1. J. Nat. Prod. 2004, 67, 1455-1458 New Nardosinanes and 19-Oxygenated Ergosterols from the Soft Coral Nephthea armata Collected in Taiwan Ali A. H. El-Gamal, Shang-Kwei Wang, Chang-Feng Dai, and Chang-Yih Duh Department of Marine Resources, National Sun Yat-sen University, Kaohsiung, Taiwan, Department of Microbiology, Kaohsiung Medical University, Kaohsiung, Taiwan, & Institute of Oceanography, National Taiwan University, Taipei, Taiwan, Republic of China

2. Abstract Five new nardosinane sesquiterpenoids, armatins A-E (1-5),lemnal-1(10)-ene-2,12-dione (6) (a new natural product), and two new cytotoxic 19-oxygenated ergosterols, armatinols A and B (7 and 8), were isolated from the methylene chloride extract of the soft coral Nephthea armata, collected in Taiwan. The structures were elucidated by 1D and 2D NMR spectral analysis, and their cytotoxicity was determined against selected cancer cells.

3. Nephthea Soft Coral Photo

4. Aim of study Separation, structural elucidation and biological study of the bioactive substances found in the cytotoxic part of methylene chloride extract of the soft corals Nephthea armata grow in Taiwan

5. Elution by n-hex-EtOAc (3:7) afforded fractions containing Compounds 1, 3-5 and 8 Compound 1 was further purified by silica gel CC using MeOH-CH2Cl2 (12:88) Compound 8 was further purified by silica gel CC using (MeOH: CH2Cl2, 25:27) Compounds 3-5 were further purified b C18 HPLC CC using MeOH: H2O (67:33) Bodies of soft coral N. armata were freeze dried to give 1.75 Kg, extract with CH2Cl2, conc. to give 27 g Silica gel CC with n-hexane- EtOAc and MeOH-EtOAc respectively Elution by MeOH-EtOAc (5:95) afforded fractions containing Compounds 2,6 and 7 Compound 2 was further purified by silica gel CC with n-hex.-EtOAc (1:1) as a solvent system Compound 7 was further purified by silica gel CC using acetone –CH2Cl2 (2:8) Compound 6 was obtained by C18 HPLC column using MeOH- H2O (65:35)

6. Nardosinanes Sesquiterpenes and 19 oxygenated Ergosterols isolated from Nephthea armata *Armartnol A (7) exhibited cytotoxicity against A-549, HT-29, and P-388 cells with IC 50 value of 7.6, 6.5, and 6.1  M, respectively. **Armatinol B (8) showed cytotoxicity against P-388 and HT-29 cells with IC 50 values of 3.2 and 3.1  M, respectively. The IC 50 values of compounds 1-6 against P-388, HT-29, and A-549 were greater than 50  M.

7. 1H NMR of compound 1 (Armatin A) 12 1 2, 7 1314 15 9 3,8,6 9,11

8. HMBC of Compound 1 (Armatin A) 1 10 12 7 2,7 1 12 9 9’,11 3,8,6

9. 2D INADEQUATE NMR spectrum for Compound 1

10. 1 H NMR of Compound 2 (Aramatin B) 2,7 1 15 14 13 9,11 6

11. 13 C NMR and DEPT experiment of Compound 2 (Aramatin B) 10 1 12 7 2 6 5,11,3 8,9,4 14,15, 16

12. HSQC of Compound 2 (Armatin B)

13. HMBC of Compound 2 (Armatin B)

14. 1H NMR of Compound 3 (Armatin C) 1 12 2 7 OMe 1 3 14 15

15. HMBC of Compound 3(Armatin C ) 7 12 1 10 2 6 OMe 112 2 7 11 5 3

16. 1HNMR of compound 4 (ArmatinD)

17. 13C NMR and DEPT Experiment of Compound 4 (Armatin D) 1 12 10 7 2 6, OMe 5 11 3 8 9 4 14 15 13

18. HSQC of compound 4 (Armatin D

19. 1 H NMR of compound 5 (Armatin E) 1 12 7 OMe 9 15 13 14 3434 11 9 8 6 8 x

20. COSY of compound 5 (Armatin E)

21. 1 H NMR of compound 6 (lemnal -1(10)-ene-2,12-dione) 17 15 14 13 x

22. COSY of compound 6

23. HSQC of Compound 6 176 1 7 6 5 15 14 13 8 4 1115 H 13 14 78

24. Expanded HMBC of Compound 6 7 6 5 7 11 48 14 13 15 1 7 6 8 14 13

25. m/z 248 C 15 H 20 O 3 EIMS of Compound 6

26. HMBC correlations of Compounds 1-6 (Armatins A- E)

27. 1HNMR of compound 7 (Armatinol A)

28. 13CNMR and DEPT Expermints of compound 7 (Armationl A)

29. 1HNMR of Compound 8 (Armatinol B) 19 28 3 6 OAc 18 21 26 27

30. 13CNMR and DEPT Expermints of Compound 8 (Armatinol B) 28 24 3 19 6 5 18 14 17 9