In our continuing research on the cytotoxic constituents of Taiwanese gorgonian corals, the EtOAc extracts of Briareum excavatum (Nutting) and Junceella fragilis (Ridley) were investigated, repectively. Twenty-four compounds, excavatolides D (1), F-K (2-7), M (8), U-Y (9-13);briaexcavatolides D (14), K (15), L (16), O(17), P (18); excavatolides B (19), C (20), E (21); briaexcavatolides B (22), C (23), and (1S*,2S*,5Z,7S*,8S*,9S*,10S*11R*,12R*,13Z,17R*)-2,12-diacetoxy-8,17-epoxy-9 hydroxy briara-5,13-dien-18-one (24) were isolated from B. excavatum. Three compounds, junceellolides E (25), F (26), and umbraculolide A (27) were isolated from J. fragilis. Among them, compounds 1-18, 25, and 26 are new products. All metabolites 1-27 are briarane-type compounds. The structures of metabolites 1-27 were determined by physical (mp, optical rotation) and spectral (UV, IR, MS, HRMS, 1D, and 2D NMR) data analyses and chemical methods and by comparison with the related physical and spectral data from other known briarane-type compounds. The structures, including the relative configurations of metabolites 1, 9, 15, 17, 18 and 25 were further confirmed by X-ray single-crystal analyses. Furthermore, the S-trans diene system in 1; the propionyloxy group at the C-4£] position of 13; the hydroxy groups at the C-8£]and C-17£\positions of 15 and 16; the independent functional groups at the C-2£],C-3£] and C-4£\ positions of 17 and 18, and the boat conformation in the six-membered ring of 25 and 26, all are the first observations in the briarane-type natural products. The cytotoxicity of the isolates against the P-388(mouse lymphocytic leukemia), KB (hum nasopharyngeal carcinoma), A549 (human lung adenocarcinoma), and HT-29 (human colon carcinoma) cancer cell lines were studied. In the cytotoxicity testing, 1, 6-8, 10, 16, 18, 20-22, and 24 show significant cytotoxicity against the grouth of P-388 cells; 7, 8, 20-22, and 24 show significant cytotoxicity toward KB cells; 7, 8, 20, and 21 show significant cytotoxicity toward A549 cells; 1, 7, 8, 18, 20, and 21 show significant cytotoxicity toward HT-29 cells.

In order to discover and develop new drug from soft corals and gorgonian corals of Taiwan, we have searched the bioactive metabolites from the organic extracts of three soft corals Nephthea chabrolii, Sinularia manaarensis, Sinularia leptoclados, and one Gorgonian of Briareum genus. This study had led to the isolation of fifty-five natural products (1¡V55), including seventeen new meroditerpenoid-related metabolites (1¡V7 and 9¡V18), two new C18 terpenoid¡Vrelated carboxylic acids (20 and 21), three new sesquiterpenoidalnatural products (22¡V24), and two new steroids (26 and 27), along with three known compounds (8, 19, and 25) from N. chabrolii; nine new cembrane-type diterpenoids (28¡V36), along with two known cembranolides 37 and 38 from S.manaarensis; five new sesquiterpenoids (39¡V43), one steroid (47) isolated for the first time from natural sources and three known metabolites (44¡V46) from S. leptoclados; and three new briarane-type derivatives (48¡V50) and five known briarane-type compounds (51¡V55) from Briareum sp. The structures of metabolites 1¡V55, including their stereochemistries have been established by detailed spectroscopic analyses, particularly mass, 2D NMR (1H¡V1H COSY, HMQC, HMBC, and NOESY) spectroscopy and by comparison with the related physical and spectral data form other known compounds. The relative configuration of metabolite 39 was further confirmed by X-ray single-crystal analysis. Furthermore, the biosyntheses of meroditerpenoids 1, 4, 11, and 20 were proposed. To the best of our knowledge, the incorporation of a methyl group of the related meroditerpene to form a naphthoquinone as discovered herein for the first time. In above metabolites, two compounds (2 and 10) were found to exhibit significant inhibition against the growth of MCF 7, NCI-H460, and SF-268 tumor cells at 20 £gg/mL. Also, compound 2 exhibited significant cytotoxicity against the growth of MDA¡VMB¡V231cancer cell line, and moderate to weak cytotoxicity against Hep G2 and A-549 cancer cell lines and metabolite 10 exhibited moderate to weak cytotoxicity toward MDA¡VMB¡V231, Hep G2 and A-549 cancer cell lines. Furthermore, Two cembranolides (34 and 35) exhibited moderate cytotoxicity against Hepa59T/VGH, KB, Hela, and Med cell lines.

