Abstract 24£i-methylcholestane-3£],5£\,6£],25-tetrol-25-monoacetate (1), 24£i-methyl cholestane-3£],5£\,6£],25-tetrol (2), 24£i-methylcholestane-1£],3£],5£\,6£],25-pentol -25-monoacetate (3), (20S)-21-decanoxyl-24£i-methylcholestane-3£],5£\,6£],24 -tetrol (4), (20S)-21,24-diacetaoxy-24£i-methylcholestane-3£],5£\,6£]-triol (5) were isolated from the soft coral Sarcopyton glaucum which were collected from the Green island. The structures of 1~5 were established by spectroscopic data , compound 1~3 have been reported previously, compound 4 and 5 are new metabolites. The cytotoxicity of compounds 1~5 against A-549, HT-29 and P388 cancer cell lines was studied. Only compound 1 showed significant cytotoxicity against the growth of HT-29 cells line. Compound 2~5 were found to be inactive against the above three cancer cells lines.

Chromatographic purification of a methylene chloride extract of Formosan soft coral Sarcophyton crassocaule (collected in Green island) led to the isolation of two new (1S*, 3R*, 4R*, 7E, 11E, 14R*)-3, 4-epoxycembra-7, 11, 15-trien-1, 14-olide (1) and (1R*, 3E, 7E,11R*, 12S*, 14R*)-11, 12-epoxycembra-3, 7, 15-trien-1, 14-olide (2) isomeric diterpenoids , a known cyclic peroxide diterpenoid (1R*, 2S*, 3E, 7S*, 8R*, 11S*, 12Z)-8, 11-epidioxy-7-acetoxycembra-3, 12, 15-trien-1, 2-olide (3), as well as two known (24S)-24-methylcholestane-3b, 5a, 6b-triol (4) and 24x-methyl-cholestane -3b, 5a, 6b, 25-tetrol 25-monoacetate (5) steroids. Chromatographic fractionation of a methylene chloride extract of two Formosan soft corals Sarcophyton elegans (collected in Green island) led to the isolation of a known 24x-methylcholestane -3b, 5a, 6b, 25-tetrol 25-monoacetate (5) steroid, the methylene chloride extract of Sarcophyton trocheliophorum, on the other hand, afforded a known diterpenoid, (+)-isosarcophine (6). Purification of a methylene chloride extract of Octocorallia soft coral (unidentified) led to the isolation of two known steroids, cholesterol (7) and (22E, 24S)-24-methylcholesta-5, 22-dien-3b-ol (8). Compounds 1-7 exhibited cytotoxicity against P388 cancer cell line. Compounds 2 and 5 were active against HT-29 cancer cell line.

In order to search for novel bioactive compounds, we have investigated the secondary metabolites of the soft corals Nephthea erecta, Lobophytum durum, and Sarcophyton ehrenbergi. Chemical examinations on the organic extracts of N. erecta, have resulted in the isolation of six new sesquiterpenoids (1-6), a new calamenene-type sesquiterpene with a mercaptan group, erectathiol (7), a new tri-nor-eudesmadienone (8), two known sesquiterpenoids (9 and 10), one novel seco-germacrane sesquiterpene (11), three unexpected artificial 19-oxygenated steroids (12-14), as well as twelve new polyhydroxylated steroids (15-26). Furthermore, twelve new cembranolids, durumolides A-L (27-38), three unprecedented hemiketal cembranoids, durumhemiketalolides A-C (39-41), six previously described cembranolids (42-47), together with one known metabolite, peridinin (48), were isolated from the acetone extracts of L. durum. Chemical investigation of S. ehrenbergi, has led to the isolation of a known ceramide (49), two new cerebrosides, sarcoehrenosides A (50) and B (51), along with three previously characterized cerebrosides (52-54). The structures of the isolated metabolites were elucidated through extensive spectroscopic analyses, while the relative stereochemistry of 10 and 44 were further confirmed by X-ray diffraction analyses. Moreover, the absolute configurations of 24, 25, 29, 34, 38, and 43-45 were established by application of modified Mosher¡¦s method. The cytotoxicities, antibacterial activities, anti-inflammatory effects, and inhibition assay of HCMV (human cytomegalovirus) endonuclease activities of these isolated metabolites were evaluated in vitro.

