action of laulimalide 618 and peloruside A 619 has been established through the use of highresolution crystal structures of the compounds bound to tubulin, 620 while aaptamine 621 has been shown to have protective effects against cisplatin-induced kidney damage, 622 and also prevents photoageing. 623 A polybrominated diphenylether 624 (Dysidea granulosa) has been shown to inhibit hepatitis C nonstructural protein NS3 helicase, a validated antiviral target. 625 The four bromotyrosine compounds ianthelliformisamine A-C (Suberea ianthelliformis) 626 and spermatinamine (Pseudoceratina sp.) 627 were selective and potent inhibitors of human carbonic anhydrase IX (IC 50 ¼ 0.20-0.36 mM), with implications for maintaining cellular pH homeostasis and hence application in a variety of disease states. 628 The sesterterpenoid heteronemin (Hyrtios sp.) 629 inhibited trans-activation response DNA-binding protein 43 kDa (TDP-43), a key factor of several neurodegenerative states. Heteronemin binds to TDP-43 (k i ¼ 270 nM) and altered the aggregation state and localisation of this important neurochemical target. 630 The rst total syntheses of an all-(Z) octadeca-pentene

Cnidarians
The number of new compounds reported from cnidarians in 2014 (201) is similar to the previous decadal average. The importance of chemical cues in coral reef remediation have been highlighted in a study that found that coral and sh juveniles were repelled from reefs that were overshed and seaweeddominated. 682 A method of recovery of such degraded habitats was proposed to involve reduction in shing harvest of critical species of herbivorous shes. The chemistry of both hard and so corals are dominated by terpenesalkaloids are only rarely isolated. In 2014 there were seven examples of alkaloids isolated from so corals, comprised of an unusual diketopiperazine 1040 from Menella kanisa, 683 and six tetraprenylated purines, malonganenone L-Q 1041-1046, from Echinogorgia pseudossapo. 684 In contrast, cnidarians of the order Zoantharia (zoanthids) were the sources of ve new parazoanthine congeners 1047-1051 (Parazoanthus axinellae). 685 These structures were established using a metabolomics model, with LCMS/MS fragment analysis being used to dene structures and LC-ECD employed to assign absolute conguration (to 1047, 1048 and 1050).
Other zoanthamine analogues, 1052-1058, were isolated from Zoanthus sp. 686,687 and Zoanthus kuroshio 688 as well as a new 42-hydroxypalytoxin diastereomer 1059 from Palythoa tuberculosa. 689 Previous studies of an extract of Palythoa toxica identi-ed the major toxin to be a 42-hydroxy analogue of palytoxin 1060 however the conguration at positions C-41 and C-42 remained unresolved. 690 More recent J-based analysis has dened the conguration at these stereocentres as (41S) and (42S). The same study also established the absolute conguration of a related palytoxin analogue isolated from P. tuberculosa as being (42S)hydroxy-(50S)-palytoxin 1059. The relative affinity of palytoxin, 1059 and 1060 towards a mouse anti-palytoxin monoclonal antibody identied palytoxin to have the most potent K D (nM), while 1059 and 1060 were approximately one and three orders of magnitude less potent respectively. Similar relative levels of cytotoxicity towards keratinocytes were also observed, with palytoxin being the most potent. Palytoxin is known to be capable of disrupting mechanisms of cellular ion homeostasis: NMR studies have dened the C-25-C-33 and C-47-C-53 fragments of palytoxin as being involved with Ca 2+ coordination. 691 A series of butenolide and cyclopentenones 1061-1072 were reported from Subergorgia suberosa and Sinularia sp. so corals. [692][693][694] A comprehensive investigation of a Bahamian collection of Pseudopterogorgia rigida identied a chamigrane 1073 and seven bisabolanes 1074-1079 as new sesquiterpenes. 695 The study also reported an extensive number of cometabolites 1080-1083 that had been previously reported from terrestrial plants or as semi-synthetic derivatives 1084-1090. Other sesquiterpenes 1091-1110 have been reported from Rumphella antipathies, [696][697][698] Lemnalia philippinensis, 699 Menella kanisa, 700 Nephthea erecta, 701 an unidentied gorgonian, 702 Dendronephthya sp., 703 Sinularia kavarattiensis, 704 Echinogorgia sassapo reticulata 705 and Anthrogorgia ochracea. 706 Of these sesquiterpenes, rumphellaoic acid A 1093 697 contains a unique skeleton, while shagenes A 1102 and B 1103 702 and ochracenoid A 1109 706 contain rarely reported skeletons. In the case of the ochracenoid A, absolute conguration was assigned by use of TDDFT calculated ECD data.
