針對採集自南臺灣綠島海域的柳羽珊瑚 Pinnigorgia sp. 的化學組成分析中，分離出 5 個新的 4,5;5,6;9,11-triseco-4(5→6),5(6→7)-diabeo-ergostane 型式的固醇類化合物，pinnigorgiols A−E (1−5) 及11 個新 9,11-secosteroid 型式的固醇類化合物 pinnisterols A−K (6−16) ， 5 個新固醇類化合物， (22R)-acetoxy-24-methylcholest-5-ene-3β,25-diol (17) 、(22E,24R)-ergosta-5,22-diene- 3β,11α-diol(18) 、(24S)-ergosta-5-ene-3β,11α-diol (19) 、5α,6α-epoxy-23-dimethyl-gorgost-8-ene-3β,7α-diol (20)、5α,6α-epoxy-23-demethylgorgost-8(14)- ene-3β,7α-diol (21)，4 個已知固醇類化合物 cerevisterol (22)、23-demethylgorgost- 7-ene-3,5,6-triol (23)、ergosterol peroxide (24) 、(22R,23R,24R)-5,8-epidioxy-22,23-methylene-24-methylcholest-6-en-3-ol (25) 與已知化合物 apo-9′-fucoxanthinone (26) 和pubinernoid A (27)。化合物的結構均藉由光譜分析及文獻資料比對方法加以解析建立，特別是一維和二維核磁共振實驗。化合物 1−5 更藉由圓二色光譜 (CD spectrum) 比較其立體構型。

Studies on the chemical constituents of a gorgonian coral identified as Pinnigorgia sp. have resulted in the isolation of 5 new 4,5;5,6;9,11-triseco-4(5→6),5(6→7)-diabeo-ergostane, pinnigorgiols A−E (1−5); 11 new 9,11-secosteroids, pinnisterols A−K (6−16); five new sterols, (22R)-24-methyl-5-ene-3β-ol-22-acetate (17), (22E,24R)- ergosta-5,22-diene-3β,11α-diol (18), (24S)-ergosta-5-ene-3β,11α-diol (19), 5α,6α- epoxy-23-demethylgorgost-8(14)-ene-3β,7α-diol (20), 5α,6α-epoxy-23-demethyl- gorgost-8-ene-3β,7α-diol (21); and six known compounds, cerevisterol (22), 23-demethylgorgost-7-ene-3,5,6-triol (23), ergosterol peroxide (24), (22R,23R,24R)-5,8-epidioxy-22,23-methylene-24-methylcholest-6-en-3-ol (25), apo-9′-fucoxanthinone (26), and pubinernoid A (27). The structures of each metabolite were determined by spectroscopic methods, particular by the 1D and 2D NMR experiments. The configurations of metabolite 1−5 were further confirmed by comparative analysis of circular dichroism spectroscopic data.
Each metabolite was screened for anti-hepatofibric and anti-inflammatory activities. Metabolites 1, 2, 4−6, 9, 10, 12, 13, and 23 were found to exhibit cytotoxicity in HSC-T6. Activation of hepatic stellate cells (HSCs) plays a major role in liver fibrosis. During liver fibrosis, extracellular matrix proteins and pro-inflammatory growth factors are overexpressed by activated HSCs. Clearance of activated HSCs by inducing apoptosis is an efficient way to treat liver fibrosis. In the anti-inflammation assay, compounds 1−6, 8, 9, 12, and 15 inhibited the production of superoxide anion and decreased the release of elastase in the human neutrophils that stimulated by formyl-L-methionyl-L-leucyl- L-phenylalanine/cytochalasin B (fMLP/CB). The metabolite 13 only caused the inhibit superoxide anion generation, while the compounds 11 and 26 only resulted in inhibition of elastase release in human neutrophils. Although numerous studies have attempted to characterize marine bioactive compounds with the aim of curing human diseases, the focus on liver diseases is very limited. Further biomedical investigation of the 9,11-secosteroids may warrant the application to be promising bioactive agents for liver disease.

