Supporting Information Galanthamine, Plicamine, and Secoplicamine Alkaloids from Zephyranthes candida and Their Anti-acetylcholinesterase and Anti-inflammatory Activities Guanqun Zhan, Junfei Zhou, Rong Liu, Tingting Liu, Guoli Guo, Jianping Wang, Ming Xiang, Yongbo Xue, Zengwei Luo, Yonghui Zhang, and Guangmin Yao * Hubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, People s Republic of China List of Supporting Information Table S1. 1 H NMR Data for Compounds 7 11 in Methanol-d 4 (400 MHz)... 6 Table S2. 13 C NMR Data for Compounds 7 11 in Methanol-d 4 (100 MHz)... 7 Table S3. 1 H NMR Data for Compounds 12 16 in Methanol-d 4 (400 MHz)... 8 Table S4. 13 C NMR Data for Compounds 12 16 in Methanol-d 4 (100 MHz)... 9 Table S5. AChE Inhibitory Activities of Alkaloids 1 24 a... 10 Table S6. Inhibitory Effects of Alkaloids 1 24 a on the LPS-Induced NO Production in RAW264.7 Mouse Macrophages and the Cell Survival Rate... 10 Extraction and Isolation....11 Figure S1. Comparison of ECD spectra for 1 6 and 17 in MeOH.... 13 Figure S2. Comparison of ECD spectra for 7 16 and 22 in MeOH.... 13 Figure S3. Data fitting curves of the AChE inhibitory activities of compounds 3, 12 14, and 18 21... 14 Figure S4. Data fitting curves of the inhibitory effects of compounds 10 12, 14, 16, and dexamethasone on the LPS-induced NO production in RAW264.7 mouse macrophages... 14 Figure S5. (+)-HR-ESI-MS spectrum of 1.... 15 Figure S6. UV spectrum of 1 in MeOH.... 15 Figure S7. ECD spectrum of 1 in MeOH.... 16 Figure S8. IR spectrum of 1.... 16 Figure S9. 1 H NMR (400 MHz) spectrum of 1 in CD 3 OD.... 17 Figure S10. 13 C NMR (100 MHz) spectrum of 1 in CD 3 OD.... 17 Figure S11. DEPT 135 spectrum of 1 in CD 3 OD.... 18 * To whom correspondence should be addressed. Tel: 86-27-83691784. Fax: 86-027-83692762. E-mail: gyap@mail.hust.edu.cn. 1
Figure S12. HSQC spectrum of 1 in CD 3 OD.... 18 Figure S13. HMBC spectrum of 1 in CD 3 OD.... 19 Figure S14. 1 H- 1 H COSY spectrum of 1 in CD 3 OD.... 19 Figure S15. NOESY spectrum of 1 in CD 3 OD.... 20 Figure S16. (+)-HR-ESI-MS spectrum of 2.... 20 Figure S17. UV spectrum of 2 in MeOH.... 21 Figure S18. ECD spectrum of 2 in MeOH.... 21 Figure S19. IR spectrum of 2.... 22 Figure S20. 1 H NMR (400 MHz) spectrum of 2 in CD 3 OD.... 22 Figure S21. 13 C NMR (00 MHz) spectrumof 2 in CD 3 OD.... 23 Figure S22. DEPT 135 spectrum of 2 in CD 3 OD.... 23 Figure S23. HSQC spectrum of 2 in CD 3 OD.... 24 Figure S24. HMBC spectrum of 2 in CD 3 OD.... 24 Figure S25. 1 H- 1 H COSY spectrum of 2 in CD 3 OD.... 25 Figure S26. NOESY spectrum of 2 in CD 3 OD.... 25 Figure S27. (+)-HR-ESI-MS spectrum of 3.... 26 Figure S28. UV spectrum of 3 in MeOH.... 26 Figure S29. CD spectrum of 3 in MeOH.... 27 Figure S30. IR spectrum of 3.... 27 Figure S31. 1 H NMR (400 MHz) spectrum of 3 in CD 3 OD.... 28 Figure S32. 13 C NMR (100 MHz) spectrum of 3 in CD 3 OD.... 28 Figure S33. DEPT 135 spectrum of 3 in CD 3 OD.... 29 Figure S34. HSQC spectrum of 3 in CD 3 OD.... 29 Figure S35.HMBC spectrum of 3 in CD 3 OD.... 30 Figure S36. 1 H- 1 H COSY spectrum of 3 in CD 3 OD.... 30 Figure S37. NOESY spectrum of 3 in CD 3 OD.... 31 Figure S38. (+)-HR-ESI-MS spectrum of 4.... 31 Figure S39. UV spectrum of 4 in MeOH.... 32 Figure S40. ECD spectrum of 4 in MeOH.... 32 Figure S41. IR spectrum of 4.... 33 Figure S42. 1 H NMR (400 MHz) spectrum of 4 in CD 3 OD.... 33 Figure S43. 13 C NMR (100 MHz) spectrumof 4 in CD 3 OD.... 34 Figure S44. DEPT 135 spectrum of 4 in CD 3 OD.... 34 Figure S45. HSQC spectrum of 4 in CD 3 OD.... 35 Figure S46. HMBC spectrum of 4 in CD 3 OD.... 35 Figure S47. 1 H- 1 H COSY spectrum of 4 in CD 3 OD.... 36 Figure S48. NOESY spectrum of 4 in CD 3 OD.... 36 Figure S49. (+)-HR-ESI-MS spectrum of 5.... 37 Figure S50. UV spectrum of 5 in MeOH.... 37 Figure S51. ECD spectrum of 5 in MeOH.... 38 Figure S52. IR spectrum of 5.... 38 Figure S53. 1 H NMR (400 MHz) spectrum of 5 in CD 3 OD.... 39 Figure S54. 13 C NMR (100 MHz) spectrum of 5 in CD 3 OD.... 39 Figure S55. DEPT 135 spectrum of 5 in CD 3 OD.... 40 Figure S56. HSQC spectrum of 5 in CD 3 OD.... 40 Figure S57.HMBC spectrum of 5 in CD 3 OD.... 41 Figure S58. 1 H- 1 H COSY spectrum of 5 in CD 3 OD.... 41 Figure S59. NOESY spectrum of 5 in CD 3 OD.... 42 Figure S60. (+)-HR-ESI-MS spectrum of 6.... 42 Figure S61. UV spectrum of 6 in MeOH.... 43 Figure S62. ECD spectrum of 6 in MeOH.... 43 Figure S63. IR spectrum of 6.... 44 2
