S1 of S21 Supplementary Materials: Bioactive Stilbenes from Cortex Mori Radicis, and Their Neuroprotective and Analgesic Activities Mediated by mglur1 Ya-Nan Wang, Mao-Feng Liu, Wei-Zhen Hou, Rui-Ming Xu, Jie Gao, An-Qi Lu, Mei-Ping Xie, Lan Li, Jian-Jun Zhang, Ying Peng, Li-Li Ma, Xiao-Liang Wang, Jian-Gong Shi and Su-Juan Wang Supporting information Figure SI-1a. 1 HNMR spectra of compound 1 Figure SI-1b. 13 CNMR spectra of compound 1 Figure SI-1c. HMQC spectra of compound 1 Figure SI-1d. HMBC spectra of compound 1 Figure SI-1e. (+)ESIMS spectra of compound 1 Figure SI-1f. IR spectra of compound 1 Figure SI-1g. CD and UV spectrum of compound 1 Figure SI-1h. The absolute configuration of sugar in 1 detered by HPLC Figure SI-2a. 1 HNMR spectra of compound 2 Figure SI-2b. 13 CNMR spectra of compound 2 Figure SI-2c. HMQC spectra of compound 2 Figure SI-2d. HMBC spectra of compound 2 Figure SI-2e. (+)ESIMS spectra of compound 2 Figure SI-2f. IR spectra of compound 2 Figure SI-2g. CD and UV spectrum of compound 2 Figure SI-2h. The absolute configuration of sugar in 2 detered by HPLC Figure SI-3a. 1 HNMR spectra of compound 3 Figure SI-3b. 13 CNMR spectra of compound 3 Figure SI-3c. HMQC spectra of compound 3 Figure SI-3d. HMBC spectra of compound 3 Figure SI-3e. (+)ESIMS spectra of compound 3 Figure SI-3f. IR spectra of compound 3 Figure SI-3g. CD and UV spectrum of compound 3 Figure SI-3h. The absolute configuration of sugar in 3 detered by HPLC Figure SI-4a. 1 HNMR spectra of compound 4 Figure SI-4b. 13 CNMR spectra of compound 4 Figure SI-4c. HMQC spectra of compound 4 Figure SI-4d. HMBC spectra of compound 4 Figure SI-4e. (+)ESIMS spectra of compound 4 Figure SI-4f. IR spectra of compound 4 Figure SI-4g. CD and UV spectrum of compound 4 Figure SI-4h. The absolute configuration of sugar in 4 detered by HPLC Table SI-1. Optical rotation of aglycones 1a 4a, moracin O (5), and P (8) Calculated ECD of 1a Figure SI-5a. Calculated ECD spectra of 1a-C1 C3
S2 of S21 Table SI-2. The Boltzmann distribution of three conformations (C1 C3) of 1a Figure SI-5b. MO related with two cotton effects of 1a HPLC profile of Cortex Mori Radicis and their main components identified by HPLC-MS Figure SI-6. HPLC profile of Cortex Mori Radicis Table SI-3. The main components of Cortex Mori Radicis identified by HPLC-MS Figure SI-1a. 1 H NMR spectra of compound 1.
S3 of S21 Figure SI-1b. 13 C NMR spectra of compound 1. Figure SI-1c. HMQC spectra of compound 1.
S4 of S21 Figure SI-1d. HMBC spectra of compound 1. x1 6 2. +MS,. #2 481.2 1.5 1. 274.3 31.2 381.3.5. 353.3 318.4 41.4 437.2 191. 248.1 2 25 3 35 4 45 m/z Figure SI-1e. (+)ESIMS spectra of compound 1.
S5 of S21 Figure SI-1f. IR spectra of compound 1. Figure SI-1g. CD and UV spectrum of compound 1.
S6 of S21 8 6 4 2 4 3 2 1 2 15 1 5 DAD1 A, Sig=25,4 Ref=36,1 (H:\21412212.D) 16.65 17.93 1 2 3 DAD1 A, Sig=25,4 Ref=36,1 (H:\214122-1.D) 1 2 3 DAD1 A, Sig=25,4 Ref=36,1 (H:\214122-2.D) 17.273 19.176 15.846 16.482 17.696 18.651 1 2 3 19.8 22.49 Compd 1* 22.34 24.34 L-ara 27.367 D-ara 36.21 Figure SI-1h. HPLC of compound 1, D-arabinose, L-arabinose. Figure SI-2a. 1 H NMR spectra of compound 2.
S7 of S21 Figure SI-2b. 13 C NMR spectra of compound 2. Figure SI-2c. HMQC spectra of compound 2.