Verification letter from the Oral Examination Committee i
Acknowledgements ii
Abstract in Chinese iii
Abstract in English iv
The structures of compounds 1－27 vi
Contents viii
List of Figures xi
List of Tables xxii
I. Introduction 1
1.1. Development of natural product research 1
1.2. Marine organisms as sources of drug discovery 2
II. Literature review 7
2.1 Literature review : secosteroids of marine origin 7
III. Materials and Methods 44
3.1. Research design 44
3.2. The collected conditions, scientific classification of Pinnigorgia sp. 44
3.3. Extraction and isolation of Pinnigorgia sp. 45
3.4. Biological activities 52
3.4.1 Anti-hepatofibric assay 52
3.4.2 Human neutrophil superoxide anion generation and elastase release 52
3.5. General experimental procedures and meterials 53
3.5.1 General experimental procedures 53
3.5.2 General experimental solvents 55
IV. Compound identification 57
4.1 Characterization of pinnigorgiol A (1) 57
4.2 Characterization of pinnigorgiol B (2) 73
4.3 Characterization of pinnigorgiol C (3) 83
4.4 Characterization of pinnigorgiol D (4) 93
4.5 Characterization of pinnigorgiol E (5) 103
4.6 Characterization of pinnisterol A (6) 113
4.7 Characterization of pinnisterol B (7) 123
4.8 Characterization of pinnisterol C (8) 133
4.9 Characterization of pinnisterol D (9) 141
4.10 Characterization of pinnisterol E (10) 151
4.11 Characterization of pinnisterol F (11) 161
4.12 Characterization of pinnisterol G (12) 171
4.13 Characterization of pinnisterol H (13) 179
4.14 Characterization of pinnisterol I (14) 187
4.15 Characterization of pinnisterol J (15) 197
4.16 Characterization of pinnisterol K (16) 207
4.17 Characterization of (22R)-acetoxy-24-methylcholest-5-ene-3β,25-diol 217
4.18 Characterization of (22E,24R)-ergosta-5,22-diene-3β,11α-diol (18) 227
4.19 Characterization of (24S)-ergosta-5-ene-3β,11α-diol (19) 237
4.20 Characterization of 5α,6α-epoxy-23-demethylgorgost-8(14)-ene- 3β,7α- diol (20) 247
4.21 Characterization of 5α,6α-epoxy-23-demethylgorgost-8-ene-3β,7α- diol (21) 257
4.22 Characterization of cerevisterol (22) 267
4.23 Characterization of 23-demethylgorgost-7-ene-3,5,6-triol (23) 273
4.24 Characterization of ergosterol peroxide (24) 279
4.25 Characterization of (22R,23R,24R)-5α,8α-epidioxy-22,23-methylene- 24-methylcholest-6-en-3β-ol (25) 285
4.26 Characterization of apo-9′-fucoxanthinone (26) 291
4.27 Characterization of pubinernoid A (27) 295
V. Analyses of bioactivities 299
5.1. Result of anti-hepatofibric assay for compounds 1−27 299
5.2. Result of anti-inflammatory assay for compounds 1−27 301
VI. Conclusion 304
VII. Physical data of compounds 1−27 319
VIII. References 326

1. Barton, S.D.; Nakanishi, K. Comprehensive Natural Products Chemistry. Elsevier, New York. 1999.
2. Ruggieri, G.D. Drugs from the sea. Science 1976, 194, 491–497.
3. Mayer, A.M.S.; Glaser, K.B.; Cuevas, C.; Jacobs, R.S.; Kem, W.; Little, R.D.; McIntosh, J.M.; Newman, D.J.; Potts, B.C.; Shuster, D.E. The odyssey of marine pharmaceuticals: a current pipeline perspective. Trends. Pharmacol. Sci. 2010, 31, 255–265.
4. Hu, Y.; Chen, J.; Hu, G., Yu, J.; Zhu, X.; Lin, Y.; Chen, S.; Yuan, J. Statistical research on the bioactivity of new marine natural products discovered during the 28 years from 1985 to 2012. Mar. Drugs 2015, 13, 202-221.
5. Ebada, S.S.; Edrada, R.A.; Lin, W.; Proksch, P. Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates. Nat. Protoc. 2008, 3, 1820–1831.
6. Bao, B.; Zhang, P.; Lee, Y.; Hong, J.; Lee, C.O.; Jung, J.H., Monoindole alkaloids from a marine sponge Spongosorites sp. Mar. Drugs 2007, 5, 31–39.
7. Sohn, J.H.; Lee, Y.R.; Lee, D.S.; Kim, Y.C.; Oh, H. PTP1B inhibitory secondary metabolites from marine-derived fungal strains Penicillium spp. and Eurotium sp. J. Microbiol. Biotechnol. 2013, 23, 1206–1211.
8. Lillsunde, K.E.; Festa, C.; Adel, H.; de Marino, S.; Lombardi, V.; Tilvi, S.; Nawrot, D.A.; Zampella, A.; D’Souza, L.; D’Auria, M.V.; Tammela, P. Bioactive cembrane derivatives from the Indian Ocean soft coral, Sinularia kavarattiensis. Mar. Drugs 2014, 12, 4045–4068.
9. Gerwick, W.H.; Moore, B.S. Lessons from the past and charting the future of marine natural products drug discovery and chemical biology. Chem. Biol. 2012, 19, 85–98.
