- freely available
Marine Drugs 2004, 2(4), 154-163;
Abstract: A new stereoisomer of an araguspongine/xestospongin alkaloid, named araguspongine M (1), has been isolated together with 12 known compounds, araguspongines B (2) and D (3), dopamine, three galactosyl diacylglycerols, 24-methyl cholesterol, 5,6-dihydrocholesterol, β-sitosterol, and three 5α,8α-epidioxy sterols (11–13), from the marine sponge Neopetrosia exigua (formerly Xestospongia exigua) collected in Palau. The structure of 1 was assigned on the basis of its spectral data analysis. This is the first report on the isolation of dopamine from a marine sponge. This compound may be produced by an endosymbiotic Synechococcus-like cyanobacterium. Compounds 1–3 and 11–13 showed cytotoxicity against HL-60 at IC50’s of 5.5, 5.5, 5.9, 22.4, 9.5, and 9.6 μM, respectively. The possible biosynthesis origin of the isolated metabolites is discussed.
Marine sponges (Porifera) are rich sources of bioactive compounds. It has also been well known that they harbor bacteria in their tissues . In some cases, the associated microorganisms constitute about a half of the biomass [2, 3]. Moreover, numbers of marine natural products obtained from sponges have been found to show structural similarities to the metabolites of marine and terrestrial microorganisms. These evidences presented the question whether the bioactive compounds are produced by the sponges or by the associated microorganisms . A few studies have revealed the localization of compounds previously obtained from sponges in the symbiotic microorganisms and the involvement of bacteria in the biosynthesis of the compounds .
We have started the chemical ecology study on the sponge Neopetrosia exigua, previously named as Xestospongia exigua, and describe here the constituents of the Palauan species. A new stereoisomer of an araguspongine/xestospongin alkaloid, named araguspongine M (1), and 12 known compounds (2–13) were obtained from the EtOH extract. It should be noted that this is the first report on the isolation of dopamine from a marine sponge.
Results and discussion
N. exigua is a common and widely distributed sponge in the shallow lagoon of Palau. A Synechococcus-like cyanobacterium and several types of bacteria were detected in the tissue by visible and fluorescent microscopic observations . The EtOH extract of the sponge was partitioned between water and 1-butanol. The butanol extract was subjected to repeated chromatographic separations to give the new compound, araguspongine M (1), araguspongines B (2) and D (3) [7, 8], dopamine (4), three galactosyl diacylglycerols (5–7) [9–13], 24-methyl cholesterol (8), 5, 6-dihydrocholesterol (9), β-sitosterol (10) [14–16], and three 5α,8α-epidioxy sterols (11–13) [17, 18]. The structures of known compounds were determined on the basis of their spectral data and comparison with the previously reported values.
Araguspongine M (1) was obtained as colorless tiny needles. The molecular formula C28H50N2O2 was deduced from the HRFABMS and NMR spectral data. The 1H and 13C NMR spectra of 1 resembled those of 2 and 3. The 13C NMR spectrum of 1 showed 14 signals ascribed to three methines and 11 methylenes (Table 1) as similar to those of 2 and 3, which suggested that 1 has a C2-symmetric structure. There are only two compounds (2 and 3) thus far isolated from sponges possessing the molecular formula of C28H50N2O2 and C2-symmetric structures. Therefore, 1 was revealed to be a new compound of aragustongine/xestospongin alkaloids [7, 8, 19–23].
Compound 3 showed the characteristic Bohlmann absorptions at 2754 and 2812 cm−1 in its IR spectrum. However, these bands were not detected in the IR spectra of 1 and 2, indicating that 1 have cis-fused 1-oxaquinolizidine rings as similar to the structure of 2. The 1H and 13C NMR signals of 1 were assigned by the analysis of 1H–1H COSY and HMQC spectral data and comparison with the signals of 2 and 3. The 1H–1H COSY spectrum of 1 revealed the carbon sequences of 2–3–4 and 6–7–8–9–10 together with the geminal couplings of H2-3, H2-4, H2-6, H2-7, and H2-8. The larger coupling constant observed for H-10 (δ 4.09, J = 10.0 Hz) indicated that H-9 is axially oriented. H-2 with a larger coupling constant (3.68, J = 10.8 Hz) is also at axial orientation confirmed by the NOE correlations observed between H-2 and H-9 in the NOESY and ROESY spectra of 1. These data suggested that 1-oxaquinolizidine rings in 1 have the ααα-II type structure reported by Hoye et al. . Consequently, the structure of araguspongine M was assigned as 1.