A South China Sea collection of Anthrogorgia caerulea yielded, in addition to two known avermectin macrolides, avermectin B 2a and 22,23-dihydroavermectin A 1a , 707 two new congeners B 1c 1111 and B 1e 1112. 708 All four MNPs exhibited moderate antifouling activities.
Further examples of diterpenoids, 1213 and 1214, 739 1215 and 1216, 740 cespitulones A 1217 and B 1218, 741 dihydrosarsolenone 1219, methyldihydrosarsolenoneate 1220, and sarsolilides B 1221 and C 1222, 742 have been isolated from so corals of the genera Cespitularia, Xenia, and Sarcophyton. Structure elucidation and determination of absolute conguration of 1219 and 1220 742 led the authors to propose a revision of the relative conguration at C-2 of the previously reported MNP sarsolenone (to that shown 1223). 743 Absolute conguration was assigned to 1219 and 1220 using TDDFT calculations of ECD data.
Sarcophyton ehrenbergi was the source of a number of prostaglandins including sarcoehrendins A-J 1224-1233. 744 In addition, three prostaglandins previously reported as synthetic compounds 1234-1236 were reported as natural products for the rst time. Relatively potent activity was observed towards phosphodiesterase-4, a target for CNS, inammatory and respiratory diseases. So corals also yielded a variety of steroids including pregnanes 1237 and 1238, 745 seco-sterols 1239-1244 746,747 and hydroxylated/polyhydroxylated sterols 1245-1265 747-753 from species of Schleronephthya, Subergorgia, Sarcophyton, Sinularia, Verrucella, Echinogorgia, and Leptogorgia. Noteworthy amongst these compounds, was the enhanced (synergistic) cytotoxicity observed for paclitaxel in the presence of punicinols A 1257 and B 1258 (Leptogorgia punicea) and that the two sterols were also able to inhibit A549 tumour cell growth in a clonogenic assay over a sustained period of ten days. 752 In addition to the mildly antiproliferative epoxyergosterols 1266-1271 (Anthopleura midori), 754 investigation of MNP chemistry of sea anemones and hydrozoa has identied two new cytolysins (3013 and 3375 Da) from the tentacles of the hydrozoan Olindias sambaquiensis 755 and PhcrTx1, a 32 amino acid residue acid-sensing ion channel inhibiting peptide from the sea anemone Phymanthus crucifer 756 that represents the rst example of a peptide containing an Inhibitor Cysteine Knot scaffold to be isolated from a cnidarian. Further study of the biological activities of previously reported anemone toxins has identied Av3 (Anemonia viridis) to show specicity towards arthropod voltage-gated sodium channels by binding to one of the transmembrane cles of the channel a-subunit, 757 while AdE-1 (Aiptasia diasphana) prolongs cardiomyocyte action potential duration while lowering peak amplitude via slowing inactivation of sodium channels and enhancing the transient K + current. 758 Mutation of the pore-forming toxin sticholysin I (Stichodactyla helianthus) residue tryptophan 111 to cysteine reduced the toxins affinity for membranes by an order of magnitude, 759 cysteine mutants of phenylalanine 15 or arginine 52 did not alter pore-forming activity but did protect the toxin from peroxynitrite oxidative damage, 760 while a third study, using monolayers of phosphatidycholine and sphingomyelin, determined that the toxin preferentially binds and penetrates membranes which have moderate enrichment in sphingomyelin and membrane uidity. 761 The rst total syntheses of alkaloids N-  773 to intricarene (Pseudopterogorgia kallos) 774 via a photochemical pathway has been demonstrated, 775 while a photochemical (E/Z) olen isomerisation was a critical step in the total synthesis of the natural product analog 17-deoxyprovidencin. 776 Clarication of the structure of the cladiellin diterpene sclerophytin F (Sclerophytum capitalis) 777 has been an ongoing issue, with Friedrich and Paquette in 2002 proposing the structure should be revised to be the (3S) diastereomer of sclerophyton A. 778 Synthesis of this proposed revised structure gave a product whose spectroscopic data differed from those reported for the natural product, suggesting further investigation is needed to resolve this issue. 