S8 of S21 Figure SI-2d. HMBC spectra of compound 2. Intens. x1 6 +MS,.2 #11 8 511.2 6 475.4 4 2 274.3 31.2 362.3 318.3 385.2 191. 418.8 15 2 25 3 35 4 45 5 m/z Figure SI-2e. (+)ESIMS spectra of compound 2.
S9 of S21 Figure SI-2f. IR spectra of compound 2. Figure SI-2g. CD and UV spectrum of compound 2.
S1 of S21 DAD1 A, Sig=25,4 Ref=6,1 (H:\42-121.D) 3 2.625 Compd 2* 2 1 1.133 16.6 16.956 17.552 25.974 27.829 34.999 35.129-1 2 15 1 2 3 DAD1 A, Sig=25,4 Ref=6,1 (H:\35-131.D) 2.575 D-glc 1 5 8 1.11 1 2 3 DAD1 A, Sig=25,4 Ref=6,1 (H:\36-141.D) 18.845 19.823 L-glc 27.789 35.62 38.785 6 4 2 1.116 16.136 17.561 2.568 23.951 24.72 27.825 34.93 34.98 1 2 3 Figure SI-2h. HPLC of compound 2, D-glucose and L-glucose. Figure SI-3a. 1 H NMR spectra of compound 3.
S11 of S21 Figure SI-3b. 13 C NMR spectra of compound 3. Figure SI-3c. HMQC spectra of compound 3.
S12 of S21 compound 3 SBP-34-2/3 Bruker AVIIIHD 6 2141218 {HMBC etgpl3nd} DMSO D:\\ DATA214 5 2 3 4 5 6 7 8 9 1 11 12 13 14 15 16 7.5 7. 6.5 6. 5.5 5. 4.5 4. f2 (ppm) 3.5 3. 2.5 2. 1.5 1. Figure SI-3d. HMBC spectra of compound 3. 5 x1 2.4 2.2 2 1.8 1.6 1.4 1.2 1.8.6.4.2 + Scan (6.319 ) 21412223.d Subtract 24 25 26 27 28 29 3 31 32 33 34 35 36 37 38 39 4 41 42 43 44 45 46 47 48 49 5 51 52 53 Counts (%) vs. Mass-to-Charge (m/z) Figure SI-3e. (+)ESIMS spectra of compound 3.
S13 of S21 Figure SI-3f. IR spectra of compound 3. Figure SI-3g. CD and UV spectrum of compound 3.
S14 of S21 6 4 2 15 125 1 75 5 25 1 8 6 4 2 DAD1 A, Sig=25,4 Ref=36,1 (H:\2141225.D) 17.425 1 2 3 DAD1 A, Sig=25,4 Ref=36,1 (H:\2141228.D) 15.365 17.997 1 2 3 DAD1 A, Sig=25,4 Ref=36,1 (H:\2141229.D) 15.348 16.33 17.113 1 2 3 18.2 19.356 19.656 19.647 22.377 Compd 3* 23.671 24.95 25.72 D-glc L-glc 24.73 26.112 25.943 29.297 34.134 35.217 38.319 Figure SI-3h. HPLC of compound 3, D-glucose and L-glucose. Figure SI-4a. 1 H NMR spectra of compound 4.
S15 of S21 Figure SI-4b. 13 C NMR spectra of compound 4. Figure SI-4c. HMQC spectra of compound 4.
S16 of S21 compound 4 SBP-31-1/3 Bruker AVIIIHD 6 2141218 {HMBC etgpl3nd} DMSO D:\\ DATA214 48 1 2 3 4 5 6 7 8 9 1 11 12 13 14 15 16 7.5 7. 6.5 6. 5.5 5. 4.5 f2 (ppm) 4. 3.5 3. 2.5 2. 1.5 Figure SI-4d. HMBC spectra of compound 4. Intens. x1 6 +MS,.2 #9 511.2 6 475.4 4 2 274.3 31.2 362.3 318.3 385.2 191. 418.8445.3 15 2 25 3 35 4 45 5 m/z Figure SI-4e. (+)ESIMS spectra of compound 4.
S17 of S21 Figure SI-4f. IR spectra of compound 4. Figure SI-4g. CD and UV spectrum of compound 4.