The sexual reproduction of four gorgonian corals, Ellisella robusta, Subergorgia suberosa, Subergorgia mollis and Bebryce indica at Wanlitong, a non-upwelling area, and Talauko, an upwelling area, in south Taiwan were compared in order to understand their reproductive strategies. Four species were gonochoric. E. robusta, S. suberosa and S. mollis were broadcasting spawners with annual reproductive cycle. The reproductive mode of B. indica was not certain and it may reproduce several times within one year. The mean diameter of mature oocytes of E. robusta, S. suberosa, S. mollis and B. indica was 360, 322, 461, and 312 £gm, respectively. Their fecundity was 3.2, 1.4, 1.1, and 2.0 oocytes/polyp, respectively. Corals with longer oogenesis duration produce larger mature oocytes. The three broadcasting species spawned in September, October and November after the seasonal disturbances. It may be advantageous for the survival of their offspring. The reproductive traits of E. robusta, S. suberosa, and B. indica were similar between Wanlitong and Talauko populations. It suggests that reproductive traits of theses species may be not influenced by upwelling.

This thesis examined the reproductive biology of the poorly-known deep-water gorgonian Acanella arbuscula from the Northwest Atlantic. Colonies were collected from The Gully in 2007 and 2010 between 914 and 1860 m depth, and the Flemish Cap in 2009 between 671 and 1264 m. Mean polyp fecundity was relatively high for both females and males, and the large oocyte size suggests that A. arbuscula produces lecithotrophic larvae. This species may have overlapping periodic or seasonal cycles of gametogenesis, and the absence of planulae suggests that A. arbuscula is a broadcast spawner. No spatial variation in the reproductive characteristics of this species was found, suggesting that environmental conditions are similar between the two sites. Female polyp fecundity decreased with increasing depth, which may be due to the high cost of producing oocytes versus sperm. The relatively high mean polyp fecundity, probable lecithotrophic larval development, and broadcast spawning may allow for the wide dispersal and settlement of A. arbuscula across the North Atlantic.

Abstract Several sesquiterpenes isolated from a Formosan gorgonian coral Isis hippuris have shown significant cytotoxic activity against various cancer cell lines. In order to search other active components, we have investigated the chemical constituent of I. hippuris from a Green Island specimen. In the cytotoxcity assay, the EtOAc extract showed potent cytotoxic response toward P-388, A549 and HT-29 cancer cell lines. Thus, the investigation on the chemical content of this extract was carried out. This study finally led to the isolation of twelve steroids(1-12). Nine metabolites, hippuristerone G (1)¡Bhippuristerone H (2), hippuristerone I (3)¡Bhippuristerone J (4)¡Bhippuristerol E (5)¡B2a,3a-diacetoxy-24-methyl- 11b,18; 18,20b; 22,25-triepoxy-5a-furostane (7)¡B2a,3a-diacetoxy-11b- hydroxy-24-methyl-22,25-epoxy-5a-furostan-18,20b-lactone (10)¡B3a- acetoxy-11b,18a-dihydroxy-24-methyl-18,20b; 22,25-diepoxy-5a- furostane (11) and 2a,3a-diacetoxy-11b,18a-dihydroxy-24-methyl-18, 20b; 22,25-diepoxy-5a-furostane (12) are new compounds, whereas hippuristerone A (6), 3a-acetoxy-24-methyl-11b,18; 18,20b; 22,25- triepoxy-5a-furostane (7) and 3a-acetoxy-11b-hydroxy-24-methyl-22,25- epoxy-5a-furostan-18,20b-lactone (9) are known compounds. The structures of 1-12 were elucidated by spectroscopic evidences (IR, MS, 1D NMR, 2D NMR) and chemical method. The stereochemistries of compounds 7 and 9 were further confirmed by single-crystal X-ray diffraction analyses. Cytotoxicity test revealed that hippuristerone I (3) exhibited moderate inhibition toward NCI cancer cell line.