779 A modular approach to a number of cladiellin MNPs using a gold-catalysed tandem reaction of 1,7-diynes has been reported. 780 Hippuristanol (Isis hippuris) 781 analogues have been evaluated as inhibitors of eukaryotic translation initiation, 782 cembranoid and ergosterol MNPs from Vietnamese cnidarians were found to exhibit selective in vitro activity against T. brucei, 783 a number of diterpenes (Sinularia maxima) were found to be modest to poor inhibitors of NF-kB transcriptional activation, 784 structurally simpler analogues of fuscol/eunicol exhibit comparable or better anti-inammatory activity in the mouse ear edema assay, 785 semi-synthetic oxygenated dolabellane diterpenes exhibit in vitro anti-HIV activity, 786 further semisynthetic examples of hydoxysterols were found to induce pregnane X receptor transactivation, 787 exibilide 788 exhibits in vivo anti-neuroinammatory activity in rats, 789 11-epi-sinulariolide acetate inhibits carcinoma cell migration and invasion by suppressing a number of phosphorylationdependent pathways, 790 13-acetoxysarcocrassolide induces apoptosis in carcinoma cells by activation of p38/JNK and suppression of PI3K/AKT pathways, 791 and cembranoids from Sarcophyton glaucum exhibit cytotoxicity towards a murine melanoma cell line. 792,793

Bryozoans
There were only three reports (containing 19 compounds) of new metabolites isolated from bryozoans in 2014, which is a large increase on 2013 when only one new metabolite was reported from this understudied phylum. The bromopyrrole alkaloids aspidostomides A-H 1272-1279 and aspidazide A 1280 were isolated from the Patagonian bryozoan Aspidostoma giganteum. Aspidostomide E 1276 exhibited moderate inhibition of the 786-O renal carcinoma cell line. Aspidazide A 1280 is a rare asymmetrical diacylazide and NOE NMR correlations and chemical transformations were utilised in determination of the structure. 794 A series of ceramides neritinaceramide A-E 1281-1285 were obtained from Bugula neritina and exhibited selective but weak activity against two HTCLs. 795 The same sample yielded several sterols 1286-1290. 796 Convolutamydine A 797 displayed signicant anti-inammatory activity both in vitro and in vivo. 798

Molluscs
The 23 new metabolites reported from molluscs is the average number reported per year over the past decade. 37-epi-Azaspiracid-1 1291 was isolated from contaminated raw shell-shthe epimer forms spontaneously via an equilibrium and formation, as expected, is accelerated by heating. 799 Implication of the latter in relationship to the cooking of shellsh is important as 37-epi-AZA1 was 5-fold more toxic towards Jurkat T lymphocyte cells in vitro than AZA1. Corresponding epimers of the related toxins AZA2 and AZA3 were also detected.
Specimens of the Mediterranean sacoglossan mollusc Thuridilla hopei afforded new nor-diterpene aldehydes 1292-1294 800 in addition to known congeners thuridillin A, B and C. 801 The mollusc feeds on the green alga Derbesia tenuissima, extracts of which only contain a known epoxylactone, 802 supporting the assumption that 1292-1294 are mollusc transformation products. Acetylenic alcohols 1295-1299 were isolated from both the Mediterranean dorid nudibranch Peltodoris atromaculata and one of the nudibranch's common dietary prey, the sponge Haliclona fulva. 803 In a similar manner, renieramycin-related alkaloids fennebricin A 1300 and B 1301 were isolated from both the nudibranch Jorunna funebris and the sponge Xestospongia sp. 804 The substructurally-related isoquinoline 1302 was only identied in the sponge extract. New diketopiperazines 1303-1306 were reported from Pleurobranchus areolatusrelated metabolites have been reported from the ascidian Didemnum sp., a suspected prey of P. areolatus. 805 Further investigation into the origin of tetrodotoxin in Pleurobranchaea maculata demonstrated that the mollusc can accumulate the toxin through its diet however there was no identiable tetrodotoxin source in the molluscs local environment. 806 Two studies of sea hares identied two moderately cytotoxic sesquiterpenes oculiferane 1307 and epi-obtusane 1308 (Aplysia oculifera) 807 and an anti-neuroinammatory diterpene dactyloditerpenol acetate 1309 (A. dactylomela). 