Figure S64. 1 H NMR (400 MHz) spectrum of 6 in CD 3 OD.... 44 Figure S65. 13 C NMR (100 MHz) spectrum of 6 in CD 3 OD.... 45 Figure S66. DEPT 135 spectrum of 6 in CD 3 OD.... 45 Figure S67. HSQC spectrum of 6 in CD 3 OD.... 46 Figure S68. HMBC spectrum of 6 in CD 3 OD.... 46 Figure S69. 1 H- 1 H COSY spectrum of 6 in CD 3 OD.... 47 Figure S70. NOESY spectrum of 6 in CD 3 OD.... 47 Figure S71. (+)-HR-ESI-MS spectrum of 7.... 48 Figure S72. UV spectrum of 7 in MeOH.... 48 Figure S73. ECD spectrum of 7 in MeOH.... 49 Figure S74. IR spectrum of 7.... 49 Figure S75. 1 H NMR (400 MHz) spectrum of 7 in CD 3 OD.... 50 Figure S76. 13 C NMR (100 MHz) spectrum of 7 in CD 3 OD.... 50 Figure S77. DEPT 135 spectrum of 7 in CD 3 OD.... 51 Figure S78. HSQC spectrum of 7 in CD 3 OD.... 51 Figure S79. HMBC spectrum of 7 in CD 3 OD.... 52 Figure S80. 1 H- 1 H COSY spectrum of 7 in CD 3 OD.... 52 Figure S81. NOESY spectrum of 7 in CD 3 OD.... 53 Figure S82. (+)-HR-ESI-MS spectrum of 8.... 53 Figure S83. UV spectrum of 8 in MeOH.... 54 Figure S84. ECD spectrum of 8 in MeOH.... 54 Figure S85. IR spectrum of 8.... 55 Figure S86. 1 H NMR (400 MHz) spectrum of 8 in CD 3 OD.... 55 Figure S87. 13 C NMR (100 MHz) spectrum of 8 in CD 3 OD.... 56 Figure S88. DEPT 135 spectrum of 8 in CD 3 OD.... 56 Figure S89. HSQC spectrum of 8 in CD 3 OD.... 57 Figure S90. HMBC spectrum of 8 in CD 3 OD.... 57 Figure S91. 1 H- 1 H COSY spectrum of 8 in CD 3 OD.... 58 Figure S92. NOESY spectrum of 8 in CD 3 OD.... 58 Figure S93. (+)-HR-ESI-MS spectrum of 9.... 59 Figure S94. UV spectrum of 9 in MeOH.... 59 Figure S95. ECD spectrum of 9 in MeOH.... 60 Figure S96. IR spectrum of 9.... 60 Figure S97. 1 H NMR (400 MHz) spectrum of 9 in CD 3 OD.... 61 Figure S98. 13 C NMR (100 MHz) spectrum of 9 in CD 3 OD.... 61 Figure S99. DEPT 135 spectrum of 9 in CD 3 OD.... 62 Figure S100. HSQC spectrum of 9 in CD 3 OD.... 62 Figure S101. HMBC spectrum of 9 in CD 3 OD.... 63 Figure S102. 1 H- 1 H COSY spectrum of 9 in CD 3 OD.... 63 Figure S103. NOESY spectrum of 9 in CD 3 OD.... 64 Figure S104. (+)-HR-ESI-MS spectrum of 10.... 64 Figure S105. UV spectrum of 10 in MeOH.... 65 Figure S106. ECD spectrum of 10 in MeOH.... 65 Figure S107. IR spectrum of 10.... 66 Figure S108. 1 H NMR (400 MHz) spectrum of 10 in CD 3 OD... 66 Figure S109. 13 C NMR (100 MHz) spectrum of 10 in CD 3 OD.... 67 Figure S110. DEPT 135 spectrum of 10 in CD 3 OD.... 67 Figure S111. HSQC spectrum of 10 in CD 3 OD.... 68 Figure S112.HMBC spectrum of 10 in CD 3 OD.... 68 Figure S113. 1 H- 1 H COSY spectrum of 10 in CD 3 OD.... 69 Figure S114. NOESY spectrum of 10 in CD 3 OD.... 69 Figure S115. (+)-HR-ESI-MS spectrum of 11.... 70 3
Figure S116. UV spectrum of 11 in MeOH.... 70 Figure S117. ECD spectrum of 11 in MeOH.... 71 Figure S118. IR spectrum of 11.... 71 Figure S119. 1 H NMR (400 MHz) spectrum of 11 in CD 3 OD.... 72 Figure S120. 13 C NMR (100 MHz) spectrum of 11 in CD 3 OD.... 72 Figure S121. DEPT 135 spectrum of 11 in CD 3 OD.... 73 Figure S122. HSQC spectrum of 11 in CD 3 OD... 73 Figure S123. HMBC spectrum of 11 in CD 3 OD.... 74 Figure S124. 1 H- 1 H COSY spectrum of 11 in CD 3 OD.... 74 Figure S125. NOESY spectrum of 11 in CD 3 OD.... 75 Figure S126. (+)-HR-ESI-MS spectrum of 12.... 75 Figure S127. UV spectrum of 12 in MeOH.... 76 Figure S128. ECD spectrum of 12 in MeOH.... 76 Figure S129. IR spectrum of 12.... 77 Figure S130. 1 H NMR (400 MHz) spectrum of 12 in CD 3 OD... 77 Figure S131. 13 C NMR (100 MHz) spectrum of 12 in CD 3 OD.... 78 Figure S132. DEPT 135 spectrum of 12 in CD 3 OD.... 78 Figure S133. HSQC spectrum of 12 in CD 3 OD.... 79 Figure S134. HMBC spectrum of 12 in CD 3 OD.... 79 Figure S135. 1 H- 1 H COSY spectrum of 12 in CD 3 OD.... 80 Figure S136. NOESY spectrum of 12 in CD 3 OD.... 80 Figure S137. (+)-HR-ESI-MS spectrum of 13.... 81 Figure S138. UV spectrum of 13 in MeOH.... 81 Figure S139. ECD spectrum of 13 in MeOH.... 82 Figure S140. IR spectrum of 13.... 82 Figure S141. 1 H NMR (400 MHz) spectrum of 13 in CD 3 OD... 83 Figure S142. 13 C NMR (100 MHz) spectrum of 13 in CD 3 OD.... 83 Figure S143. DEPT 135 spectrum of 13 in CD 3 OD.... 84 Figure S144. HSQC spectrum of 13 in CD 3 OD.... 84 Figure S145. HMBC spectrum of 13 in CD 3 OD.... 85 Figure S146. 1 H- 1 H COSY spectrum of 13 in CD 3 OD.... 85 Figure S147. NOESY spectrum of 13 in CD 3 OD.... 86 Figure S148. (+)-HR-ESI-MS spectrum of 14.... 86 Figure S149. UV spectrum of 14 in MeOH... 87 Figure S150. ECD spectrum of 14 in MeOH.... 87 Figure S151. IR spectrum of 14.... 88 Figure S152. 1 H NMR (400 MHz) spectrum of 14 in CD 3 OD... 88 Figure S153. 13 C NMR (100 MHz) spectrum of 14 in CD 3 OD.... 89 Figure S154. DEPT 135 spectrum of 14 in CD 3 OD.... 89 Figure S155. HSQC spectrum of 14 in CD 3 OD.... 90 Figure S156.HMBC spectrum of 14 in CD 3 OD.... 90 Figure S157. 1 H- 1 H COSY spectrum of 14 in CD 3 OD.... 91 Figure S158. NOESY spectrum of 14 in CD 3 OD.... 91 Figure S159. (+)-HR-ESI-MS spectrum of 15.... 92 Figure S160. UV spectrum of 15 in MeOH.... 92 Figure S161. ECD spectrum of 15 in MeOH.... 93 Figure S162. IR spectrum of 15.... 93 Figure S163. 1 H NMR (400 MHz) spectrum of 15 in CD 3 OD... 94 Figure S164. 13 C NMR (100 MHz) spectrum of 15 in CD 3 OD.... 94 Figure S165. DEPT 135 spectrum of 15 in CD 3 OD.... 95 Figure S166. HSQC spectrum of 15 in CD 3 OD.... 95 Figure S167. HMBC spectrum of 15 in CD 3 OD.... 96 4