This dissertation mainly presented the investigation of natural products from two different Formosan gongonian soft corals, Briareum violacea and Junceella fragilis. Their extracts were examined by intensive chromatographic methods. Eighty-four compounds including fifty new briaranes were isolated, and parts of their biological activities were studied. Natural products investigation of the Taiwanese gorgonian octocoral Junceella fragilis found fifteen new briarane-type diterpenes, namely frajunolides E-S (1-15), ninteen known briaranes, eg. junceellin, junceellolides A-E¡Bjunceellolide K, 11£\,20£\-epoxy-4-deacetoxyjuncellolide D, umbraculolide A, junceellonoid A, juncin P, juncins Y-ZI, frajunolides A-D and praelolide, and a sterol, ergosterol peroxide. Soft coral B. violacea (Quoy and Gaimard) of Taiwan has isolated thirty-five new briarane-type diterpenoids, briaviolides A-Z (16-41) and viobrianolides A-I (42-50), in addition to fourteen known diterpene lactones, stylatulide lactone, excavatolide A, 9-deacetylstylatulide lactone, 4£]-acetoxy-9-deacetylstylatulide lactone¡B Brianthein Z¡B, renillafoulin A, braexcavatolide E, braexcavatolide I, minabein-4, minabein-6, milolide K, solenolide A, and solenolides D-E. Structures of all above compounds were examined by physical and spectroscopic analyses including mass, IR, UV spectra, optical rotation and 1D, 2D NMR, as well as in comparison with the published data. Moreover, the structures of compounds 16, 32 and prarlolide were further confirmed by single X-ray crystallographic analysis. The stereochemistry of these compounds was investigated by NOESY method, and the configuration of compounds 32-38 were determined by Circular Dichroism spectroscopic analysis. Cytotoixcity and in vitro anti-inflammatory activities were studied by Dr. Kuo, Yao-Haur and Dr. Hwang, Tsong-Long, respectively. Junceellin, junceellolide B and juncin ZI exhibited weak cytotoxicity against Hep2 (Human laryngeal carcinoma), Doay (Human medulloblastoma), WiDr (Human colon adenocarcinoma), Hela (Human cervical epitheloid carcinoma). The in vitro anti-inflammatory activities of 1-7 were evaluated for inhibition of elastase release and for the generation of superoxide anion, as tested on human neutrophils. Compounds 1 and 6 exhibited weak inhibition of elastase release and superoxide anion at 10 £gg/mL. The biological activity analysis of compounds 16-50 are in progress.

<p> Primnoa resedaeformis is a deep-sea gorgonian coral with worldwide distribution and a lifespan of at least several hundred years. Recent work has suggested that it may be possible to obtain extended, high-resolution records of ambient oceanographic conditions from Primnoa skeletons. This thesis focuses on specimens recently collected live from the Northeast Channel, SW of Nova Scotia, from depths of 300-500m. </p> <p> Skeletal microstructure was examined as a prerequisite to geochemical sampling. Skeletons exhibit periodic growth at three distinct scales. Concentric annual rings throughout the skeleton, and sub-annual laminae in the horny axis, measure 200 +1-100 microns and 15 +1-10 microns, respectively. Fine-scale striae in the outer calcite cortex measure 1.5 +1-2 microns. The dark, gorgonin-rich portion of annual rings in the horny axis forms in winter, when currents in the NE Channel are most energetic. Growth in these animals is apparently tied to the passage of currents at seasonal, lunar and tidal frequencies. Annual ring widths in the horny axis could not be successfully cross-dated, however, a prominent dark ring that appears to have been formed in 1976 is present in several of the colonies examined. Prominent dark rings may serve as useful benchmarks in sclerochronology. </p> <p> Mg/Ca and Sr/Ca were measured by laser ablation ICP-MS in the predominantly calcite axial cortex. Across a 1.5°C gradient, Mg/Ca is positively related to temperature. Sr/Ca also increases with temperature, but this may be explained by the influence of Mg/Ca on Sr partitioning, rather than temperature. Near annual-resolution timeseries profiles of Mg/Ca are consistent within and among colonies having different growth rates. Conversion of Mg/Ca profiles to temperatures using a provisional calibration [Mg/Ca (mmollmol) = 4.88(+/-1.09) T (°C) + 70.92 (+/-6.79)] yields a range of values and trends that are consistent with the observational data. Mg/Ca in Primnoa, therefore, is a viable means of monitoring bottom-water temperatures. The North Atlantic Oscillation (NAO) is responsible for a significant component of inter-annual temperature variability in the Scotia-Maine region. Mg/Ca records from older corals could therefore provide extended proxy records of the NAO. </p> / Thesis / Master of Science (MSc)