808 6-Bromoisatin, typically isolated from muricid molluscs, is weakly antiproliferative towards HT29 cells, and enhances apoptosis in an in vivo colon cancer model. 809 Further structure-activity relationship studies of the cyclic peptide sanguinamide B (Hexabranchus sanguineaus) 810 have found that antiproliferative activity is dependent upon both the location of specic amino acids in the macrocycle and their conguration. 811,812 New synthetic auristatin analogues, being related to the original MNP dolastatin 10 (Dolabella auricularia), 813 bearing changes at the N-terminus showed pronounced antitumour activity. 814 The binding of three examples to tubulin was investigated by co-crystal X-ray studies, identifying an interesting structural feature whereby in the solid state the valine-dolaisoleucine fragment exists in the cis-conguration, whereas in solution it exists solely in the trans-conguration. In further studies, cholesterol and other simple sterols puried from mussels showed anti-aging and neuroprotective properties, 815 synthetic tambjamine analogues show enhanced tumour cell antiproliferative and chloride transport properties, 816 and lamellarin D (Lamellaria sp.) 817 induces senescence in cancer cells, in a process that includes the generation of intracellular ROS and requires the presence of topoisomerase I. 818 Biosynthesis of long chain polyunsaturated fatty acids in the scallop Chlamys nobilis was investigated, revealing the presence of a new elongase, that can elongate 20:4n-6 and 20:5n-3 to C22 and C24 acids, and a D 8 -desaturase. 819 A new example of an a4/7-conotoxin, a-BnIA 1310 was isolated from crude venom of the molluscivorous cone snail Conus bandanus. 820 Peptides with the same sequence, Mr1.1 and Bn1.1, were previously identied by PCR amplication of venom duct cDNA from molluscs C. marmoreus 821 and C. bandanus. 822 The peptide reversibly inhibited the human a7 nicotinic acetylcholine receptor (nAChR) and blocked nerve-evoked skeletal muscle contractions. Conus bandanus (Vietnam) was also the source of BnIIID 1311, a 15 residue M-1 family peptide containing six cysteines (disulde connectivity not determined) and three post-translational modications comprised of a bromotryptophan, g-carboxy glutamate and amidated aspartic acid residues. 823 An unusual a5/5 conotoxin AusIA 1312 was puried from the venom of C. australisboth synthetic globular (natural) and ribbon (different disulde linkages) congurations inhibited the a7 nAChR. 824 A short cyclic hexapeptide Vi804 1313 was isolated from crude venom of C. virgo and the solution structure of it and the D W3 synthetic analogue explored by NMR spectroscopy. 825 The high hydrophobicity of g-conotoxins make them difficult to synthesise by standard peptide synthesis techniques. A recent report describes the use of a Lys 4 solubilising C-terminus tag to enable the synthesis and Na V subtype selectivity of three previously reported C. consors g-conotoxins, g-CnVIB, g-CnVIC and g-CnVID. 826 Further studies have reported on the structure and activity of dicarba analogues of a-RgIA, 827 the inuence of disulde connectivity on structure and bioactivity of a-TxIA, 828 the inuence of acetylcholine to affect the binding of a-MII at nAChRs, 829 the neuronal target selectivity of the Conus textile T-superfamily peptide TxVC, 830 and the Ca 2+ -activated K + (BK) channel selectivity of the unusual M superfamily conotoxin Vt3.1. 831 11 Tunicates (ascidians)  838 Noteworthy was the isolation, structure elucidation and synthesis of a rare example of a modied nucleoside bearing a 5 0 -thiomethyl substituent 1333. 839 The same study also led to the reassignment of structure of a known sponge metabolite hamiguanosinol 840 from the enol tautomer to the guanosine keto tautomer 1334. The tanjungides A 1335 and B 1336, 841 novel dibromoindole enamide alkaloids isolated from Diazona cf. formosa exhibit potent cytotoxicity (<1-2 mM) towards a panel of HTCLs. An eleven-step linear synthesis utilising Buchwald vinyliodide amidation and peptide synthesis established the absolute conguration of the alkaloids.