S18 of S21 3 2 1-1 125 1 75 5 25 1 8 6 4 2 DAD1 A, Sig=25,4 Ref=36,1 (H:\2141223.D) 16.273 17.939 19.27 1 2 3 DAD1 A, Sig=25,4 Ref=36,1 (H:\2141228.D) 15.365 17.997 19.656 1 2 3 DAD1 A, Sig=25,4 Ref=36,1 (H:\2141229.D) 15.348 16.33 17.113 18.2 Compd 4 19.647 21.841 D-glc L-glc 22.377 26.112 24.73 25.943 26.985 29.297 34.134 1 2 3 Figure SI-4h. HPLC of compound 4, D-glucose and L-glucose. Table SI-1. Optical rotation of aglycones 1a 4a, moracin O (5), and P (8). Compounds [α] 2 D (c.1 MeOH) [α] 28 D (c.4 MeOH) 1a.328 * 2a.21 * 3a.263 * 4a.95 * Moracin O (5) 5.2 4.2 Moracin P (8) 38.6 15.9 * [α] 2 D can t be calculated due to the non-quantitative deteration.
S19 of S21 Calculated ECD of 1a and MO Analysis Calculated ECD was carried out on R configuration of 1a. Conformation search was done with the MMFF94 using the MOE software package (MOE29.1, Chemical Computing Group, Montreal, QC, Canada). Calculated ECD was performed using TDDFT (Gaussian 9 B.1, Gaussian, Wallingford, CT, 29) at B3LYP/6-31+G(d,p)//B3LYP/6-311+G(d,p) level for the configurations within an energy window of 5 kcal/mol. The conductor-like polarizable continuum model was used with MeOH (ε = 32.613) to take the solvent effects into consideration. The Boltzmann distribution was calculated based on the relative free energy (ΔG), and the final ECD (σ =.2 ev, UV shift = 15 nm) was simulated by using SpecDis (V1.64, University of Wuerzburg, Germany, 215). Table SI-2. The Boltzmann distribution of three conformations (C1 C3) of 1a Conformations of 1a ΔG (Kcal/mol) Boltzmann distribution (%) C1. 39.31 C2.349 21.8 C3.7 38.88 1.5 calcd.1a MO: 86 87.5 exp.1a MO: 86 89 86 9 -.5 1a-C1-1.5 2 25 3 35 4. 1a-C2 1a-C3 MO: 86 89 86 9 MO: 86 87. MO: 86 89 86 9 MO: 86 87 Figure SI-5a. Calculated ECD spectra of 1a-C1 C3.
S2 of S21... MO86: HOMO π of 2-benzylbenzofuran. MO87: LUMO π* of 2-benzylbenzofuran.. MO89: LUMO+2 π* of benzyl moiety. MO9: LUMO+3 π* of benzofuran moiety Figure SI-5b. MO related with two Cotton effects of 1a. HPLC Profile of Cortex Mori Radicis and Their Main Components Identified by HPLC-MS HPLC condition: The mobile solvent was:.1% formic acid (A) and B: MeOH (B). The gradient was: 1% B ( ) 1% B (3 ). The wavelength was 29 nm. The flow rate was 1. ml/. The column was Previl C18 (Grace, 25 4.6mm, 5μ). 7 2.89 23.129 6 3.437 5 4 3 2 1 8.284 8.431 12.36 12.822 14.249 15.271 17.55 17.561 22.381 22.871 24.87 26.831 5 1 15 2 25 3 Figure SI-6. HPLC profile of Cortex Mori Radicis.
S21 of S21 Table SI-3. Main components of Cortex Mori Radicis identified by HPLC-MS R.T. Area Area % M.W. Name Compound N.O. 3.44 1714.9 6.857 46 8.28 832 3.327 568 8.43 1941.7 7.763 ---- 11.7 ---- ---- 344 moracin R 7 12.31 1134.3 4.535 339 12.82 1151.5 4.64 244 oxyresveratrol 6 13.1 ---- ---- 489 2, 3, 4 13.8 ---- ---- 458 1 14.25 724.8 2.898 458 mulberroside C 9 14.6 ---- ---- 32 morin 14 15.27 1229.6 4.916 ---- 17.5 629.5 2.517 326 moracin O 5 17.56 745.6 2.981 326 moracin P 8 18.1 ---- ---- 42 morusin 15 18.3 ---- ---- 562 isomulberrofuran G 11 18.6 ---- ---- 562 mulberrofuran G 12 19.2 ---- ---- 286 norartocarpetin 13 2.81 546.5 2.177 78 Sanggenon D 17 22.38 673.5 2.693 69 cathayanins B 18 22.87 622.6 2.489 776 23.13 532.6 21.21 78 Sanggenon C 16 24.81 54.5 2.161 ---- 26.83 2722.1 1.883 58 The area % was calculated by the area of 29 nm.