Figure S168. 1 H- 1 H COSY spectrum of 15 in CD 3 OD.... 96 Figure S169. NOESY spectrum of 15 in CD 3 OD.... 97 Figure S170. (+)-HR-ESI-MS spectrum of 16.... 97 Figure S171. UV spectrum of 16 in MeOH.... 98 Figure S172. ECD spectrum of 16 in MeOH.... 98 Figure S173. IR spectrum of 16.... 99 Figure S174. 1 H NMR (400 MHz) spectrum of 16 in CD 3 OD... 99 Figure S175. 13 C NMR (100 MHz) spectrum of 16 in CD 3 OD.... 100 Figure S176. DEPT 135 spectrum of 16 in CD 3 OD.... 100 Figure S177. HSQC spectrum of 16 in CD 3 OD.... 101 Figure S178.HMBC spectrum of 16 in CD 3 OD.... 101 Figure S179. 1 H- 1 H COSY spectrum of 16 in CD 3 OD.... 102 Figure S180. NOESY spectrum of 16 in CD 3 OD.... 102 5
Table S1. 1 H NMR Data for Compounds 7 11 in Methanol-d 4 (400 MHz) position 7 8 9 10 11 1 5.91, d (9.9) 5.92, d (9.8) 5.91, d (9.9) 5.95, d (9.9) 5.80, d (9.9) 2 6.13, dd (9.9, 5.1) 6.11, dd (9.8, 5.2) 6.13, dd (9.9, 5.1) 6.12, dd (9.9, 5.1) 6.07, dd (9.9, 5.2) 3 3.95, ddd (5.1, 3.9, 3.4) 3.95, ddd (5.2, 3.6, 3.3) 3.95, ddd, (5.1, 4.2, 3.6) 3.96, ddd, (5.1, 4.0, 3.2) 3.92, ddd (5.2, 3.6, 3.3) 4α 1.62, overlap 1.58, overlap 1.62, ddd, (13.4, 11.7, 1.62, ddd, (13.5, 11.6, 1.58, ddd, (13.5, 11.6, 3.6) 3.2) 3.3) 4β 2.44, ddd, (13.4, 4.6, 2.45, ddd, (13.4, 4.6, 3.6) 2.44, ddd, (13.4, 4.6, 4.2) 2.45, ddd, (13.5, 4.6, 2.41, ddd, (13.5, 4.5, 3.6) 3.9) 4.0) 4a 3.77, dd, (11.5, 4.6) 3.78, dd (11.5, 4.6) 3.77, dd, (11.7, 4.6) 3.78, dd, (11.6, 4.6) 3.76, dd, (11.6, 4.5) 6α 3.80, d, (15.3) 3.79, d, (15.2) 3.79, d, (15.3) 3.79, d, (15.3) 3.83, d, (15.3) 6β 3.66, d, (15.3) 3.60, d, (15.2) 3.65, d, (15.3) 3.69, d, (15.3) 3.73, d, (15.3) 7 6.52, s 6.50, s 6.52, s 6.52, s 6.51, s 10 6.75, s 6.74, s 6.76, s 6.75, s 6.74, s 11 12 3.72, s 3.65, s 3.71, s 3.74, s 3.72, s OCH 2 O 5.89, d, (0.9); 5.87, d, (0.9) 5.88, d, (0.9); 5.87, d, (0.9) 5.89, d, (0.9); 5.88, d, (0.9) 5.89, d, (0.9); 5.88, d, (0.9) 5.88, d, (1.0); 5.87, d, (1.0) OCH 3-3 3.48, s 3.48, s 3.48, s 3.48, s 3.47, s NCH 3-5 2.72, s 2.70, s 2.72, s 2.72, s 2.70, s 13 3.04, m 2.93, dd, (12.9, 6.2); 2.71, 2.99, m 3.08, t, (6.8) 3.26, m overlap 14 1.44, m 1.60, overlap 1.52, m 1.84, m 2.83, m 15 1.64, overlap 1.46, m; 1.12, m 1.32, overlap 2.34, t, (7.2) 16 0.916, d, (6.5) 0.91, t, (7.4) 1.32, overlap 7.21, overlap 17 0.914, d, (6.5) 0.89, d, (6.5) 1.32, overlap 7.21, overlap 18 0.90, t, (6.9) 7.14, m 19 7.21, overlap 20 7.21, overlap 6
Table S2. 13 C NMR Data for Compounds 7 11 in Methanol-d 4 (100 MHz) position 7 8 9 10 11 1 137.1 137.5 137.1 137.1 137.1 2 126.2 125.8 126.2 126.1 126.0 3 72.8 72.9 72.8 72.8 72.8 4 30.8 30.8 30.8 30.8 30.7 4a 63.5 63.4 63.5 63.5 63.5 6 51.5 51.7 51.5 51.1 51.4 6a 130.2 130.7 130.2 130.0 130.2 7 107.0 106.8 107.0 107.0 106.9 8 148.2 148.1 148.2 148.3 148.2 9 148.3 148.2 148.3 148.4 148.3 10 109.3 109.3 109.3 109.3 109.3 10a 132.1 132.1 132.1 132.0 132.0 10b 45.8 46.2 45.8 45.9 45.9 11 174.3 174.6 174.3 174.2 174.3 12 67.2 68.5 67.2 67.7 67.5 OCH 2 O 102.5 102.4 102.5 102.5 102.5 OCH 3-3 57.0 57.0 57.0 57.0 57.0 NCH 3-5 28.1 28.1 28.10 28.1 28.0 13 54.1 62.9 55.9 55.5 57.8 14 37.9 34.5 28.8 24.4 35.7 15 27.5 28.6 33.0 33.5 141.8 16 23.24 11.8 28.14 178.9 130.1 17 23.22 18.0 23.9 129.4 18 14.5 127.1 19 129.4 20 130.1 7
Table S3. 1 H NMR Data for Compounds 12 16 in Methanol-d 4 (400 MHz) position 12 13 14 15 16 1 5.83, d (9.9) 5.86, ddd (10.6, 2.0, 1.8) 5.82, ddd (10.5, 2.0, 5.67, d (9.9) 5.67, d (9.9) 1.8) 2 6.06, dd (9.9, 5.1) 5.77, br, d (10.6) 5.74, br d (10.5) 6.30, dd (9.9, 5.1) 6.30, dd (9.9, 5.1) 3 3.90, ddd (5.1, 4.0, 3.3) 3.78, m 3.77, m 4.08, ddd (5.1, 4.0, 2.3) 4.09, ddd, (5.1, 4.0, 2.2) 4α 1.57, ddd, (13.5, 11.6, 2.04, ddd, (14.5, 10.2, 2.04, ddd, (14.3, 10.1, 2.68, ddd, (13.3, 13.1, 2.70, ddd, (13.3, 13.0, 4.0) 3.3) 2.0) 2.1) 4.0) 4β 2.40, ddd, (13.5, 4.5, 4.0) 2.81, ddd, (14.5, 4.8, 2.8) 2.81, overlap 1.98, ddd, (13.3, 2.7, 2.3) 2.00, ddd, (13.3, 2.8, 2.2) 4a 3.75, dd, (11.6, 4.5) 3.94, overlap 3.94, overlap 3.91, dd, (13.1, 2.7) 3.92, dd, (13.0, 2.8) 6α 3.87, d, (15.3) 4.27, d, (15.9) 4.30, d, (15.8) 4.37, d, (16.8) 4.42, d, (16.8) 6β 3.77, d, (15.3) 3.46, d, (15.9) 3.66, d, (15.8) 4.29, d, (16.8) 4.26, d, (16.8) 7 6.53, s 6.63, s 6.65, s 6.64, s 6.70, s 10 6.74, s 6.76, s 6.76, s 6.69, s 6.70, s 11 7.14, s 7.18, s 12 3.78, s 3.14, s 3.26, s OCH 2 O 5.89, d, (1.0); 5.88, d, 5.90, d, (0.9); 5.89, d, 5.90, d, (1.0); 5.89, d, 5.99, d, (1.0); 5.98, d, 5.99, d, (0.9); 5.98, d, (0.9) (1.0) (0.9) (1.0) (1.0) OCH 3-3 3.47, s 3.43, s 3.42, s 3.47, s 3.46, s NCH 3-5 2.70, s 2.86, s 2.85, s 2.70, s 2.64, s 13a 3.40, overlap 3.93, overlap 3.88, overlap 3.05, s 3.75, ddd, (13.1, 10.0, 6.1) 13b 3.30, overlap 2.38, ddd, (12.3, 8.9, 4.5) 2.74, overlap 3.19, ddd, (13.1,10.0, 5.4) 14 2.99, t, (7.5) 1.51, m; 1.41, m 2.77, overlap 1.49, m 15 1.68, m 1.59, m 16 0.95, d, (6.5) 7.08, d, (8.5) 0.955, d, (6.4) 17 7.53, br d, (7.8) 0.94, d, (6.5) 6.70, d, (8.0) 0.950, d, (6.4) 18 6.96, ddd, (7.8, 7.1, 1.0) 19 7.05, ddd, (8.1, 7.1, 1.0) 7.08, d, (8.5) 20 7.30, br d, (8.1) 6.70, d, (8.0) 22 7.04, s 8