Thesis (Ph. D.)--University of Hawaii at Manoa, 1995. / Includes bibliographical references (leaves 124-127). / Microfiche. / xi, 127 leaves, bound ill. 29 cm

Marine organisms have attracted much attention as potential source for drugs over recent years. Soft corals have yielded many bioactive metabolites. Some of them have been examined for their pharmacological properties. For the process of drug discovery, we have examined bioactive metabolites from the organic extracts of two soft corals Sinularia gibberosa and Sarcophyton sp. collected off Formosan coast. This study had led to the isolation of forty-two natural products (1¡V42), including one new £]-caryophyllene-type sesquiterpenoid (1), four new xeniaphyllane-type norditerpenoids (2, 14, 16, and 17), fourteen new xeniaphyllane-type diterpenoids (3¡V13 and 18¡V20), one novel nor-humulene (15), seven new xeniaphyllane-type diterpenoids (21¡V26) with cyclic peroxyhemiketal (3,6-dihydro-1,2-dioxin-3-ol) moiety, and one new steroid (27), along with five known compounds (28¡V32) from Sinularia gibberosa. Three new cembrane-type diterpenoids (33¡V35), along with seven known cembranolides (36¡V42) were isolated from Sarcophyton sp. The structures of metabolites 1¡V42 including their stereochemistry 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 from other known compounds. In above metabolites, two compounds (8, 9) exhibited cytotoxicity against the growth of MCF-7, Hep 3B, Ca9-22 cancer cell lines. Furthermore, nine compounds (4, 8, 9, 11, 12, 13, 21, 31, 39) exhibited cytotoxicity against the growth of MDA-MB-231, Hep G2 and A-549 cancer cell lines.

In order to search for bioactive compounds, we have studied the chemical constituents from the organic extracts of the soft coral Sarcophyton crassocaule. This study had led to the isolation of twenty-six natural cembrane-type diterpenoids, compounds 1¡V26, including eighteen new compounds, sarcocrassocolide A¡VR (1¡V18), along with six know compounds, crassocolide A, B, D, E, L, sarcocrassolide, sinularolide E and 13-acetoxysarcocrassolide (19¡V26). The structures of compounds 1¡V26 were established by detailed spectroscopic data analysis (IR, MS, 1D, 2D NMR) and by comparison of the spectral data with those of the related known compounds. The structures of 8, 9 and 11 were further established by orgamic methods, and the absolute configuration of 1 was determined using a modified Mosher¡¦s method. The cytotoxicity of compounds 1¡V17 and 19¡V21 against the Daoy (human medulloblastoma), HEp2 (human laryngeal carcinoma), MCF-7 (human breast adenocarcinoma), WiDr (human colon adenocarcinoma), DLD-1 (human colon adenocarcinoma), CCRF-CEM (human T-cell acute lymphoblastic leukemia), and HL-60 (human promyelocytic leukemia) tumor cell lines were determined, and structure-activity relationship was presented by statistic method. Compounds 3 and 9 showed significant activity toward the above Daoy, HEp2, MCF7 and WiDr, and compounds 18, 19, 20, 22 and 24 were found to show significant activity toward the above DLD-1, CCRF-CEM and HL-60. Compounds 1¡V26 were shown to exert significant anti-inflammatory activity in LPS-stimulated RAW264.7 macrophage cells. Compounds 9, 17, 19, 22 and 24 also significantly inhibited the accumulation of pro-inflammatory COX-2 protein.

In order to search for bioactive compounds, we have studied the chemical constituents from the organic extracts of the soft coral Sarcophyton stellatum, collected from Dongsha Atoll. This study had led to the identification of sixteen natural diterpenoids, including six new compounds, stellatumonone (1), stellatumolides A¡VC (2¡V4) and stellatumonins A¡VB (5¡V6), along with ten known compounds. The structures of compounds 1¡V16 were established by detailed spectroscopic data (IR, MS, 1D, 2D NMR) and by comparison of the spectral data with those of the related known compounds. The cytotoxicity of compounds 1¡V16 against the HepG2 (human liver hepatocellular carninoma cells), MDA-MB231 (human breast cancer cells) and A549 (human respiratory epithelial cells) cancer cell lines were determined. Among them, compound 16 showed cytotoxic activity toward the A549 cancer cells.