Araguspongine/xestospongin alkaloids have been principally detected in the genus Xestospongia (Neopetrosia), more precisely, N. exigua collected at different areas, such as Australia [20, 21], Red Sea , Okinawa Japan , India , Philippines , and Palau  (this study). While, araguspongine A (xestospongin D) was found in Niphates sp. from Singapore . A variety of bioactivities have been reported for araguspongines and xestospongins, such as inhibition of rat brain nitric oxide synthase activity , vasodilation activity [7, 21], cytotoxicity , antifungal activity , somatostatin and vasoactive intestinal peptide inhibition activity , antimalarial and antituberculosis activities , and inhibition of platelet aggregation .
Dopamine (4) was obtained in rather higher yield (0.0022% of wet weight). This is the first report of the isolation of dopamine from a marine sponge. Sponges are known as primitive animals without a nerve system, and thus dopamine may not be produced by the sponge as a neurotransmitter. L-Dopa-3-sulfate has been isolated from the brown alga Ascophyllum nodosum , and recent reports suggested that dopamine act as a grazer deterrent in the green alga Ulvaria obscura  and be involved in the allellopathic properties of U. obscura and Ulva fenestrate . TLC analysis of the extracts of 11 different marine sponges collected in Palau showed that Dopamine only contained in N. exigua. Therefore, dopamine isolated from N. exigua could be produced by a photosynthetic associate to show deterrent and antifouling effects, and correspondingly contribute to the predominance of N. exigua in that region. In fact, numerous Synechococcus-like cells were observed under a microscope at the outer tissue layer of the sponge . A series of sulfur-containing dopamine derivatives were also discovered in the ascidians of the genera Lissoclinum, Aplydium, Eudistoma, and Polycitor [33–36]. These compounds or their precursors may be produced by the symbiotic microorganism.
Galactosyl diacylglycerols are widely distributed in plants as structural components of the thylakoid membrane. These compounds were also isolated from marine sponges, Phyllospongia foliascens  and Pseudoceratina sp. , and marine microorganisms, such as cyanobacteria [9–10], dinoflagellates [11, 12], and a lichenized fungus . HPLC analysis of the glycolipids fractions revealed that there were at least seven minor monogalactosyl and digalactosyl diacylglycerols, and several trigalactosyl diacylglycerols in this sponge. Compound 5 was the mostly abundant component among them. These lipids would be produced by the photosynthetic symbiont (most likely a Synechococcus-like cyanobacterium) .
Sterols possessing the 5α, 8α-epidioxy moiety have been found in eight marine sponges of different genera, Axinella cannabina, Tethya aurantia, Raphidostila incise, Thalyisias juniperina, Haliclona rubens, Hyrtios sp., Axinissa sp., Dysidea fragilis, and Luffariella cf. variabilis . This report revealed a new source of these sterols. Therefore, the question has been raised whether these sterols are produced by the sponges or by the associated microorganisms.
Separation of microorganisms and sponge cells from N. exigua, cultivation of these cells, and analysis of genes for the biosynthesis of the secondary metabolites are now under investigation.
Cytotoxicity against the human leukemia cell line HL-60 and antimicrobial activity of compounds 1–13 were examined. The IC50 values of compounds 1, 2, 3, 11, 12, and 13 against HL-60 were 5.5, 5.5, 5.9, 22.4, 9.5, and 9.6 μM, respectively. Compounds 1–13 did not inhibit the growth of Escherichia coli, Staphylococcus aureus, Saccharomyces cerevisiae, Mucor hiemalis, and marine bacterium Ruegeria atlantica even at 100 μg/disc.
A chemical ecology study of the marine sponge Neopetrosia exigua resulted in the isolation of a new stereoisomer of an araguspongine/xestospongin alkaloid, named araguspongine M (1), together with 12 known compounds. This is also the first report of the isolation of dopamine from a marine sponge. Dopamine may be produced by an endosymbiotic Synechococcus-like cyanobacterium to show deterrent and antifouling effects. Compounds 1–3 and 11–13 showed cytotoxicity against HL-60 cell line, and all isolates were inactive against the five test microorganisms.
Optical rotations were obtained on a JASCO DIP-130 polarimeter. UV and IR spectra were recorded on a Hitachi U-3000 and on a JASCO A-102, respectively. NMR spectra were measured on either a JEOL JNM ECP-600, A-500, or AL-400 NMR spectrometer. Mass spectra were obtained by either a JEOL HX-110 mass spectrometer (FAB mode, m-nitrobenzylalcohol as matrix) or a Finnigan TSQ 700 triple quadrupole mass spectrometer (ESI mode).