Table S4. 13 C NMR Data for Compounds 12 16 in Methanol-d 4 (100 MHz) position 12 13 14 15 16 1 137.1 132.0 132.1 134.4 134.6 2 126.1 127.7 127.6 129.9 129.7 3 72.8 73.0 73.0 74.6 74.6 4 30.7 29.4 29.4 30.0 30.1 4a 63.5 58.8 58.8 63.4 63.4 6 51.6 58.3 58.2 53.1 51.1 6a 130.2 129.4 129.3 126.1 126.3 7 107.0 108.3 108.3 105.9 106.0 8 148.2 147.9 147.9 149.4 149.4 9 148.3 148.3 148.4 149.7 149.7 10 109.3 105.4 105.5 107.5 107.5 10a 132.1 133.5 133.5 130.4 130.6 10b 45.8 46.9 46.9 53.7 54.1 11 174.4 173.8 173.7 165.3 165.3 12 67.4 70.2 69.7 169.1 168.8 OCH 2 O 102.5 102.8 102.6 103.1 103.1 OCH 3-3 57.0 56.6 56.6 57.3 57.3 NCH 3-5 28.1 27.7 27.8 27.8 28.3 13 56.8 52.8 56.4 36.0 48.0 14 25.2 36.7 33.2 36.5 15 114.3 27.5 132.8 27.5 16 129.0 23.7 130.9 23.1 17 119.5 22.9 116.2 22.8 18 119.6 156.6 19 122.3 116.2 20 112.3 130.9 21 138.2 22 123.4 9
Table S5. AChE Inhibitory Activities of Alkaloids 1 24 a compounds IC 50 (μm) R 2 compounds IC 50 (μm) R 2 1 > 200-13 57.26 ± 3.76 0.909 2 > 200-14 75.30 ± 1.01 0.904 3 168.7 ± 20.36 0.996 15 > 200-4 > 200-16 > 200-5 > 200-17 > 200-6 > 200-18 b 1.25 ± 0.11 0.991 7 > 200-19 0.48 ± 0.01 0.996 8 > 200-20 40.12 ± 4.06 0.949 9 > 200-21 4.79 ± 0.51 0.994 10 > 200-22 > 200-11 > 200-23 > 200-12 110.6 ± 21.02 0.995 24 > 200 - a IC 50 values are expressed as the mean ± SD (n =3). b Galanthamine (18) was used as a positive control. Table S6. Inhibitory Effects of Alkaloids 1 24 a on the LPS-Induced NO Production in RAW264.7 Mouse Macrophages and the Cell Survival Rate compounds IC 50 (μm) R 2 survival rates (%) of RAW 264.7 at 200 μm LPS (0 μg/ml) LPS (1 μg/ml) 10 18.77 ± 0.91 0.96 116 ± 2 102 ± 3 11 10.21 ± 0.43 0.97 104 ± 3 108 ± 2 12 18.05 ± 0.73 0.94 114 ± 2 105 ± 3 14 7.50 ± 0.23 0.98 114 ± 2 99 ± 1 16 23.55 ± 0.14 0.98 113 ± 2 102 ± 3 dexamethasone b 0.63 ± 0.03 0.97 91 ± 1 100 ± 2 a Data are expressed as the mean ± SD. Other alkaloids, IC 50 > 200 μm. b Dexamethasone was used a positive control. 10
Extraction and Isolation. The dried whole plants of Z. candida (50 kg) were extracted with 95% aqueous EtOH at room temperature (25 L 4, each for one week). Each filtrate was concentrated and combined to afford a crude residue (3.78 kg). The residue was suspended in H 2 O (6 L), adjusted to ph = 2 with 0.3 M HCl,and extracted with CHCl 3 (10 3 L). The aqueous phase was adjusted to ph = 10 using an ammonia solution, and extracted with CHCl 3 (4 3 L) to afford 40 g CHCl 3 extract. Finally, the aqueous phase was partitioned with n-buoh to get 215 g n-buoh extract. The CHCl 3 extract (40 g) was subjected to silica gel CC (CHCl 3 MeOH, from 1:0 to 1:1) to afford five fractions (Fr. 1 5). Fr.1 was fractionated on a silica gel CC eluted with petroleum ether acetone (from 15:1 to 2:1), and then separated by RP HPLC (MeOH H 2 O Et 2 NH, 69.8:30:0.2) to afford compounds 12 (3.9 mg, t R 44.2 min), 23 (2.3 mg, 24.4 min), and 14 (3.5 mg, 29.6 min). Fr.2 was subjected to silica gel CC (petroleum ether acetone, from 12:1 to 2:1) to give four subfractions (Fr. 2A Fr.2D). Using RP HPLC (MeOH H 2 O Et 2 NH, 72.8:27:0.2), 7 (11.4 mg, t R 15.9 min) and 11 (4.7 mg, t R 36.6 min) were purified from the subfraction Fr.2AA, while, 13 (3.0 mg, t R 47.0 min), 8 (5.6 mg, t R 52.7 min), and 9 (10.1 mg, t R 62.4 min) were isolated from the subfraction Fr.2AB. Compounds 16 (11 mg, t R 32.1 min), 22 (2.8 mg, t R 18.6 min), and 10 (5.7 mg, t R 21.2 min) were separated from subfractions Fr.2B, Fr.2C, and Fr.2D, respectively, by RP C 18 HPLC with a mobile phase of MeCN H 2 O Et 2 NH (47:53:0.2), MeOH H 2 O Et 2 NH (59.8:40:0.2), and MeOH H 2 O (60:40), respectively. Fr.3 was subjected to repeated Sephadex LH-20 CC (MeOH) and RP C 18 MPLC (MeOH H 2 O, from 30:70 to 50:50) to yield subfractions Fr.3A and Fr.3B. Fr.3A was separated by RP HPLC (MeCN H 2 O Et 2 NH, 21.8:78:0.2) to yield 20 (48.2 mg, t R 45 min) and 18 (57.8 mg, t R 50 min). Compounds 15 (2.8 mg, t R 28.1 min) and 24 (9.2 mg, t R 32.5 min) were isolated from Fr.3B by RP HPLC (MeCN H 2 O Et 2 NH, 29.8:70:0.2). Fr.4 was fractionated by a silica gel CC (CHCl 3 MeOH, 100:1, 50:1, 20:1) to give two subfractions Fr.4A and Fr.4B. Compound 17 (258 mg) was isolated from Fr.4A by re-crystallization. Purification of Fr.4B by RP C 18 HPLC (MeOH H 2 O Et 2 NH, 34.8:65:0.2) afforded compound 1 (5.8 mg, t R 39.5 min). Fr.5 was fractionated