Over the years a number of cyclic ribosomal peptides, now known as the cyanobactins, have been reported from ascidians, typically of the genus Lissoclinum. The true producers of these natural products are photosynthetic symbiotic cyanobacteria of the genus Prochloron. The apparent random distribution of cyanobactins isolated from ascidians has now been credited to host phylogeny, with genetic analysis revealing that 'Lissoclinum patella' falls into three phylogenetic groups that in turn may contain further cryptic species. 842 The implications that local extinctions of such cryptic species may reduce marine natural product diversity is indeed food-for-thought in the context of climate change. A very interesting demonstration of the use of the biosynthetic machinery of cyanobactin production was recently reported, whereby engineered enzymes of the patellamide pathway, in combination with enzymes from other cyanobactin-related pathways, were used for the in vitro production of a small library of cyclic peptides. 843 This proof of principle allowed for the preparation of 1-2 mg of each peptide. Total synthesis has led to revision of the original structure proposed for didemnaketal B (Didemnum sp.) 844 to 1337, requiring stereochemical inversion of the C-10-C-20 spiroacetal domain of the MNP. 845 Such a revision may be of relevance to the ongoing revision of the (stereo)structure of didemnaketal A. 846 Syntheses of the reported structures of polycitorols A and B (unidentied ascidian) 847,848 and (+)-didemniserinolipid C (Didemnum sp.) 849,850 suggest that the structures of all three MNPs require revision. Total synthesis has also led to revision of the structure of mandelalide A (Lissoclinum sp.) 851 to 1338 852 and conrmation of the revised structure of haouamine B (Aplidium haouarianum), [853][854][855] while the structures of distomadines A and B (Pseudodistoma aureum) 856,857 and synoxazolidinones A and B (Synoicum pulmonaria) 858,859 have been conrmed by synthesis. Synoxazolidinones A and C and S. pulmonaria co-metabolites pulmonarins A and B exhibited variable levels of anti-biofouling activity against a panel of test organisms, with synoxazolidinone C being particularly potent as both a growth and adhesion inhibitor. 860 The effects of ascidian extracts on the estrogen receptor-negative breast cancer cell line MDA-MB-231 were investigated by content-rich screening, leading to the identication of eusynstyelamide B (Didemnum candidum, originally isolated from Eusynstyela latericus 861 ) as a moderate cytotoxin (IC 50 5 mM) causing cell cycle arrest in G2/M and inducing apoptosis. 862 The potently cytotoxic macrolide iejimalide C (Eudistoma cf. rigida) 863 joins congeners iejimalides A and B as being identied as an inhibitor of the vacuolar-type ATP-driven proton pump (H + -ATPase). 864 Aer 24 h treatment, cells also exhibited actin aggregates, but as the MNP does not inhibit actin polymerisation in vitro, it was concluded that actin activity was a consequence of disruption of pH homeostasis. Preliminary data suggesting that trabectedin (Et 743) exhibits anti-angiogenic activity towards breast cancer cell lines has been reported. 865
A number of new saponins were identied in extracts of the Australian sea cucumber Holothuria lessoni using solely LC-MS/ MS metabolomic techniques. 876 In a conceptually similar manner, the chemical diversity of saponins present in different organs of the starsh Asterias rubens was investigated by combinations of MALDI-TOF and LC-MS(/MS) techniques. 877 The latter study concluded that different organs are characterised by different saponin mixtures and inter-specimen variability exists suggesting inuence of sex and/or collecting season on saponin prole. A short octapeptide echinometrin 1373 (sea urchin Echinometra lucunter) was found to exhibit ability to degranulate mast cells leading to an inammatory reaction. 878 The sequence of the peptide is an internal fragment of vitellogenin, a nutrient protein present in sea urchin gametogenic cells, suggesting the possibility that echinometrin is a cryptide. 879 The rst synthesis of a pyranonaphthazarin pigment isolated from the sea urchin Echinothrix diadema 880 has been reported. 881 Further studies using puried pentahydroxynaphthoquinone echinochrome A 882,883 have identied it to enhance mitochondrial biogenesis, 884 to protect cardiomyocyte mitochondria from the effects of cardiotoxic drugs 885 and to inhibit acetylcholinesterase in an irreversible and uncompetitive mode. 886 The mechanisms of antioxidant reactivity of the structurally related echinamines A and B 887 has been investigated using DFT methods. 