10. Rocha, J.; Peixe, L. Gomes, N.C.M.; Calado, R. Cnidarians as a source of new marine bioactive compounds—an overview of the last decade and future steps for bioprospecting. Mar. Drugs 2011, 9, 1860–1886.
11. D'Auria, M.V.; Minale, L.; Riccio, R. Polyoxygenated steroids of marine origin. Chemical Reviews 1993, 93, 1839–1895.
12. Sica, D.; Musumeci, D. Secosteroids of marine origin. Steroids 2004, 69, 743–756.
13. Madaio, A.; Piccialli, V.; Sica, D. Hipposterol, a unique trihydroxylated 5,6-secosterol from the marine sponge Hippospongia communis. Tetrahedron Lett. 1988, 29, 5999–6000.
14. Madaio, A.; Notaro, G.; Piccialli, V.; Sica, D. Minor 5,6-Secosterols from the marine sponge Hippospongia communis. Isolation and synthesis of (7Z,22E,24R)-24-methyl-5,6-secocholesta-7,22-diene-3β,5β,6-triol. J. Nat. Prod. 1990, 53, 565–572.
15. Valeria D'Auria, M.; Paloma, L.G.; Minale, L.; Riccio, R.; Debitus, C. Jereisterol A and B: Two 3β-methoxy-secosteroids from the pacific sponge Jereicopsis graphidiophora. Tetrahedron Lett. 1991, 32, 2149–2152.
16. Costantino, V.; Fattorusso, E.; Mangoni, A.; Aknin, M.; Gaydou, E.M. Novel 3-β-methoxysteroids from the senegalse sponge Microscleroderma spirophora. Steroids 1994, 59, 181–184.
17. Fouad, M.; Al-T.rabeen, K.; Badran, M.; Wray, V.; Edrada, R.; Proksch P.; Ebel, R. New steroidal saponins from the sponge Erylus lendenfeldi. ARKIVOC 2004, 8, 17–27.
18. Umeyama, A.; Shoji, N.; Enoki, M.; Arihara, S. Swinhosterols A−C, 4-Methylene Secosteroids from the Marine Sponge Theonella swinhoei. J. Nat. Prod. 1997, 60, 296–298.
19. Fusetani, N.; Nagata, H.; Hirota, H.; Tsuyuki, T. Astrogorgiadiol and astrogorgin, inhibitors of cell division in fertilized starfish eggs, from a gorgonian astrogorgia sp. Tetrahedron Lett. 1989, 30, 7079–7082.
20. Seo, Y.; Shin, J.; Song, J.-I. New Secosteroids from an undescribed gorgonian of the genus Muricella. J. Nat. Prod. 1995, 58, 1291–1295.
21. Seo, Y.; Cho, K.W.; Chung, H.; Lee, H.-S.; Shin, J. New secosteroids from a gorgonian of the genus Muricella. J. Nat. Prod. 1998, 61, 1441–1443.
22. Sun, X.-P.; Cao, F.; Shao, C.-L.; Chen, M.; Liu, H.-J.; Zheng, C.-J.; Wang, C.-Y. Subergorgiaols A–L, 9,10-secosteroids from the south China sea gorgonian Subergorgia rubra. Steroids 2015, 94, 7–14.
23. Lai, D.; Yu, S.; van Ofwegen, L.; Totzke, F.; Proksch, P.; Lin, W. 9,10-Secosteroids, protein kinase inhibitors from the Chinese gorgonian Astrogorgia sp. Bioorg. Med. Chem. 2011, 19, 6873–6880.
24. Li, T.-T.; Tang, X.-L.; Chen, C.-L.; Zhang, X.-W.; Wu, R.-C.; Zhu, H.-Y.; Li, P.-L.; Li, G.-Q. New eunicellin diterpenes and 9,10-Secosteroids from the gorgonian Muricella sibogae. Helv. Chim. Acta 2013, 96, 1188–1196.
25. Ochi, M.; Yamada, K.; Kotsuki, H.; Shibata, K. Calicoferols A and B, Two novel secosterols possessing brine-shrimp lethality from the gorgonian Calicogorgia sp. Chem. Lett. 1991, 20, 427–430.
26. Migliuolo, A.; Piccialli, V.; Sica, D. Structure elucidation and synthesis of 3β,6α-dihydroxy-9-oxo-9,11-seco-5α-cholest-7-en-11-al, a novel 9,11-secosterol from the sponge Spongia officinalis. Tetrahedron 1991, 47, 7937–7950.
27. Migliuolo, A.; Piccialli, V.; Sica, D. Two new 9,11-secosterols from the marine sponge Spongia officinalis. Synthesis of 9,11-seco-3β,6α,11-trihydroxy-5α- cholest-7-en-9-one. Steroids 1992, 57, 344–347.