Coral diseases have become more prevalent and destructive over the past 20 years, possibly due to an increase in stressful environmental factors that may weaken corals defenses against disease. Aspergillosis is a disease caused by the fungus Aspergillus sydowii, which apparently infects only two species of gorgonian corals in the Caribbean Ocean (Gorgonia ventalina and G. flabellum). We hypothesized that the differential resistance to infection is caused by differences in chemical defenses among gorgonians. Freeze-dried gorgonian powders and extracts deterred fungal growth, but potencies varied among gorgonian species and among fungi. Extracts and powders generated from G. ventalina all strongly inhibited fungal growth. Since G. ventalina was predicted to have weak antifungal chemical defenses compared to gorgonians not known to suffer from aspergillosis, we concluded that gorgonian susceptibility to fungal infection is determined by factors other than, or in addition to, chemical defenses. In order to investigate specific gorgonian antifungal strategies, we attempted to use bioassay-guided fractionation to isolate antifungal compounds from four gorgonians: Gorgonia ventalina, Briareum asbestinum, Eunicea succinea, and Pseudopterogorgia americana. We succeeded in isolating two antifungal compounds, diastereomers of 9,11-seco-24-hydroxydinosterol, from the gorgonian Pseudopterogorgia americana. This compound was previously identified by other groups, but this study is the first to establish its antifungal activity. At natural concentration, one diastereomer of 9,11-seco-24-hydroxydinosterol inhibited the growth of three different fungi, suggesting that at least this diastereomer may possess broad-spectrum antifungal activity. The results from our survey of gorgonian chemical defenses indicate that susceptibility to aspergillosis cannot be explained by chemical growth inhibition alone. Further areas of investigation include induction of gorgonian chemical defenses, examination of growth-inhibiting mechanisms of antifungal metabolites, and identification of non-chemical factors affecting gorgonians vulnerability to fungal infection.

This research focuses on diterpenoids from Taiwanese soft corals Xenia umbellata Lamarck, Junceella juncea Pallas and Junceella fragilis Ridley. Twelve diterpenoids in addition to one sesequiterpenoid were isolated. Our investigation of the soft coral X. umbellata Lamarck afforded five natural products, including two new xenicane diterpenes, xenibelatols A-B (1-2), together with two known xenicane diterpenes, 7,8-oxido- isoxeniolide (3), 9-hydroxyxeniolide-F (4), and a cadinene sesequiterpene, xenitorin A (5). Chemical investigation of the gorgonian J. juncea Pallas, has resulted in isolation of a new briarane diterpene, juncenolide H (6). Continuing our investigation of the gorgonian J. fragilis Ridley, we isolated seven briarane diterpenes, including four new briaranes, flajunolides A-D (7-10), along with three known briaranes, junceellolide E (11), umbraculolide A (12), 11This research focuses on diterpenoids from Taiwanese soft corals Xenia umbellata Lamarck, Junceella juncea Pallas and Junceella fragilis Ridley. Twelve diterpenoids in addition to one sesequiterpenoid were isolated. Our investigation of the soft coral X. umbellata Lamarck afforded five natural products, including two new xenicane diterpenes, xenibelatols A-B (1-2), together with two known xenicane diterpenes, 7,8-oxido- isoxeniolide (3), 9-hydroxyxeniolide-F (4), and a cadinene sesequiterpene, xenitorin A (5). Chemical investigation of the gorgonian J. juncea Pallas, has resulted in isolation of a new briarane diterpene, juncenolide H (6). Continuing our investigation of the gorgonian J. fragilis Ridley, we isolated seven briarane diterpenes, including four new briaranes, flajunolides A-D (7-10), along with three known briaranes, junceellolide E (11), umbraculolide A (12),11£\, 20£\-epoxy-4-deacetoxy junceellolide D (13). The new compounds 1,2 and 6-10 possess xenicane-type and briarane-type skeletons respectively. The structures of new compounds were determined by 1D-, 2D-NMR spectroscopic analysis and physical methods such as optical rotation, UV, IR, mass spectrum, as well as comparison with the spectroscopic data reported for related compounds. Compounds 1 and 2 are geometric isomers of compounds 3 and 4. The only difference between them resides in the side chain. The geometry of the side chain influenced the relative spatial proximity of H-12, H-13, H-14 to the carbonyl at C-3, and consequently the extent to which these protons are subjected to the anisotropic effects of the carbonyl. Compounds 6-10 have acetyl groups at C-2, C-9, C-12, C-14 positions. Because of structural difference appears in briarane skeleton, they showed different chemical shifts in specific positions. Biological activity test¡Arevealed that compound 5 exhibited moderate cytotoxic activity against KB and WiDr cancer cell lines with ED50 values at 5.9 and 9.9 £gg/ml respectively.

Abstract The n-hexane extract of Formosan gorgonian coral Isis hippuris (Isididae) collected from Green Island was found to exhibit significant cytotoxicity against P-388, A-549 and HT-29 cell lines. Cytotocicity-guided fractionation of this extract led to isolation of a known subersenone (1), five new suberosanane analogues, suberosanone (2), suberosenol A (3), suberosenol B (4), suberosenal A acetate (5) and suberosenol B acetate (6), two sesquiterpenes of novel skeletons, isishippuric acid A (7) and B (8). Three known compounds, subergorgic acid (9). 3-hexadecyl-5-hydroxy- 5-methyl-5-hydrofuran-2-one (10) and prenga-1,4-diene-3,20-dione (11) were also isolated along with the novel compound. Compounds 2-6 and 8 showed potent cytotoxicities against P-388, A-549 and HT-29 cells.