using a silica gel CC (CHCl 3 MeOH, 50:1, 10:1, 5:1), and then purified by HPLC over a Welch AQ C 18 column (MeCN H 2 O Et 2 NH, 7.8:92:0.2) to get 4 (3 mg, t R 22.3 min) and 5 (4 mg, t R 23.8 min). The n-buoh extract (215 g) was subjected to a silica gel CC (CHCl 3 MeOH, from 30:1 to 1:1) to afford four fractions (Fr. I IV). Fr. I was fractionated by an RP C 18 CC (MeOH H 2 O, 0:100, 20:80), then purified by Sephadex LH-20 (MeOH), silica gel CC (CHCl 3 MeOH, 20:1, 3:1), and finally by RP HPLC (MeOH H 2 O Et 2 NH, 29.8:70:0.2) to yield 19 (28.2 mg, t R 18.0 min) and 21 (38.7 mg, t R 20.0 min). Fr. II was separated by an RP C 18 CC (MeOH H 2 O, 0:100), and then purified by HPLC over a Welch AQ C 18 column (MeCN H 2 O Et 2 NH, 0.8:99:0.2) to get 3 (4.3 mg, t R 24.7 min). The MeOH-insoluble solid was filtered out from Fr. III, and identified to be lycorine by co-tlc analysis with the authentic sample. Then, the filtrate of Fr. III was concentrated and fractionated by silica gel CC (CHCl 3 MeOH, from 30:1 to 3:1), Sephadex LH-20 CC (MeOH), and RP HPLC (MeOH H 2 O Et 2 NH, 9.8:90:0.2) to achieve compound 2 (5.8 mg, t R 29.3 min). In a similar manner, compound 6 (2.3 mg, t R 53.5 min) was finally isolated from Fr. IV by RP 11
HPLC (MeOH H 2 O Et 2 NH, 25:74.8:0.2). 12
Figure S1. Comparison of ECD spectra for 1 6 and 17 in MeOH. Figure S2. Comparison of ECD spectra for 7 16 and 22 in MeOH. 13
Figure S3. Data fitting curves of the AChE inhibitory activities of compounds 3, 12 14, and 18 21 Figure S4. Data fitting curves of the inhibitory effects of compounds 10 12, 14, 16, and dexamethasone on the LPS-induced NO production in RAW264.7 mouse macrophages 14
zgq1-74-6 #28 RT: 0.48 AV: 1 NL: 1.23E6 T: FTMS + p ESI Full ms [50.00-1500.00] 306.1654 100 90 80 70 Relative Abundance 60 50 40 30 20 328.1470 10 0 304.1498 344.1206 330.1540 312.1546 320.1809 342.1053 346.1186 360.3567 379.2542 290 300 310 320 330 340 350 360 370 380 m/z Figure S5. (+)-HR-ESI-MS spectrum of 1. Figure S6. UV spectrum of 1 in MeOH. 15
Figure S7. ECD spectrum of 1 in MeOH. Transmittance [%] 20 30 40 50 60 3339.96 2935.68 1834.61 1592.04 1625.63 1666.01 1507.74 1445.60 1364.66 1179.27 1127.20 1281.15 1250.38 1197.59 1160.89 1071.77 1032.94 974.81 923.75 800.45 943.82 838.07 774.06 716.61 856.02 671.60 646.14 616.79 731.02 595.92 554.65 882.34 522.69 470.91 419.76 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S8. IR spectrum of 1. 16
Figure S9. 1 H NMR (400 MHz) spectrum of 1 in CD 3 OD. Figure S10. 13 C NMR (100 MHz) spectrum of 1 in CD 3 OD. 17
Figure S11. DEPT 135 spectrum of 1 in CD 3 OD. Figure S12. HSQC spectrum of 1 in CD 3 OD. 18
Figure S13. HMBC spectrum of 1 in CD 3 OD. Figure S14. 1 H- 1 H COSY spectrum of 1 in CD 3 OD. 19
Figure S15. NOESY spectrum of 1 in CD 3 OD. ZGQ2-36-12 #22 RT: 0.31 AV: 1 NL: 1.19E8 T: FTMS + p ESI Full ms [50.00-1500.00] 100 292.15393 90 80 70 Relative Abundance 60 50 40 30 20 293.15723 10 0 291.14169 292.44348 294.16068 295.16083 298.16187 299.16507 291 292 293 294 295 296 297 298 299 m/z Figure S16. (+)-HR-ESI-MS spectrum of 2. 20
Figure S17. UV spectrum of 2 in MeOH. Figure S18. ECD spectrum of 2 in MeOH. 21
E:\ 同济医学院 \ 姚广民 \20140113\ZGQ2-36-12.0 Transmittance [%] 70 75 80 85 90 95 100 105 3331.72 2939.14 1599.27 1625.16 1508.90 1453.01 1368.31 1295.68 1156.75 1026.79 1250.56 1209.35 1179.83 1092.15 777.02 637.18 956.59 1118.21 1060.98 846.47 806.41 716.01 672.68 477.86 986.05 925.24 869.83 740.42 597.17 557.34 510.69 425.06 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S19. IR spectrum of 2. Figure S20. 1 H NMR (400 MHz) spectrum of 2 in CD 3 OD. 22
Figure S21. 13 C NMR (00 MHz) spectrumof 2 in CD 3 OD. Figure S22. DEPT 135 spectrum of 2 in CD 3 OD. 23
Figure S23. HSQC spectrum of 2 in CD 3 OD. Figure S24. HMBC spectrum of 2 in CD 3 OD. 24
Figure S25. 1 H- 1 H COSY spectrum of 2 in CD 3 OD. Figure S26. NOESY spectrum of 2 in CD 3 OD. 25
ZGQ2-41-1 #23 RT: 0.32 AV: 1 NL: 6.31E7 T: FTMS + p ESI Full ms [50.00-1500.00] 290.13821 100 90 80 70 Relative Abundance 60 50 40 30 20 10 0 291.14151 289.29245 288.28943 292.15375 293.15710 296.14618 297.23920 298.78873 288 290 292 294 296 298 300 m/z Figure S27. (+)-HR-ESI-MS spectrum of 3. Figure S28. UV spectrum of 3 in MeOH. 26
Figure S29. CD spectrum of 3 in MeOH. E:\ 同济医学院 \ 姚广民 \20131216IR\ZGQ2-41-1.0 3356.26 2919.53 2850.67 Transmittance [%] 55 60 65 70 75 80 85 1736.62 1598.97 1417.61 1508.54 1453.56 1384.07 1296.54 1204.82 1165.13 1032.10 1085.51 985.32 957.38 939.97 869.54 807.87 778.11 759.01 685.88 641.87 594.74 551.81 521.65 425.18 3500 3000 2500 2000 1500 1000 500 Wavenumber cm-1 Figure S30. IR spectrum of 3. 27
Figure S31. 1 H NMR (400 MHz) spectrum of 3 in CD 3 OD. Figure S32. 13 C NMR (100 MHz) spectrum of 3 in CD 3 OD. 28