N. exigua was collected by skin diving at Iwayama Bay in Palau during the training vessel Shin’yo Maru was anchored at the Marakal Port in March, 2003. The sponge was kept in a freezer at −50 °C on the ship and transported to Japan. The voucher specimen is deposited at the Department of Ocean Sciences, Tokyo University of Marine Science and Technology, as TUF number 03-03-04=2–23. The sponge was identified by Professor P. Bergquist (University of Auckland, New Zealand) .
Extraction and Isolation
The sponge (900g, wet weight) was thawed, cut into small pieces, and extracted with EtOH (3L × 3). The extract was evaporated and partitioned between water and 1-BuOH (saturated with water). The BuOH extract (9.0 g) was separated on an ODS column (eluted with MeOH) to afford four fractions (Fr. B-1–Fr. B-4). Fr. B-1 (1.2 g) was separated by an HW-40 column (MeOH) into two fractions (B-1-1 and B-1-2). Fr. B-1-1 was subjected to ODS column (100% H2O to100% MeOH) and HPLC (ODS, 0.1% TFA/H2O) separations to give dopamine (4, 20 mg). Araguspongines M (1, 6.5 mg), B (2, 6.0 mg), and D (3, 5.6 mg) were isolated from Fr. B-1-2 by SiO2 (CHCl3–MeOH–NH4OH, 30:1:0.05, 20:1:0.2, 10:1:0.5) followed by LH-20 (MeOH) column chromatographies. Fr. B-2 (0.9g) was separated by a SiO2 column (CHCl3–MeOH–H2O, 90:10:0.5, 80:20:1, 70:30:5) and then HPLC on ODS (100% MeOH) to give compounds 5 (50 mg), 6 (3.7 mg), and 7 (12.0 mg). Fr. B-4 (2.5g) was subjected to a SiO2 column (CHCl3–MeOH, 100:0, 100:1, 50:1, 25:1), an LH-20 column (MeOH), and HPLC (ODS, 100% MeOH) separations to afford 8 (2.2 mg), 9 (30 mg), 10 (170 mg), 11 (9.3 mg), 12 (3.2 mg), and 13 (1.4 mg).
Araguspongine M (1): [α]D −4.6° (c 0.5, CHCl3); IR (KBr) λmax 3437, 2934, 2857, 2733, 2072, 1638, 1465, 1420, 1173, 1131, 1092, 1040 cm−1; 1H and 13C NMR data (CDCl3) see Table 1; HRFABMS m/z 447.3932 [M + H]+ (calcd for C28H51N2O2, 447.3951).
Araguspongine B (2): [α]D −3.6° (c 0.5, CHCl3); IR (KBr) λmax 3421, 2927, 2850, 1454, 1350, 1141, 1095, 1033, 964, 910 cm−1; ESIMS m/z 447 ([M+H]+).
Araguspongine D (3): [α]D −10.3° (c 0.5, CHCl3); IR (KBr) λmax 3421, 2931, 2850, 2812, 2754, 2360, 2341, 1465, 1346, 1276, 1157, 1134, 1095, 1037, 948cm−1; ESIMS m/z 447 ([M+H]+).
TLC analysis of dopamine in marine sponges
Freeze dried small pieces of 11 species of marine sponges including N. exigua were each extracted with 50% EtOH. The extract was evaporated, and the residue was redissolved with 50% EtOH to adjust the concentration at 1 g dry sponge weight/mL. Five microliter of each extract was loaded on a SiO2 TLC plate together with 2 μL of dopamine HCl (1 mg/mL) as control. TLC plates were developed with 1-BuOH–AcOH–H2O (4:1:2, Rf = 0.45) and detected with the Ninhydrin reagent.
The human leukemia cell line HL-60 was incubated in RPMI 1640 using 24-well assay plates. Samples were dissolved in MeOH, and 10 μL of each sample was poured in a well and the solvent evaporated in a clean bench. The suspension (1 mL, 4 × 104 cells/mL) of HL-60 was added to each well and incubated at 37 °C for 72 hours in a CO2 incubator. The shape of the cells was observed after 72 hours under an inverted microscope. The number of vital cells in the sample wells after 72 hours was compared with those in the control wells using XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] (cell proliferation kit II®).