28. Aknin, M.; Costantino, V.; Mangoni, A.; Fattorusso, E.; Gaydou, E. M. New 9,11-secosterols from gorgonia Subergorgia suberosa of the Indian Ocean. Steroids 1998, 63, 575–578.
29. Adinolfi, R.; Migliuolo, A.; Piccialli, V.; Sica, D. Isolation and synthesis of a new 9,11-Secosterol from the Sponge Spongia officinalis. J. Nat. Prod. 1994, 57, 1220–1226.
30. Pika, J.; Andersen, R. J. Blancasterol, a cytotoxic 9,11-secosteroid isolated from the northeastern pacific marine sponge Pleraplysilla sp. Tetrahedron 1993, 49, 8757–8760.
31. Lopp, A.; Pihlak, A.; Paves, H.; Samuel, K.; Koljak, R.; Samel, N. The effect of 9,11-secosterol, a newly discovered compound from the soft coral Gersemia fruticosa, on the growth and cell cycle progression of various tumor cells in culture. Steroids 1994, 59, 274–281.
32. Koljak, R.; Lopp, A.; Pehk, T.; Varvas, K.; Müürisepp, A.-M.; Järving, I.; Samel, N. New cytotoxic sterols from the soft coral Gersemia fruticosa. Tetrahedron 1998, 54, 179–186.
33. Koljak, R.; Pehk, T.; Järving, I.; Liiv, M.; Lopp, A.; Varvas, K.; Vahemets, A.; Lille, Ü.; Samel, N. New antiproliferative 9,11-secosterol from soft coral Gersemia fruticosa. Tetrahedron Lett. 1993, 34, 1985–1986.
34. Ioannou, E.; Abdel-Razik, A. F.; Alexi, X.; Vagias, C.; Alexis, M. N.; Roussis, V. 9,11-Secosterols with antiproliferative activity from the gorgonian Eunicella cavolini. Bioorg. Med. Chem. 2009, 17, 4537–4541.
35. Aiello, A.; Esposito, G.; Fattorusso, E.; Iuvone, T.; Luciano, P.; Menna, M. Aplidiasterols A and B, two new cytotoxic 9,11-secosterols from the mediterranean ascidian Aplidium conicum. Steroids 2003, 68, 719–723.
36. Aiello, A.; Fattorusso, E.; Menna, M.; Carnuccio, R.; Iuvone, T. New cytotoxic steroids from the marine sponge Dysidea fragilis coming from the lagoon of Venice. Steroids 1995, 60, 666–673.
37. Pika, J.; Tischler, M.; Andersen, R.J. Glaciasterols A and B, 9,11-secosteroids from the marine sponge Aplysilla glacialis. Can. J. Chem. 1992, 70, 1506–1510.
38. Reddy, M.V. R.; Harper, M.K.; Faulkner, D.J. Luffasterols A−C, 9,11- secosterols from the Palauan Sponge Luffariella sp. J. Nat. Prod. 1997, 60, 41–43.
39. Lu, Q.; Faulkner, D.J. Two new sesterterpenoids and a new 9,11-secosterol from Spongia matamata. J. Nat. Prod. 1997, 60, 195–198.
40. Rueda, A.; Zubía, E.; Ortega, M.J.; Carballo, J.L.; Salvá, J. New metabolites from the sponge Spongia agaricina. J. Nat. Prod. 1998, 61, 258–261.
41. Rosa, S.D.; Giulio, A.D.; Crisping, A.; Iodice, C.; Tommonaro, G. New 9,11-secosterol from the tyrrhenian sponge Fasciospongia Cavernosa. Nat. Prod. Lett. 1999, 13, 15–20.
42. Enwall, E.L.; van der Helm, D.; Hsu, I.N.; Pattabhiraman, T.; Schmitz, F.J.; Spraggins, R.L.; Weinheimer, A.J. Crystal structure and absolute configuration of two cyclopropane containing marine steroids. J. Chem. Soc. Chem. Comm. 1972, 215–216.
43. Schultz, L.W.; Clardy J.; Lessinger, L. 9,11-secogorgost-5-en-9-one-3,11-diol, a marine steroid from the sea whip Pseudopterogorgia hummelinkii. Acta Crystallogr. Sect. C 1995, C51.
44. Cheng, S.-Y.; Chen, H.-P.; Wang, S.-K.; Duh, C.-Y. Three new 9,11 secosterols from the Formosan soft coral Sinularia leptoclados. Bull. Chem. Soc. Jpn. 2011, 84, 648–652.
45. Kazlauskas, R.; Murphy, P.; Ravi, B.; Sanders, R.; Wells, R. Spermidine derivatives and 9,11-secosteroids from a soft coral Sinularia sp. Aust. J. Chem. 1982, 35, 69–75.