Figure S33. DEPT 135 spectrum of 3 in CD 3 OD. Figure S34. HSQC spectrum of 3 in CD 3 OD. 29
Figure S35.HMBC spectrum of 3 in CD 3 OD. Figure S36. 1 H- 1 H COSY spectrum of 3 in CD 3 OD. 30
Figure S37. NOESY spectrum of 3 in CD 3 OD. zgq2-60-4 #22 RT: 0.32 AV: 1 NL: 2.30E7 T: FTMS + p ESI Full ms [50.00-1500.00] 100 322.1646 90 80 70 Relative Abundance 60 50 40 30 20 323.1679 10 0 320.1489 324.1716 325.1717 328.1725 331.1879 320 322 324 326 328 330 332 m/z Figure S38. (+)-HR-ESI-MS spectrum of 4. 31
Figure S39. UV spectrum of 4 in MeOH. Figure S40. ECD spectrum of 4 in MeOH. 32
E:\ 同济医学院 \ 姚广民 \20140605\ZGQ2-60-4.0 3324.65 Transmittance [%] 45 50 55 60 65 70 75 80 2940.40 1592.88 1624.82 1667.10 1511.71 1446.79 1409.87 1355.55 1286.08 1167.17 1098.26 1253.14 1203.43 1129.33 1030.30 780.48 981.48 924.41 811.46 645.31 947.11 716.16 671.79 900.56 856.94 836.80 740.95 566.46 453.07 481.98 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S41. IR spectrum of 4. Figure S42. 1 H NMR (400 MHz) spectrum of 4 in CD 3 OD. 33
Figure S43. 13 C NMR (100 MHz) spectrumof 4 in CD 3 OD. Figure S44. DEPT 135 spectrum of 4 in CD 3 OD. 34
Figure S45. HSQC spectrum of 4 in CD 3 OD. Figure S46. HMBC spectrum of 4 in CD 3 OD. 35
Figure S47. 1 H- 1 H COSY spectrum of 4 in CD 3 OD. Figure S48. NOESY spectrum of 4 in CD 3 OD. 36
zgq2-60-3 #20 RT: 0.31 AV: 1 NL: 2.80E7 T: FTMS + p ESI Full ms [50.00-1500.00] 320.1490 100 90 80 70 Relative Abundance 60 50 40 30 20 321.1523 10 0 322.1562 317.1147 323.1558 325.7648 317 318 319 320 321 322 323 324 325 326 327 328 m/z Figure S49. (+)-HR-ESI-MS spectrum of 5. Figure S50. UV spectrum of 5 in MeOH. 37
Figure S51. ECD spectrum of 5 in MeOH. E:\ 同济医学院 \ 姚广民 \20140605\ZGQ2-60-3.0 1284.88 Transmittance [%] 40 50 60 70 80 90 100 3310.45 2932.01 2842.99 1625.59 1593.47 1511.84 1445.91 1413.47 1356.26 1102.56 1257.10 1216.32 1171.95 1132.57 1047.09 1066.52 982.54 849.53 940.61 879.93 812.64 769.53 903.30 562.43 447.14 685.41 645.54 600.68 522.31 478.48 416.17 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S52. IR spectrum of 5. 38
Figure S53. 1 H NMR (400 MHz) spectrum of 5 in CD 3 OD. Figure S54. 13 C NMR (100 MHz) spectrum of 5 in CD 3 OD. 39
Figure S55. DEPT 135 spectrum of 5 in CD 3 OD. Figure S56. HSQC spectrum of 5 in CD 3 OD. 40
Figure S57.HMBC spectrum of 5 in CD 3 OD. Figure S58. 1 H- 1 H COSY spectrum of 5 in CD 3 OD. 41
Figure S59. NOESY spectrum of 5 in CD 3 OD. zgq2-15-6 #25 RT: 0.43 AV: 1 NL: 3.99E5 T: FTMS + p ESI Full ms [50.00-1500.00] 100 320.1446 90 80 70 Relative Abundance 60 50 40 30 342.1261 20 10 0 321.1479 322.1602 343.1295 327.1291 331.3043 336.1392 344.1417 350.1906 352.6205 320 325 330 335 340 345 350 m/z Figure S60. (+)-HR-ESI-MS spectrum of 6. 42
Figure S61. UV spectrum of 6 in MeOH. Figure S62. ECD spectrum of 6 in MeOH. 43
E:\ 同济医学院 \ 姚广民 \20131216IR\ZGQ2-15-6-2.0 Transmittance [%] 75 80 85 90 3328.92 2919.91 2850.92 1734.17 1508.14 1449.58 1591.19 1662.55 1653.39 1646.40 1626.04 1541.19 1559.30 1362.78 1384.98 1281.59 1212.38 1165.27 1077.96 1095.01 1033.87 945.06 972.51 801.78 639.13 886.05 832.90 773.09 733.68 654.86 678.22 588.44 548.44 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber cm-1 Figure S63. IR spectrum of 6. Figure S64. 1 H NMR (400 MHz) spectrum of 6 in CD 3 OD. 44
Figure S65. 13 C NMR (100 MHz) spectrum of 6 in CD 3 OD. Figure S66. DEPT 135 spectrum of 6 in CD 3 OD. 45
Figure S67. HSQC spectrum of 6 in CD 3 OD. Figure S68. HMBC spectrum of 6 in CD 3 OD. 46
Figure S69. 1 H- 1 H COSY spectrum of 6 in CD 3 OD. Figure S70. NOESY spectrum of 6 in CD 3 OD. 47
zgq2-12-6 #27 RT: 0.46 AV: 1 NL: 9.76E5 T: FTMS + p ESI Full ms [50.00-1500.00] 397.2061 100 90 80 399.2216 Relative Abundance 70 60 50 40 30 20 398.2097 399.6225 400.2251 10 0 401.2284 402.5059 405.2280 397 398 399 400 401 402 403 404 405 406 407 m/z Figure S71. (+)-HR-ESI-MS spectrum of 7. Figure S72. UV spectrum of 7 in MeOH. 48
Figure S73. ECD spectrum of 7 in MeOH. E:\ 同济医学院 \ 姚广民 \20140113\ZGQ2-21-4-1.0 2951.91 2868.86 1689.26 Transmittance [%] 55 60 65 70 75 80 85 3028.26 2819.81 1484.30 1503.90 1433.76 1386.39 1327.54 1273.31 1238.44 1190.54 1163.18 1101.55 1119.70 1082.47 1039.37 936.67 960.07 838.58 718.97 873.25 789.49 744.91 600.71 551.21 472.76 421.26 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S74. IR spectrum of 7. 49
Figure S75. 1 H NMR (400 MHz) spectrum of 7 in CD 3 OD. Figure S76. 13 C NMR (100 MHz) spectrum of 7 in CD 3 OD. 50
Figure S77. DEPT 135 spectrum of 7 in CD 3 OD. Figure S78. HSQC spectrum of 7 in CD 3 OD. 51
Figure S79. HMBC spectrum of 7 in CD 3 OD. Figure S80. 1 H- 1 H COSY spectrum of 7 in CD 3 OD. 52
zgq2-21-6 #24 RT: 0.41 AV: 1 NL: 4.51E5 T: FTMS + p ESI Full ms [50.00-1500.00] 399.2217 100 397.2061 90 80 Figure S81. NOESY spectrum of 7 in CD 3 OD. 70 Relative Abundance 60 50 40 30 20 400.2250 421.2032 10 0 401.2287 413.2370 411.2212 405.2296 415.2162 395 400 405 410 415 420 m/z 422.2066 Figure S82. (+)-HR-ESI-MS spectrum of 8. 53