888 The benzochromenone comaparvin (Comanthus bennetti) 889 exhibits anti-inammatory activity in the carrageenan-induced rat model. 890 A simple synthesis of ovothiol A 891 from L-hisidine has been reported, 892 the role of a non-heme iron oxidase enzyme in the biosynthesis of ovothiol A investigated, 893 NMR-pH titrations used to investigate the microspeciation of the amino acid, 894 and the antiproliferative activity towards Hep-G2 cell line via an autophagy mechanism identi-ed. 895 Sterols and fatty acids from the spiny sea-star Marthasterias glacialis exhibit both anti-inammatory 896 and cell cycle arrest properties, 897 while sterols from an urchin and a starsh exhibit selective antiparasitic activity towards T. brucei. 783 Investigation of the structure and bioactivity of chimeric analogues of the starsh cardiac-stomach relaxing SALMFamide neuropeptides S1 and S2 has highlighted the importance of the C-terminus for bioactivity and that the N-terminus confers structural stability. 898 Treatment of pancreatic cancer in vivo in athymic mice using a combination of frondoside A (Cucumaria frondosa) 899 and the nucleoside anticancer agent gemcitabine was signicantly more effective than with either drug alone. 900 The cytotoxic and apoptotic-inducing abilities of steroids from the cold water starsh Ctenodiscus crispatus 901 and the triterpene glycosides, including echinoside A, from Holothuria scabra and Cucumaria frondosa have been reported. 902 Echinoside A and related saponin holothurin A inhibit dietary fat absorption and decrease the adipose tissue accumulation in mice, 903 and typicosides (Actinocucumis typica) 904 exhibit a strong immunostimulatory effect on mouse macrophages with marked increase in lysosomal activity and ROS formation. 905 Complete NMR assignments have been reported for four pentaoside asterosaponins: thornasteroside A, versicoside A, anasteroside B and asteronylpentaglycoside sulfate (Asterias amurensis). 906

Mangroves
In addition to a series of mildly antioxidant glycosides, marinoids F-M 1374-1381, reported from the whole fruits of the mangrove Avicennia marina, 907 916 and 1423 917 (X. rumphii and X. granatum) were also isolated from mangroves or their associates. The absolute conguration of the unusual spiro-secoabietane decandrinin 1382 was secured via analysis of experimental and calculated ECD and ORD chiroptical data 909 while of the limonoids reported, the structures of xylorumphiin G 1398, 912 xylogranatopyridine A 1402, 913 granatumin L 1409 915 and granatumin Y 1422 916 were established by single crystal Xray diffraction studies.
Further investigation of previously reported mangrove MNPs has identied avicequinone C (Avicennia marina) 918 as a 5a-reductase-type 1 inhibitor, 919 catunaregin (Micromelum falcatum) 920 as an angiogenesis inhibitor, 921 the cardiac glycoside neriifolin (Cerbera manghas) 922 as the acaricidal component of Panonychus citri, 923 and that leaf extracts and puried components of Avicennia marina exhibit antibacterial activity with relevance to urinary tract infections. 924 Finally, a new biomimetic synthesis 925 and a structure-activity relationship study of the protein tyrosine phosphatase 1B inhibiting properties 926 of the unusual dimeric alkylbutenolide paracaseolide A (Sonneratia paracaseolaris) 927 have been reported.

Miscellaneous
Two studies of sea grass from the Egyptian Red Sea led to the identication of avone xyloside 1424 (Thalassia hemprichii) 928 and dihydrochalcone diglycoside 1425 (Thalassodendron ciliatum) 929 as antimicrobial and anti-inuenza A virus MNPs respectively. Thelepamide 1426 is an unusual ketide-amino acid isolated from the annelid worm Thelepus crispus. 930 The relative conguration of the alkaloid was determined by a combination of heteronuclear J-based congurational analysis and GIAO calculated chemical shis and DP4 probability analysis. Mild cytotoxicity towards a leukemia cell line was also observed. The arenicins, antimicrobial peptides produced by the polychaete worm Arenicola marina, 931 are constitutively expressed in a range of tissues in the organism suggestive that the peptides play a front-line role in defense against infections. 932 Cypridina luciferyl sulfate 1427 appears to be a more stable storage form of cypridina luciferin, the luminescence precursor of the ostracod Cypridina (Vargula) hilgendori. 933 The luciferin could be converted to luciferyl sulfate by action of crude extract of the organism, presumably containing a sulfotransferase, in the presence of 3 0 -phosphoadenosine 5 0 -phosphosulfate (PAPS, a sulfate donor), while the reverse reaction took place in the presence of adenosine 3 0 ,5 0 -diphosphate, a sulfate acceptor.