46. Su, J.-H.; Tseng, Y.-J.; Huang, H.-H.; Ahmed, A. F.; Lu, C.-K.; Wu, Y.-C.; Sheu, J.-H. 9,11-Secosterols from the Soft Corals Sinularia lochmodes and Sinularia leptoclados. J. Nat. Prod. 2006, 69, 850–852.
47. Bonini, C.; Cooper, C.B.; Kazlauskas, R.; Wells, R.J.; Djerassi, C. Minor and trace sterols in marine invertebrates. 41. Structure and stereochemistry of naturally occurring 9,11-seco sterols. J. Org. Chem. 1983, 48, 2108–2111.
48. He, H.; Kulanthaivel, P.; Baker, B.J.; Kalter, K.; Darges, J.; Cofield, D.; Wolff, L.; Adams, L. New antiproliferative and antiinflammatory 9,11-secosterols from the gorgonian Pseudopterogorgia sp. Tetrahedron 1995, 51, 51–58.
49. Naz, S.; Kerr, R.G.; Narayanan, R. New antiproliferative epoxysecosterols from Pseudopterogorgia americana. Tetrahedron Lett. 2000, 41, 6035–6040.
50. Rodríguez, A.D.; Rivera, J.; Boulanger, A. New polyhydroxydinostane sterols from the Caribbean gorgonian octocoral Pseudopterogorgia americana. Tetrahedron Lett. 1998, 39, 7645–7648.
51. Kobayashi, M.; Kobayashi, K.; Ramana, K.V.; Rao, C.V.L.; Rao, D.V.; Rao, C.B. Marine sterols. Part 20. polyhydroxy sterols of the soft corals of the Andaman and Nicobar coasts. Part 4. Andamansterol and nicobarsterol, novel sterols with 3,9,11,21-tetrahydroxylated, and 11,21-epoxy-9,11-secosteroid skeletons, from a Sclerophytum sp. of soft coral. X-Ray molecular structure of andamansterol. J. Chem. Soc. Perkin Trans. I 1991, 493–497.
52. Dopeso, J.; Quiñoá, E.; Riguera, R.; Debitus, C.; Bergquist, P.R. Euryspongiols: Ten new highly hydroxylated 9,11-secosteroids with antihistaminic activity from the sponge Euryspongia sp. Stereochemistry and reduction. Tetrahedron 1994, 50, 3813–3828.
53. Morris, L.A.; Christie, E.M.; Jaspars, M.; van Ofwegen, L.P. A bioactive secosterol with an unusual A- and B-ring oxygenation pattern isolated from an Indonesian soft coral Lobophytum sp. J. Nat. Prod. 1998, 61, 538–541.
54. Capon, R. J.; Faulkner, D.J. Herbasterol, an ichthyotoxic 9,11-secosterol from the sponge Dysidea herbacea. J. Org. Chem. 1985, 50, 4771–4773.
55. Van Altena, I.A.; Butler, A.J.; Dunne, S.J. A new cyclized 9,11-Secosterol enol−ether from the Australian Sponge Euryspongia arenaria. J. Nat. Prod. 1999, 62, 1154–1157.
56. Li, H.; Matsunaga, S.; Fusetani, N. A new 9,11-secosterol, stellettasterol from a marine sponge Stelletta sp. Experientia 1994, 50, 771–773.
57. Anta, C.; González, N.; Rodríguez, J.; Jiménez, C. A New secosterol from the Indonesian octocoral Pachyclavularia violacea. J. Nat. Prod. 2002, 65, 1357–1359.
58. Rodríguez Brasco, M.F.; Genzano, G.N.; Palermo, J.A. New C-secosteroids from the gorgonian Tripalea clavaria. Steroids 2007, 72, 908–913.
59. Van Wyk, A.W.W.; Gray, C.A.; Whibley, C.E.; Osoniyi, O.; Hendricks, D.T.; Caira, M.R.; Davies-Coleman, M.T. Bioactive metabolites from the south African marine mollusk Trimusculus costatus. J. Nat. Prod. 2008, 71, 420–425.
60. Chen, B.-W.; Chang, S.-M.; Huang, C.-Y.; Su, J.-H.; Wen, Z.-H.; Wu, Y.-C.; Sheu, J.-H. Hirsutosterols A-G, polyoxygenated steroids from a Formosan soft coral Cladiella hirsuta. Org Biomol Chem. 2011, 9, 3272–3278.
61. Zhang, G.; Tang, X.; Cheng, C.; Gong, K.; Zhang, X.; Zhu, H.; Wu, R.; Li, P.; Li, G. Cytotoxic 9,11-secosteroids from the South China Sea gorgonian Subergorgia suberosa. Steroids 2013, 78, 845–850.