Figure S83. UV spectrum of 8 in MeOH. Figure S84. ECD spectrum of 8 in MeOH. 54
E:\ 同济医学院 \ 姚广民 \20131216IR\ZGQ2-21-6-1.0 Transmittance [%] 50 55 60 65 70 75 80 85 3448.04 2925.49 2820.22 3028.65 2955.41 2874.05 1687.79 1576.53 1483.04 1431.97 1235.96 1503.19 1458.03 1386.87 1162.23 1100.94 1080.67 1037.47 1312.94 1331.16 1274.78 1188.54 1123.35 960.32 933.86 872.59 838.41 787.16 767.02 744.19 719.51 654.04 590.44 684.26 617.99 549.07 438.97 468.11 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S85. IR spectrum of 8. Figure S86. 1 H NMR (400 MHz) spectrum of 8 in CD 3 OD. 55
Figure S87. 13 C NMR (100 MHz) spectrum of 8 in CD 3 OD. Figure S88. DEPT 135 spectrum of 8 in CD 3 OD. 56
Figure S89. HSQC spectrum of 8 in CD 3 OD. Figure S90. HMBC spectrum of 8 in CD 3 OD. 57
Figure S91. 1 H- 1 H COSY spectrum of 8 in CD 3 OD. Figure S92. NOESY spectrum of 8 in CD 3 OD. 58
zgq2-21-7 #28 RT: 0.48 AV: 1 NL: 6.82E5 T: FTMS + p ESI Full ms [50.00-1500.00] 411.2214 100 413.2369 90 80 Relative Abundance 70 60 50 40 30 20 412.2248 414.2403 10 0 412.9467 415.2437 416.2475 411 412 413 414 415 416 417 m/z Figure S93. (+)-HR-ESI-MS spectrum of 9. Figure S94. UV spectrum of 9 in MeOH. 59
E:\ 同济医学院 \ 姚广民 \20131217\ZGQ2-21-7.0 Figure S95. ECD spectrum of 9 in MeOH. Transmittance [%] 20 30 40 50 60 70 3455.24 3028.94 2926.79 2856.54 2820.16 1689.10 1483.36 1239.26 1082.21 1037.86 1502.99 1432.25 1387.26 1160.00 1101.01 1330.86 1272.49 1187.91 1123.37 959.01 934.81 872.87 838.17 787.46 744.17 768.34 718.29 589.86 550.22 473.26 420.79 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S96. IR spectrum of 9. 60
Figure S97. 1 H NMR (400 MHz) spectrum of 9 in CD 3 OD. Figure S98. 13 C NMR (100 MHz) spectrum of 9 in CD 3 OD. 61
Figure S99. DEPT 135 spectrum of 9 in CD 3 OD. Figure S100. HSQC spectrum of 9 in CD 3 OD. 62
Figure S101. HMBC spectrum of 9 in CD 3 OD. Figure S102. 1 H- 1 H COSY spectrum of 9 in CD 3 OD. 63
Intens. Figure S103. NOESY spectrum of 9 in CD 3 OD. 1+ 415.1858 +MS, 0.1 0.2min #3 11 8000 1+ 437.1690 6000 4000 1+ 327.1353 1+ 398.2411 2000 0 274.2720 1+ 349.1163 451.1555 275 300 325 350 375 400 425 450 475 500 m/z Figure S104. (+)-HR-ESI-MS spectrum of 10. 64
Figure S105. UV spectrum of 10 in MeOH. Figure S106. ECD spectrum of 10 in MeOH. 65
E:\ 同济医学院 \ 姚广民 \20131217\ZGQ2-26-1.0 1688.08 1484.90 Transmittance [%] 30 40 50 60 70 80 3386.04 2929.07 2823.64 1598.76 1503.60 1436.76 1393.00 1332.21 1240.49 1189.69 1087.66 1037.27 1125.17 932.57 838.49 958.78 874.81 720.44 983.96 788.71 745.36 623.36 476.77 3500 3000 2500 2000 1500 1000 500 Wavenumber cm-1 Figure S107. IR spectrum of 10. Figure S108. 1 H NMR (400 MHz) spectrum of 10 in CD 3 OD. 66
Figure S109. 13 C NMR (100 MHz) spectrum of 10 in CD 3 OD. Figure S110. DEPT 135 spectrum of 10 in CD 3 OD. 67
Figure S111. HSQC spectrum of 10 in CD 3 OD. Figure S112.HMBC spectrum of 10 in CD 3 OD. 68
Figure S113. 1 H- 1 H COSY spectrum of 10 in CD 3 OD. Figure S114. NOESY spectrum of 10 in CD 3 OD. 69
zgq2-12-7 #28 RT: 0.48 AV: 1 NL: 2.02E6 T: FTMS + p ESI Full ms [50.00-1500.00] 433.2054 100 90 80 70 Relative Abundance 60 50 40 30 434.2089 20 10 0 434.1039 433.0808 433.3273 435.2120 436.2153 433 434 435 436 437 438 m/z Figure S115. (+)-HR-ESI-MS spectrum of 11. 436.7286 Figure S116. UV spectrum of 11 in MeOH. 70
Figure S117. ECD spectrum of 11 in MeOH. E:\ 同济医学院 \ 姚广民 \20131216IR\ZGQ2-12-7.0 1689.09 1483.54 Transmittance [%] 60 70 80 90 100 3462.50 3026.21 2892.61 2929.61 2819.86 1602.50 1432.86 1238.51 1085.70 1037.94 1501.60 1454.09 1387.85 1313.92 1330.94 1273.70 1188.19 1154.86 1124.61 958.75 934.17 873.24 838.61 789.07 746.66 700.41 623.96 571.87 472.84 506.33 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S118. IR spectrum of 11. 71
Figure S119. 1 H NMR (400 MHz) spectrum of 11 in CD 3 OD. Figure S120. 13 C NMR (100 MHz) spectrum of 11 in CD 3 OD. 72
Figure S121. DEPT 135 spectrum of 11 in CD 3 OD. Figure S122. HSQC spectrum of 11 in CD 3 OD. 73
Figure S123. HMBC spectrum of 11 in CD 3 OD. Figure S124. 1 H- 1 H COSY spectrum of 11 in CD 3 OD. 74
Figure S125. NOESY spectrum of 11 in CD 3 OD. ZGQ2-50-1 #26 RT: 0.36 AV: 1 NL: 1.32E8 T: FTMS + p ESI Full ms [50.00-1500.00] 100 472.2180 90 80 70 Relative Abundance 60 50 40 30 20 472.2665 473.2216 10 0 473.2699 474.2246 472.5 473.0 473.5 474.0 474.5 m/z Figure S126. (+)-HR-ESI-MS spectrum of 12. 75
Figure S127. UV spectrum of 12 in MeOH. Figure S128. ECD spectrum of 12 in MeOH. 76
E:\ 同济医学院 \ 姚广民 \20131217\ZGQ2-50-1.0 2918.25 Transmittance [%] 20 30 40 50 60 70 80 90 1674.40 1455.16 1481.60 1501.63 1432.30 1386.78 1235.83 1187.77 1084.70 1036.88 1122.56 932.22 872.81 837.84 788.21 742.52 477.28 425.76 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S129. IR spectrum of 12. Figure S130. 1 H NMR (400 MHz) spectrum of 12 in CD 3 OD. 77