The growth inhibitory activity of samples was evaluated by a paper disc method against Escherichia coli IAM 12119T, Staphylococcus aureus IAM 12544T, Saccharomyces cerevisiae IAM 1438T, Mucor hiemalis IAM 6088, and the marine bacterium Ruegeria atlantica TUF-D as described in the previous report .
|Atom No.||13C||1H (J in Hz)|
|2||76.4||3.68 br t (10.8)|
|3||29.5||1.74 m, 2.26 m|
|4||52.7||3.01 m, 3.54 br d (10.4)|
|6||53.3||2.73 m, 3.35 br d (11.2)|
|7||22.2||1.72 m, 2.33 m|
|8||24.8||1.30 m, 1.43 m|
|10||94.9||4.09 d (10.0)|
|11||30.5||1.36 m, 1.64 m|
|12||28.7||1.14 m, 1.32 m|
|13||31.1||1.16 m, 1.32 m|
|14||27.3||1.78 m, 1.84 m|
|15||24.3||1.14 m, 1.27 m|
|16||34.5||1.48 m, 1.70 m|
aSignals were assigned by 1H-1H COSY and HMQC experiments and comparison with the NMR data for 2 and 3.
We thank Drs. M. Endo and Y. Shizuri of the Marine Biotechnology Institute, Japan, for the sample collection in Palau and for the information on the identification of the sponge. We are grateful to the crewmembers of the training vessel Shin’yo Maru operated by Tokyo University of Marine Science and Technology for their generous help for sample collection at Palau.
- Sample Availability: Samples are available from the authors.
References and Notes
- Wilkinson, C. R. Significance of microbial symbionts in sponge evolution and ecology. Symbiosis 1987, 4, 135–146. [Google Scholar]
- Vacelet, J.; Donadey, C. Electron microscope study of the association between some sponges and bacteria. J. Exp. Mar. Ecol 1977, 30, 301–314. [Google Scholar]
- Willenz, P.; Hartman, W. D. Micromorphology and ultrastructure of Caribbean sclerosponges. I. Ceratoporella nicholsoni and Stromatospongia norae (Ceratoporellidae: Porifera). Mar. Biol 1989, 103, 387–402. [Google Scholar]
- Haygood, M. G.; Schmidt, E. W.; Davidson, S. K.; Faulkner, D. J. Microbial symbionts of marine invertebrates: Opportunities for microbial biotechnology. J. Mol. Microbiol. Biotechnol 1999, 1, 33–43. [Google Scholar]
- Faulkner, D. J.; Harper, M. K.; Haygood, M. G.; Salomon, C. E.; Schmidt, E. W. Symbiotic bacteria in sponges: Sources of bioactive substances. In Drugs from the Sea; Fusetani, N., Ed.; Kaeger: Basel, 2002; pp. 107–119. [Google Scholar]
- Thin slices and homogenized samples were prepared from N. exigua and observed under a microscope in a laboratory at the Palau International Coral Reef Center in March, 2004.
- Kobayashi, M.; Kawazoe, K.; Kitagawa, I. Araguspongines B, C, D, E, F, G, H, and J, new vasodilative bis-1-oxaquinolizidine alkaloids from an Okinawan marine sponge, Xestopongia sp. Chem. Pharm. Bull 1989, 37, 1676–1678. [Google Scholar]
- Kobayashi, M.; Miyamota, Y.; Aoki, S.; Murakami, N.; Kitagawa, I.; Ishida, T. Isomerization of dimeric 2, 9-disubstituted 1-oxaquinolizidine alkaloids and structural revision of araguspongines B and E, isolated from a marine sponge of Xestospongia sp. Heterocycles 1998, 47, 195–203. [Google Scholar]
- Morimoto, T.; Murakami, N.; Nagatsu, A.; Sakakibara, J. Studies on glycolipids VII. Isolation of two new sulfoquinovosyl diacylglycerols from green alga Chlorella vulgaris. Chem. Pharm. Bull 1993, 41, 1545–1548. [Google Scholar]
- Shirahashi, H.; Murakami, N.; Watanabe, M.; Nagatsu, A.; Sakakibara, J.; Tokuda, H.; Nishino, H.; Iwashima, A. Isolation and identification of anti-tumor-promoting principles from the fresh-water cyanobacterium Phormidium tenue. Chem. Pharm. Bull 1993, 41, 1664–1666. [Google Scholar]