62. Chen, W.-T.; Liu, H.-L.; Yao, L.-G.; Guo, Y.-W. 9,11-Secosteroids and polyhydroxylated steroids from two South China Sea soft corals Sarcophyton trocheliophorum and Sinularia flexibilis. Steroids 2014, 92, 56–61.
63. Yang, I.; Choi, H.; Won, D. H.; Nam, S.-J. Heonjoong Kang, An antibacterial 9,11-Secosterol from a marine sponge Ircinia sp. Bull. Korean Chem. Soc. 2014, 35, 1970–1974.
64. Zhang, J.; Liu, L.-L.; Zhong, B.-L.; Liao, X.-J.; Xu, S.-H. 9,11-Secosteroids with cytotoxic activity from the South China Sea gorgonian coral Subergorgia suberosa. Steroids 2015, 98, 100–106.
65. Yang, I.; Choi, H.; Nam, S.-J.; Kang, H. A new 9,11-secosterol with a 1,4-quinone from a Korean marine sponge Ircinia sp. Arch. Pharm. Res. 2015, 1–5.
66. Kawamura, A.; Kita, M.; Kigoshi, H. Aplysiasecosterol A: a 9,11-secosteroid with an unprecedented tricyclic -diketone structure from the sea hare Aplysia kurodai. Angew. Chem. Int. Ed. 2015, 54, 7073–7076.
67. Tomono, Y.; Hirota, H.; Fusetani, N. Isogosterones A−D, antifouling 13,17-secosteroids from an octocoral Dendronephthya sp. J. Org. Chem. 1999, 64, 2272–2275.
68. Fabricius, K.; Alderslade, P. “Soft Corals and Seafans–a Comprehensive Guide to the Tropical Shallow-water Genera of the Central–West Pacific, the Indian Ocean and Red Sea.＂Australian Institute of Marine Science, and the Museum and Art Gallery of the Northern Territory. 2001, pp 154, 155, 224, 225.
69. Kuo, L.-M.; Kuo, C.-Y.; Lin, C.-Y.; Hung, M.-F.; Shen, J.-J.; Hwang, T.-L. Intracellular glutathione depletion by oridonin leads to apoptosis in hepatic stellate cells. Molecules 2014, 19, 3327–3344.
70. Boyum, A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of mononuclear cells by one centrifugation and of granulocytes by combining centrifugation and sedimentation at 1 G. Scand. J. Clin. Lab. Invest. 1968, 97, 77–89.
71. Jauregui, H.O., Hayner, N.T., Driscoll, J.L., Williamsholland, R., Lipsky, M.H., Galletti, P.M. Trypan blue dye uptake and lactate-dehydrogenase in adult-rat hepatocytes-freshly isolated cells, cell-suspensions, and primary monolayer-cultures. In vitro 1981, 17, 1100–1110.
72. Babior, B.M.; Kipnes, R.S.; Curnutte, J.T. Biological defense mechanisms-production by leukocytes of superoxide a potential bactericidal agent. J. Clin. Invest. 1973, 52, 741–744.
73. Yu, H.-P.; Hsieh, P.-W.; Chang, Y.-J.; Chung, P.-J.; Kuo, L.-M.; Hwang, T.-L. 2-(2-Fluorobenzamido)benzoate ethyl ester (EFB-1) inhibits superoxide production by human neutrophils and attenuates hemorrhagic shock-induced organ dysfunction in rats. Free Radic. Biol. Med. 2011, 50, 1737–1748.
74. Yang, S.-C.; Chung, P.-J.; Ho, C.-M.; Kuo, C.-Y.; Hung, M.-F.; Huang, Y.-T.; Chang, W.-Y.; Chang, Y.-W.; Chan, K.-H.; Hwang, T.-L. Propofol inhibits superoxide production, elastase release, and chemotaxis in formyl peptide-activated human neutrophils by blocking formyl peptide receptor 1. J. Immunol. 2013, 190, 6511–6519.
75. Sright, J.L.C.; McInnes, A.G.; Shimizu, S.; Smith, D.G.; Walter, J.A.; Idler, D.; Khalil, W. Identification of C-24 alkyl epimers of marine sterols by 13C nuclear magnetic resonance spectroscopy. Can. J. Chem. 1978, 56, 1898–1903.
76. Cao, F.; Shao, C.-L.; Chen, M.; Zhang, M.-Q.; Xu, K.-X.; Meng, H.; Wang, C.-Y. Antiviral C-25 Epimers of 26-Acetoxy Steroids from the South China Sea Gorgonian Echinogorgia rebekka J. Nat. Prod. 2014, 77, 1488–1493.