Figure S131. 13 C NMR (100 MHz) spectrum of 12 in CD 3 OD. Figure S132. DEPT 135 spectrum of 12 in CD 3 OD. 78
Figure S133. HSQC spectrum of 12 in CD 3 OD. Figure S134. HMBC spectrum of 12 in CD 3 OD. 79
Figure S135. 1 H- 1 H COSY spectrum of 12 in CD 3 OD. Figure S136. NOESY spectrum of 12 in CD 3 OD. 80
zgq2-21-5 #17 RT: 0.28 AV: 1 NL: 3.89E3 T: FTMS + p ESI Full ms [50.00-1500.00] 415.2160 100 90 80 70 Relative Abundance 60 50 40 30 416.2193 20 417.2317 10 0 415.0 415.5 416.0 416.5 417.0 417.5 m/z Figure S137. (+)-HR-ESI-MS spectrum of 13. Figure S138. UV spectrum of 13 in MeOH. 81
Figure S139. ECD spectrum of 13 in MeOH. E:\ 同济医学院 \ 姚广民 \20131217\ZGQ2-21-5.0 Transmittance [%] 10 20 30 40 50 60 70 3383.55 2926.52 2821.05 2952.00 2868.81 3037.13 1696.71 1483.29 1503.36 1367.64 1388.73 1239.43 1295.09 1194.79 1077.73 1093.83 1036.72 933.02 967.36 802.78 838.75 857.60 697.68 726.16 671.42 632.06 588.57 457.04 482.59 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S140. IR spectrum of 13. 82
Figure S141. 1 H NMR (400 MHz) spectrum of 13 in CD 3 OD. Figure S142. 13 C NMR (100 MHz) spectrum of 13 in CD 3 OD. 83
Figure S143. DEPT 135 spectrum of 13 in CD 3 OD. Figure S144. HSQC spectrum of 13 in CD 3 OD. 84
Figure S145. HMBC spectrum of 13 in CD 3 OD. Figure S146. 1 H- 1 H COSY spectrum of 13 in CD 3 OD. 85
ZGQ2-49-6 #27 RT: 0.38 AV: 1 NL: 2.73E5 T: FTMS + p ESI Full ms [50.00-1500.00] 465.1973 100 90 Figure S147. NOESY spectrum of 13 in CD 3 OD. 80 70 Relative Abundance 60 50 40 30 466.2010 467.2114 20 10 0 465.5 466.0 466.5 467.0 m/z Figure S148. (+)-HR-ESI-MS spectrum of 14. 86
Figure S149. UV spectrum of 14 in MeOH Figure S150. ECD spectrum of 14 in MeOH. 87
E:\ 同济医学院 \ 姚广民 \20131217\ZGQ2-49-6.0 3332.75 2924.44 Transmittance [%] 20 30 40 50 60 70 80 90 1672.28 1612.21 1513.52 1482.07 1448.83 1391.43 1367.33 1239.20 1296.41 1081.32 1036.04 931.98 817.66 726.53 564.61 633.73 590.38 529.10 3500 3000 2500 2000 1500 1000 500 Wavenumber cm-1 Figure S151. IR spectrum of 14. Figure S152. 1 H NMR (400 MHz) spectrum of 14 in CD 3 OD. 88
Figure S153. 13 C NMR (100 MHz) spectrum of 14 in CD 3 OD. Figure S154. DEPT 135 spectrum of 14 in CD 3 OD. 89
Figure S155. HSQC spectrum of 14 in CD 3 OD. Figure S156.HMBC spectrum of 14 in CD 3 OD. 90
Figure S157. 1 H- 1 H COSY spectrum of 14 in CD 3 OD. Figure S158. NOESY spectrum of 14 in CD 3 OD. 91
ZGQ2-45-2 #25 RT: 0.35 AV: 1 NL: 3.08E7 T: FTMS + p ESI Full ms [50.00-1500.00] 359.1599 100 90 80 70 Relative Abundance 60 50 40 30 20 360.1631 10 0 360.3229 361.1673 359.0 359.5 360.0 360.5 361.0 361.5 m/z Figure S159. (+)-HR-ESI-MS spectrum of 15. Figure S160. UV spectrum of 15 in MeOH. 92
E:\ 同济医学院 \ 姚广民 \20131217\ZGQ2-45-2-1.0 Figure S161. ECD spectrum of 15 in MeOH. 1644.57 1672.69 1485.34 Transmittance [%] 45 50 55 60 65 70 75 80 3475.80 2929.01 3030.35 2824.26 1504.51 1236.81 1405.51 1315.53 1262.32 1427.94 1339.62 1362.46 1090.28 1038.17 1111.56 1170.47 933.46 825.81 750.87 854.02 764.11 664.78 786.75 729.99 691.25 645.02 591.06 536.76 470.72 489.37 429.95 3500 3000 2500 2000 1500 Wavenumber cm-1 1000 500 Figure S162. IR spectrum of 15. 93
Figure S163. 1 H NMR (400 MHz) spectrum of 15 in CD 3 OD. Figure S164. 13 C NMR (100 MHz) spectrum of 15 in CD 3 OD. 94
Figure S165. DEPT 135 spectrum of 15 in CD 3 OD. Figure S166. HSQC spectrum of 15 in CD 3 OD. 95
Figure S167. HMBC spectrum of 15 in CD 3 OD. Figure S168. 1 H- 1 H COSY spectrum of 15 in CD 3 OD. 96
Figure S169. NOESY spectrum of 15 in CD 3 OD. ZGQ2-50-4 #25 RT: 0.35 AV: 1 NL: 4.41E7 T: FTMS + p ESI Full ms [50.00-1500.00] 100 415.2220 90 80 70 Relative Abundance 60 50 40 30 20 416.2253 10 0 417.2289 415.0 415.5 416.0 416.5 417.0 417.5 418.0 418.5 m/z Figure S170. (+)-HR-ESI-MS spectrum of 16. 97
Figure S171. UV spectrum of 16 in MeOH. Figure S172. ECD spectrum of 16 in MeOH. 98
E:\ 同济医学院 \ 姚广民 \20131216IR\ZGQ2-50-4-1.0 Transmittance [%] 70 75 80 85 90 95 100 3481.07 3028.79 2931.59 2955.59 2870.72 2821.58 1672.79 1482.05 1424.99 1403.93 1366.67 1338.73 1293.43 1234.86 1078.62 1037.31 1091.88 1060.71 1168.46 1119.23 980.70 935.09 852.77 808.72 727.90 867.49 823.97 468.76 761.29 664.76 785.77 689.24 629.04 646.14 589.55 540.01 485.21 502.77 3500 3000 2500 2000 1500 1000 500 Wavenumber cm-1 Figure S173. IR spectrum of 16. Figure S174. 1 H NMR (400 MHz) spectrum of 16 in CD 3 OD. 99
Figure S175. 13 C NMR (100 MHz) spectrum of 16 in CD 3 OD. Figure S176. DEPT 135 spectrum of 16 in CD 3 OD. 100
Figure S177. HSQC spectrum of 16 in CD 3 OD. Figure S178.HMBC spectrum of 16 in CD 3 OD. 101
Figure S179. 1 H- 1 H COSY spectrum of 16 in CD 3 OD. Figure S180. NOESY spectrum of 16 in CD 3 OD. 102