- Oshima, Y.; Yamada, S. H.; Matsunaga, K.; Moriya, T.; Ohizumi, Y. A monogalactosyl diacylglycerol from a cultured marine dinoflagellate, Scrippsiella trochoidea. J. Nat. Prod 1994, 57, 534–536. [Google Scholar]
- Kobayashi, M.; Hayashi, K.; Kawazoe, K.; Kitagawa, I. Marine natural products. XXIX. Heterosigma-glycolipids I, II, III, and IV, four diacylglyceroglycolipids possessing ω3-polyunsaturated fatty acid residues, from the raphidophycean dinoflagellate Heterosigma akashiwo. Chem. Pharm. Bull 1992, 40, 1404–1410. [Google Scholar]
- Sassaki, G. L.; Gorin, P. A. J.; Iacomini, M. Characterization of lyso-galactolipids, C-2 and C-3 O-acyltrigalactosylglycerol isomers, obtained from the lichenized fungus Dictyonema glabratum. FEMS Microbiol. Lett 2001, 194, 155–158. [Google Scholar]
- Huang, P.; Karagianis, G.; Waterman, P.G. Chemical constituents from Typhonium flagelliforme. Zhong Yao Cai 2004, 27, 173–175. [Google Scholar]
- Xu, X.; Guan, Z.; Zeng, L.; Su, J. Study on the steroid constituents of soft coral Lobophytum microspiculatum. Se Pu 1999, 17, 225–228. [Google Scholar]
- Blumenberg, M.; Thiel, V.; Pape, T.; Michaelis, W. The steroids of hexactinellid sponges. Naturwissenschaften 2002, 89, 415–419. [Google Scholar]
- Gauvin, A.; Smadja, J.; Aknin, M.; Faure, R.; Gaydou, E. M. Isolation of bioactive 5α, 8α-epidioxy sterols from the marine sponge Luffariella cf. variabilis. Can. J. Chem 2000, 78, 986–992. [Google Scholar]
- Gunatilaka, A. A. L.; Gopichand, Y.; Francis, J. S.; Djerassi, C. Isolation and structure elucidation of nine new 5α,8α-epidoxy sterols from four marine organisms. J. Org. Chem 1981, 46, 3860–3866. [Google Scholar]
- Orabi, K. Y.; El Sayed, K. A.; Hamann, M. T.; Dunbar, D. C.; Al-Said, M. S.; Higa, T.; Kelly, M. Araguspongines K and L, new bioactive bis-oxaquinozilidine N-oxide alkaloids from red sea specimens of Xestospongia exigua. J. Nat. Prod 2002, 65, 1782–1785. [Google Scholar]
- Moon, S. S.; Macmillan, J. B.; Olmstead, M. M.; Ta, T. A.; Pessah, I. N.; Molinski, T. F. (+)-7S-hydroxyxestosponin A from the marine sponge Xestospongia sp. and absolute configuration of (+)-xestospongin D. J. Nat. Prod 2002, 65, 249–254. [Google Scholar]
- Nakagawa, M.; Endo, M.; Tanaka, N.; Lee, G.-P. Structures of xestospongin A, B, C and D, novel vasodilative compounds from marine sponge, Xestospongia exigua. Tetrahedron Lett 1984, 25, 3227–3230. [Google Scholar]
- Reddy, M. V. R.; Faulker, D. J. 3β, 3′β,-dimethylxestospongin C, a new bis-1–oxa quinolizidine alkaloid from the Palauan sponge Xestospongia sp. Nat. Prod. Lett 1997, 11, 53–59. [Google Scholar]
- Venkateswarlu, Y.; Reddy, M. V. R.; Rao, J. V. Bis-1-oxaquinolizidine from the sponge Haliclona exigua. J. Nat. Prod 1994, 57, 1283–1285. [Google Scholar]
- Bohlmannn, F. Zur konfigurationsbestimmung von chinolizin-derivaten. Angew. Chem 1957, 69, 641–644. [Google Scholar]
- Hoye, T. R.; North, J. T.; Yao, L. J. Conformational considerations in 1-oxaquinolizidines related to the Xestospongin/araguspongine family: Reassignment of stereostructures for araguspongines B and E. J. Org. Chem 1994, 59, 6904–6910, Structure correction of araguspongine B, J. Org. Chem. 1995, 60, 4958. [Google Scholar]
- Rao, J.; Desaiah, D.; Vig, P.; Venkateswarlu, Y. Marine biomolecules inhibit rat brain nitric oxide synthase activity. Toxicology 1998, 129, 103–112. [Google Scholar]