77. Coutts, L.D.; Geiss, W.B.; Gregg, B.T.; Helle, M.A.; King, C.-H. R.; Itov, Z.; Mateo, M. E.; Meckler, H.; Zettler M. W.; Knutson, J. C. A Stereospecific Synthesis of 24(S)-Hydroxyvitamin D2, a Prodrug for 1α,24(S)- Dihydroxyvitamin D2. Org. Process Res. Dev. 2002, 6, 246–255.
78. Carmely, S.; Kashman, Y. Isolation and structure elucidation of lobophytosterol, depresosterol and three other closely related sterols. Tetrahedron, 1981, 37, 2397–2403.
79. Shen, T.; Zhang, L.; Wang, Y.-Y.; Fan, P.-H.; Wang, X.-N.; Lin, Z.-M.; Lou, H.-X. Steroids from Commiphora mukul display antiproliferative effect against human prostate cancer PC3 cells via induction of apoptosis. Bioorg. Med. Chem. Lett. 2012, 22, 4801–4806.
80. Santafé, G.; Paz, V.; Rodríguez, J.; Jiménez, C. Novel cytotoxic oxygenated C29 sterols from the Colombian marine sponge Polymastia tenax. J. Nat. Prod. 2002, 65, 1161–1164.
81. Yen, W.-H.; Chen, W.-F.; Cheng, C.-H.; Dai, C.-F.; Lu, M.-C.; Su, J.-H.; Su, Y.-D.; Chen, Y.-H.; Chang, Y.-C.; Chen, Y.-H.; Sheu, J.-H.; Lin, C.-S.; Wen, Z.-H.; Sung, P.-J. A novel 5α,8α-epidioxysterol from the soft coral Sinularia gaweli. Molecules 2013, 18, 2895–2903.
82. Blanc, P.-A.; Djerassi, C. Isolation and strucrure elucidation of 22(S),23(S)-methylenecholesterol. Evidence for direct bioalkylation of 22-dehydrocholesterol. J. Am. Chem. Soc. 1980, 102, 7113–7114.
83. Kawagishi, H.; Katsumi, R.; Sazawa, T.; Mizuno, T.; Hagiwara, T.; Nakamura, T. Cytotoxic steroids from the mushroom Agaricus blazei. Phytochemistry, 1988, 27, 2777–2779.
84. Kobayashi, M.; Krishna, M.; Haribabu, B.; Anjaneyulu, V. Marine sterol. XXV. Isolation of 23-demethylgorgost-7-ene-3,5,6-triol and (24S)-ergostane- 3,5,6-pentol from soft corals of the Andaman and Nicobar coasts. Chem. Pharm. Bull. 1993, 41, 87–89.
85. Kobori, M.; Yoshida M.; Ohnishi-Kameyama, M.; Takei, T.; Shinmoto. 5,8-Epidioxy-22E-ergosta-6,9(11),22-trien-3-ol from an edible mushroom suppresses growth of HL60 leukemia and HT29 colon adenocarcinoma cells. Biol. Pharm. Bull. 2006, 29, 755–759.
86. Lee, J.-S.; Ma, C.-M.; Park D.-K.; Yoshimi, Y.; Hantanaka, M.; Hattori, M. Transformation of ergosterol peroxide to cytotoxic substances by rat intestinal bacteria. Biol. Pharm. Bull. 2008, 31, 949–954.
87. Sheu, J.-H.; Chang, K.-C.; Duh, C.-Y. A cytotoxic 5,8-epidioxysterol from a soft coral Sinularia species. J. Nat. Prod. 2000, 63, 149–151.
88. Doi, Y.; Ishibashi, M.; Yamaguchi, N.; Kobayshi, J. Isolation of apo-9′-fucoxanthinone from the cultured marine dinoflagellate Amphidinium sp. J. Nat. Prod. 1995, 58, 1097–1099.
89. Zhang, H.; Xiao, X.; Conte, M.M.; Khalil, Z.; Capon, R.J. Spiralisones A–D: acylphloroglucinol hemiketals from an Australian marine brown alga, Zonaria spiralis. Org. Biomol. Chem. 2012, 10, 9671–9676.
90. Huang, S.-X.; Yang, J.; Xiao, W.-L.; Zhu, Y.-L.; Li, R.-T.; Li, L.-M.; Pu, J.-X.; Li, X.; Li, S.-H.; Sun, H.-D. Three novel terpenoids from Schisandra pubescens var. pubinervis. Helv. Chim. Acta. 2006, 89, 1169–1175.
91. Mizushina, Y.; Takeuchi, T.; Kuriyama, I.; Takahashi, Y.; Maezawa, S.; Matsumoto, T.; Koiwai, O.; Sugawara, F.; Yoshida, H. DNA polymerase  specific inhibitors, a loliolide derivative and pubinernoid A, isolated from the brown algae Laminaria sp. Lett. Drug. Des. Discov. 2011, 8, 671–677.