Extracts of ovary tissue from Takifugu pardalis yielded a new tetrodotoxin analogue, 6-deoxyTTX 1428. 934 The potent voltagegated sodium channel blocker was also detected by LC-MSMS in other marine animals including snail and octopus. Alanine scanning of the hagsh (Myxine glutinosa) 935 12-amino acid residue antimicrobial peptide myxinidin has identied a number of critical residues for the observed biological activities and that judicious substitution with arginine led to the identication of more potent analogues. 936 The 33-amino-acid residue peptide pardaxin (atsh Pardachirus marmoratus) exhibited in vivo activity towards MRSA dermal infection and enhanced wound healing. 937 A range of arsenic-containing lipids have been synthesised, providing valuable reference standards for future studies directed towards understanding the uptake, biotransformation and toxicity of these unusual MNPs. 938

Conclusion
In a recent review the number of new MNPs reported from a range of countries was analysed based on the location of the principal author for each publication. 3 What is not apparent from this analysis is the location of the collecting sites for the organisms. This can be problematical as shown in an extreme example: one prominent MNP chemist based in the Mid-West of the USA, thousands of km from any coastal resource, collected widely (see Fig. 1).
With the biogeographic aspect of the MarinLit database 19 now widely available an alternative perspective is available for viewing the collection data. It is now possible to search by region showing who is collecting where and hence the interests that MNP chemists have in the various regions of the globe. The 50 years of collecting effort globally is depicted in Fig. 2. The red stars in Fig. 2 represent the collecting sites, described in $9000 papers, from which $25 700 new compounds have been obtained. The actual number of unique collection sites is less than the number of papers as multiple collections have oen been made at the same sites. Fig. 2 displays those areas that have had high collecting pressures and those where there has been, for whatever reason, lower collecting pressures. An analysis has been made of the collecting effort globally by semi-decade since 1965. For convenience the globe was divided up into 22 regions or countries (see Table 1) and the published papers for each region compiled. Totals for papers (citing collection sites) describing new compounds from 1965 are also listed in Table 1 while Fig. 3 shows the incidence of discovery of new compounds in each region or country by semi-decade since 1965. It is worth noting that since 1965-280 papers contained no biogeographic data, not even a country or an ocean, to describe the origin of new compounds being reported. Fortunately, recent years have seen the increasing use of exact coordinates. The Japanese region, including Okinawa, has been the most productive region with 3877 compounds described. This was followed closely by China with 2915 compounds from the mainland with a further 525 compounds to be included if the regions of the South China Sea and Yellow Sea are also considered. But what is remarkable here is the rapid emergence of MNP chemistry using Chinese-based collections as seen in Fig. 3. Other regions where there have been extensive collections are in the Mediterranean (2358) (which includes the Mediterranean coasts of Spain and France, Italy, the islands of the Mediterranean, Greece, Turkey, Israel, Egypt, the Arabian Peninsula, Black Sea and the Mediterranean African coastlines), Australia (1854), the North (1539) and South (1548) Pacic islands and atolls, and the Gulf of Mexico, Caribbean Sea and islands (1524). Other regions that have been well explored include Taiwan (1350) and Maritime SE-Asia and Papua-New Guinea (1340).
The graph shows very clearly those countries that were most closely studied in the early years. Compounds of Japanese origin were prominent from the 1960s, but the search for MNPs was quickly taken up with compounds from the Mediterranean, North Pacic and North American regions appearing in the early 1970s, followed by Australian and Maritime SE-Asian compounds later in that decade. Prospecting activity in other parts of Asia was relatively slow to start and it was not really until the 1990s that compounds of Mainland SE-Asian, South Korean, Taiwan and Chinese origin started to appear. The output from these regions has rapidly accelerated since, particularly so in the case of compounds of Chinese origin. Fig. 2 and 3, taken in combination, provide a snapshot of the past efforts in marine natural products as well as current endeavours and highlights those areas of the globe that are currently under-explored.