- Pettit, G.; Orr, B.; Herald, D.; Doubek, D.; Tackett, L.; Schmidt, J.; Boyd, M.; Pettit, R.; Hooper, J. Isolation and X-ray crystal structure of racemic xestospongin D from the Singapore marine sponge Niphates sp. Bioorg. Med. Chem. Lett 1996, 6, 1313–1318. [Google Scholar]
- Vassas, A.; Bourdy, G.; Paillard, J.; Lavayre, J.; Pais, M.; Quirion, J.; Debitus, C. Naturally occurring somatosatatin and vasoactive intestinal peptide inhibitors: Isolation of alkaloids from two marine sponges. Planta Med 1996, 62, 28–30. [Google Scholar]
- Pimentel, S. M. V.; Bojo, Z. P.; Roberto, A. V. D.; Lazaro, J. E. H.; Mangalindan, G. C.; Florentino, L. M.; Lim-Navarro, P.; Tasdemir, D.; Ireland, C. M.; Concepcion, G. P. Platelet aggregation inhibitors from Philippine marine invertebrate samples screened in a new microplate assay. Mar. Biotechnol 2003, 5, 395–400. [Google Scholar]
- Laycock, M. V.; Rogan, M. A. L-3, 4-Dihydroxyphenylalanine-3-sulfate from the brown alga, Ascophyllum nodosum. J. Nat. Prod 1984, 47, 1033–1036. [Google Scholar]
- Nelson, A. V. Chemical Defenses of Ulvaria obscura: Effect on Food Preference of Strongylocentrotus droebachiensis. M.S. Thesis, Western Washington University, Bellingham, WA, 2003; p. 49. [Google Scholar]
- Nelson, T. A.; Lee, D. J.; Smith, B. C. Are green tides harmful algal blooms? Toxic properties of water-soluble extracts from two bloom-forming macroalgae, Ulva fenestrata and Ulvaria obscura (Ulvophyceae). J. Phycol 2003, 39, 874–879. [Google Scholar]
- Davidson, B. S.; Molinski, T. F.; Barrows, L.; Ireland, C. M. Varacin: a novel benzopentathiepin from Lissoclinum vareau that is cytotoxic toward a human colon tumor. J. Am. Chem. Soc 1991, 113, 4709–4710. [Google Scholar]
- Litaudon, M.; Trigalo, F.; Martin, M. T.; Frappier, F.; Guyot, M. Lissoclinotoxins: Antibiotic polysulfur derivatives from the tunicate Lissoclinum perforatum. Revised structure of lissoclinotoxin A. Tetrahedron 1994, 50, 5323–5334. [Google Scholar]
- Compagnone, R. S.; Faulkner, D. J.; Carté, B. K.; Chan, G.; Freyer, A.; Hemling, M. E.; Hofmann, G. A.; Mattern, M. R. Pentathiepins and trithianes from two Lissoclinum species and a Eudistoma sp.: inhibitors of protein kinase C. Tetrahedron 1994, 50, 12785–12792. [Google Scholar]
- Searle, P. A.; Molinski, T. F. Five new alkaloids from the tropical ascidian, Lissoclinum sp. lissoclinotoxin A is chiral. J. Org. Chem 1994, 59, 6600–6605. [Google Scholar]
- Kikuchi, H.; Tsukitani, Y.; Manda, T.; Fujii, T.; Nakanishi, H.; Kobayashi, M.; Kitagawa, I. Marine natural products X. Pharmacologically active glycolipids from the Okinawan marine sponge Phyllospongia foliascens. Chem. Pharm. Bull 1982, 30, 3544–3547. [Google Scholar]
- Meguro, S.; Namikoshi, M.; Kobayashi, H. A new screening method for antimitotic substances and isolation of glycolipids as stimulators of tubulin polymerization form Okinawan sponge Pseudoceratian sp. J. Antibiot 2002, 55, 256–262. [Google Scholar]
- The voucher specimen of the same sponge is also kept in the Marine Biotechnology Institute, Japan as 02PA-006, which was identified by Professor P. Bergquist as Neopetrosia exigua Kirkpatrick (Order Petrosida, Family Petrosiidae).
- Namikoshi, M.; Negishi, R.; Nagai, H.; Dmitrenok, A.; Kobayashi, H. Three new chlorine containing antibiotics from a marine-derived fungus Aspergillus ostianus collected in Pohnpei. J. Antibiot 2003, 56, 755–761. [Google Scholar]
© 2004 by MDPI Reproduction is permitted for noncommercial purposes.