92. Li, S.; Wang, P.; Deng, G.; Yuan, W.; Su, Z. Cytotoxic compounds from invasive giant salvinia (Salvinia molesta) against human tumor cells. Bioorg. Med. Chem. Lett. 2013, 23, 6682–6687.
93. Wang, L.; Wang, L.-N.; Zhao, Y.; Lou, H.-X.; Cheng, A.-X. Secondary metabolites from Marchantia paleacea calluses and their allelopathic effects on Arabidopsis seed growth. Nat. Prod. Res. 2013, 27, 274–276.
94. Wang, L.; Yang, J.; Chi, Y.-Q.; Ouyang, W.-B.; Zang, Z.; Huang, S.-X.; Cao, P.; Zhao, Y. A new myrsinol-type diterpene polyester from Euphorbia dracunculoides Lam. Nat. Prod. Res. 2015, 29, 1406–1413.
95. Kobayshi, M.; Kanda, F. Marine sterols. 18. Isolation and structure of four novel oxygenated sterols from a gorgonian coral Melithaea ocracea. J. Chem. Soc. Perkin. Trans. 1991, 5, 1177–1179.
96. Yaotta, Y.; Ebdo, M.; Tani, Y.; Machida, K.; Amemiya, K.; Furumura, K.; Kikuchi, M. Sterol constituents from seven mushrooms. Chem. Pharm. Bull. 1999, 47, 847–851.
97. Yue, J.-M.; Chen, S.-N.; Lin, Z.-W.; Sun, H.-D. Sterols from the fungus Lactarium volemus. Phytochemistry 2001, 56, 801–806.
98. Santafé, G.; Paz, V.; Rodriguez, J.; Jiménez, C. Novel cytotoxic oxygenated C29 sterols from the Colombian marine sponge Polymastia tenax. J. Nat. Prod. 2002 65, 801–806.
99. Youssef, D.T.A.; Badr, J.M.; Shaala, L.A.; Mohamed, G.A.; Bamanie, F.H. Ehrenasterol and biemnic acid; new bioactive compounds from the red sea sponge Biemna ehrenbergi. Phytochem. Lett. 2015, 12, 296–301.
100. Luo, X.; Li, F.; Shinde, P.B.; Hong, J.; Lee, C.-O.; Im, K.S.; Jung, J.H. 26,27-cyclosterols and other polyoxygenated sterols from a marine sponge Topsentia sp. J. Nat. Prod. 2006 69, 1760–1768.
101. Chen, H.-L.; Chiang, H.-C. Constituents of fruit bodies of Tramete orientalis. J. Chinese Chem. Soc. 1995, 42, 97–100.
102. Ishizuka, T.; Yaotta, Y.; Kikuchi, M. Sterol constituents from the fruit bodies of Grifola frondosa (Fr.) Chem. Pharm. Bull. 1997, 45, 1756–1760.
103. Xu. L.; Korade, Z.; Porter, N.A. Oxysterols from free radical chain oxidation of 7-dehydrocholesterol: product and mechanistic studies. J. Am. Chem. Soc. 2010, 132, 2222–2232.
104. Epifanio, R.de; Maia, L.F.; Pawlik, J.R.; Fenical. W. Antipredatory secosterols from the octocoral Pseudoperogorgia Americana. Mar. Ecol. Prog. Ser. 2007, 329, 307–310.
105. Withers, N.W.; Kokkle, W.C.M.C.; Fenical, W.; Djerassi, C. Sterol patterns of cultured zooxanthellae isolated from marine invertebrate: synthesis of gorgosterol and 23-demethylgorgosterol by aposymbiotic algae. Proc. Natl. Acad. Sci. USA. 1982, 79, 3764–3768.
106. Ciereszko L.S. Sterol and diterpenoid production by zooxanthellae in coral reefs: a review. Biol. Oceanogr. 1989, 6, 363–374.
107. Jagodzinska, B.; Trimmer, J.S.; Fenical, W.; Djerassi, C. Sterols in marine invertebrates. 49. Isolation and structure elucidation of eight new polyhydroxylated sterols from the soft coral Sinularia. J. Org. Chern. 1985, 50, 1435–1439.
108. Kitagawa, I.; M. Kobayashi, Z.C.; Kiyota, Y.; Ohnishi. M. Marine natural products. XV. Chemical constituents of an Okinawan soft coral of Xenia sp. (Xenidae). Chern. Pharm. Bull. 1986, 34, 4590-4596.
109. Kerr, R.G.; Rodriguez, L.C.; Kellman, J.A chemoenzymatic synthesis of 9(11)-secosteroids using an enzyme extract of the marine gorgonian Pseudopterogorgia americana. Tetrahedron Lett. 1996, 37, 8301-8304.