In the course of studying on secondary metabolites from marine organisms, we have investigated the chemical constituents of the soft coral Sarcophyton ehrenbergi collected at San-Hsien-Tai, Taitong County, Taiwan. Chromatographic separation of the organic extracts has led to the isolation of nine new cembrane diterpenes 1¡V9, and a initial natural separation of known cembrane diterpene 10. The chemical structures of pure compounds were determined by spectral (NMR, MS, UV and IR) and physical data, as well as comparison with the spectroscopic data of related chemicals in literature. Moreover, the metabolites 1¡V10 were evaluated in vitro for their cytotoxicity against of A-549 (human lung epithelial carcinoma), HT-29 (human colon adenocarcinoma), and P-388 (mouse lymphocytic leukemia) cells, as well as anti-HCMV activity. Compounds 3, 6, 9, and 10 were shown to exhibit significant cytotoxicity activities against P-388 with ED50 values of 2.7, 3.6, 2.0, and 3.0 £gg/mL, respectively. Compound 1, 2, 3, and 7 exhibited inhibitory activity against anti-HCMV, with EC50 values of 60.0, 46.0, 5.0, and 45.0 £gg/mL.

The methylene chloride extracts of Formosan soft coral Sarcophyton crassocule Von. Marenzeller (collected at Green Island off Taiwan) were found to exhibit significant cytotoxicity against P-388, HT-29, and A-549 cancer lines. Chromatographic separation led to the isolation of three known cembrane diterpenoids, GN16-34 (1), GN16-35 (2), GN16-40 (3), and a new cembrane, GN16-62 (4). The methylene chloride and acetone extracts of Formosan soft coral Xenia puerto-galerae Roxas (collected at Green Island off Taiwan) were found to exhibit significant cytotoxicity against P-388 and A-549 cancer cell lines. Chromatographic separation resulted in the isolation of six new cadinane sesquiterpenes, GN44-28 (5), GN44-30 (6), GN44-44 (7), GN44-66 (8), GN44-173 (9), GN44-149 (10), and one known dicarbocyclic diterpenes with a bicyclic [4.3.1] ring system, GN44-199 (11). Compounds 3 and 4 exhibited significant cytotoxicity against P-388, HT-29, and A-549 cancer cell lines. Compounds 1 and 2 exhibited significant cytotoxicity against P-388 cancer cell lines. Compounds 5 and 7 exhibited significant cytotoxicity against A-549 cancer cell line. compound 10 exhibited significant cytotoxicity against P-388 and A-549 cancer cell lines.

Corals are the dominant species of the coral reefs. The diversity of species is classified by traditional morphological traits, especially relied on the calcious deposits-sclerites. The formation of sclerites may be affected by the environmental conditions; therefore, some controversies may exist. The ambiguities may be clarified with molecular approaches. Five soft coral species, Lobophytum pauciflorum, Sinularia leptoclado, Sinularia flexibilis, Sarcophyton sp., Nephthea erecta were chosen for this study. The total mitochondrial DNA sequences were determined by PCR and primer walking. The genome contains 18562 bp, 18732 bp, 18752 bp, 18806 bp, 18716 bp separately, which harbors 14 protein-coding genes (ATP6¡BATP8¡BCOI¡BCOII¡BCOIIII¡BCYTB¡BND1¡BND2¡BND3¡BND4¡BND4L¡BND5¡BND6¡BMSH), 2 rRNA and only 1 tRNA genes. The phylogenetic relationship of alcyonacea corals were analyzed and compared with published sequences. The possibility of using a short ¡§DNA bar-code sequence¡¨ of the mitochondria as an alternative for species identification may be feasible. We found the short DNA signature sequences for these five corals. They may speed up the identification of corals in the long run.