S1
Inhibitors of Osteoclast Differentiation from Cephalotaxus koreana
Kee Dong Yoon,† Doc Gyun Jeong,‡ Yun Ha Hwang,‡ Jei Man Ryu,‡ and Jinwoong Kim*,†
College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University, Seoul 151-742, Korea, and Central Research Laboratory, Dong Wha Pharm. Ind. Co. LTD, Anyang-si, Gyunggi-do 430-017, Korea
*
To whom correspondence should be addressed. Phone: +82-2-880-7853. Fax: +82-2887-8509. E-mail:
[email protected]. † Seoul National University. ‡ Central Research Laboratory, Dong Wha Pharm. Ind. Co. Ltd. Supplementary Information S1 Title S2-1 ~ S2-6 Spectroscopic data for compounds 6-24 S3 References for compounds 6-24
S2-1 Apigenin (6)1 yellowish powder; mp: 351-354 oC; [α]24 D : -18.2 (c 0.07, MeOH); UV (MeOH): λmax 268 (2.38), 334 (2.49); IR (KBr) νmax 3363, 2919, 1632, 1510, 1452, 1359, 1256, 1177, 1073 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.90 (d, J = 8.6 Hz, H-2', 6'), 6.91 (d, J = 8.6 Hz, H-3', 5'), 6.76 (s, H-3), 6.45 (d, J = 1.7 Hz, H-8), 6.17 (d, J = 1.7 Hz, H-6); 13C NMR (DMSO, 125 MHz): δ 181.8 (C-4), 164.2 (C-7), 163.8 (C-2), 161.5 (C4'), 161.2 (C-5), 157.4 (C-9), 128.5 (C-2', 6'), 121.2 (C-1'), 116.0 (C-3', 5'), 103.7 (C-10), 102.9 (C-3), 98.9 (C-6), 94.0 (C-8); positive-mode FABMS m/z 271 [M+H]+
→
(7)2 Kaempferol 3-O-α-L-rhamnopyranosyl(1''' 6'')-β-D-glucopyranoside yellowish powder; mp: 178-180 oC; [α]24 D : -2.1 (c 0.14, MeOH); UV (MeOH): λ max (log
ε) 261 (3.04), 346 (2.86) nm; IR (KBr) νmax 3394, 2921, 1653, 1607, 1507, 1453, 1361, 1180, 1069, 838 cm-1; 1H NMR (DMSO, 300 MHz): δ 7.97 (d, J = 8.7 Hz, H- H-2', 6'), 6.87 (d, J = 8.7 Hz, H- H-3', 5'), 6.37 (d, J = 1.8 Hz, H-8), 6.17 (d, J = 1.8 Hz, H-6), 5.29 (d, J = 7.9 Hz, H-1"), 4.39 (brs, H-1'"), 0.99 (d, J = 6.0 Hz, H-6'"); 13C NMR (DMSO, 75 MHz): δ 177.0 (C-4), 164.1 (C-7), 161.0 (C-5), 159.7 (C-4'), 156.4 (C-2), 156.4 (C-9), 133.1 (C-3), 130.6 (C-2',6'), 120.8 (C-1'), 115.8 (C-3',5'), 103.4 (C-10), 101.4 (C-1''), 100.6 (C-1'''), 98.9 (C-6), 93.7 (C-8), 76.3 (C-3''), 75.7 (C-5''), 74.1 (C2''), 71.7 (C-4'''), 70.5 (C-3'''), 70.2 (C-2'''), 69.8 (C-4''), 68.1 (C-5'''), 66.8 (C-6''), 17.5 (C-6'''); positive-ion FABMS m/z 625 [M+Na]+
→
Tamarixetin 3-O-α-L-rhamnopyranosyl(1''' 6'')-β-D-glucopyranoside (8)3 yellowish amorphous powder; mp: 172-174 oC; [α] 24D : 1.4 (c 0.07, MeOH); UV (MeOH): λmax (log ε) 254 (2.32), 351 (2.21) nm; IR (KBr) νmax 3393, 2921, 1653, 1605, 1509, 1358, 1293, 1206, 1066 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.84 (d, J = 1.8 Hz, H-2'), 7.50 (dd, J = 8.7, 1.8 Hz, H-6'), 6.90 (d, J = 8.7 Hz, H-5'), 6.40 (d, J = 1.6 Hz, H8), 6.18 (d, J = 1.6 Hz, H-6), 5.42 (d, J = 6.6 Hz, H-1"), 3.82 (s, 4'-OMe), 0.96 (d, J = 6.2 Hz, H-6'"); 13C NMR (DMSO, 100 MHz): δ 177.2 (C-4), 164.5 (C-7), 161.1 (C-5), 156.4 (C-2), 156.3 (C-9), 149.4 (C-4‘), 146.8 (C-3‘), 133 (C-3), 122.2 (C-6‘), 121.0 (C-1'), 115.2 (C-5‘), 113.3 (C-2'), 103.8 (C-10), 101.2 (C-1''), 100.8 (C-1'''), 98.8 (C-6), 93.8 (C-8), 76.3 (C-3''), 75.8 (C-5''), 74.2 (C-2''), 71.7 (C-4'''), 70.6 (C-3'''), 70.2 (C-2'''), 70.1 (C-4''), 68.2 (C-5'''), 66.7 (C-6''), 55.6 (4'-OMe), 17.6 (C-6''');positive-ion FABMS : m/z 647 [M+Na]+, 625 [M+H]+ Quercetin 3-O-[6"-O-acetyl]-β-D-glucopyranoside (9)2 yellowish powder; mp: 209213 oC; [α]24 D : 6.2 (c 0.07, MeOH); UV (MeOH): λ max (log ε) 257 (2.76), 356 (2.69) nm; IR (KBr) νmax 3392, 1715, 1656, 1607, 1509, 1362, 1300, 1201, 1085 cm-1; 1H NMR
S2-2 (DMSO, 400 MHz): δ 7.53 (dd, J =9.0, 2.0 Hz, H-6'), 7.52 (d, J =2.0 Hz, H-2'), 6.83 (d, J =9.0 Hz, H-5'), 6.40 (d, J =2.0 Hz, H-8), 6.19 (d, J = 2.0 Hz, H-6), 5.37 (d, J =7.3 Hz, H-1"), 1.73 (s,6"-OCOCH3 ); 13C NMR (DMSO, 100 MHz): δ 177.3 (C-4), 169.8 (OCOCH3), 164.2 (C-7), 161.2 (C-5), 156.5 (C-9), 156.3 (C-2), 148.5 (C-4'), 144.8 (C3'), 133.1 (C-3), 121.5 (C-6'), 121.0 (C-1'), 116.1 (C-5'), 115.1 (C-2'), 103.8 (C-10), 100.9 (C-1''), 98.6 (C-6), 93.5 (C-8), 76.2 (C-3''), 73.9 (C-2''), 73.9(C-5''), 69.7 (C-4''), 62.7 (C-6''), 20.1 (OCOCH3); positive-ion FABMS (positive mode): m/z 529 [M+Na]+, 507 [M+H]+ Quercetin 3-O-α-L-rhamnopyranoside (10)2 yellowish powder; mp: 176-179 oC; [α]24 D : -44.3 (c 0.14, MeOH); UV (MeOH): λ max (log ε ) 255 (2.58), 349 (2.43) nm; IR (KBr) νmax 3423, 1653, 1608, 1509, 1455, 1362, 1304, 1202, 997, 816 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.59 (dd, J = 8.0, 1.9 Hz, H-6'), 7.29 (d, J = 1.9 Hz, H-2'), 6.85 (d, J = 8.0 Hz, H-5'), 6.35 (d, J = 1.8 Hz, H-8), 6.17 (d, J = 1.8 Hz, H-6), 0.81 (d, J = 5.84 Hz, H-6"); 13C NMR (DMSO, 100 MHz): δ 177.6 (C-4), 165.1 (C-7), 161.2 (C-5), 157.1 (C-9), 156.5 (C-2), 148.6 (C-4'), 145.2 (C-3'), 134.2 (C-3), 121.1 (C-6'), 120.6 (C1'), 115.5 (C-5'), 115.4 (C-2'), 104.7 (C-10), 101.8 (C-1''), 98.9 (C-6), 93.7 (C-8), 71.2 (C-4''), 70.5 (C-3''), 70.3 (C-2''), 70.1 (C-5''), 17.5 (C-6''); positive-ion FABMS: m/z 449 [M+H]+ Aromadendrin (11)1 pale yellowish powder;: mp: 239-242 oC; [α]24 D 6.98
o
(c 0.07,
MeOH); UV (MeOH): λmax (log ε) 214 (2.26), 291 (2.08) nm; IR (KBr) νmax 3336, 1638, 1515, 1365, 1259, 1169, 1088, 830 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.34 (d, J = 8.4 Hz, H-2', 6'), 6.82 (d, J = 8.4 Hz, H-3', 5'), 5.90 (d, J = 2.0 Hz, H-8), 5.85 (d, J = 2.0 Hz, H-6), 4.95 (d, J = 11.6 Hz, H-2), 4.52 (d, J = 11.6 Hz, H-3); 13C NMR (DMSO, 100 MHz): δ 199.1 (C-4), 170 (C-7), 166.1 (C-5), 165.3 (C-9), 159.9 (C-4'), 131.2 (C-2',6'), 130.1 (C-1'), 116.9 (C-3',5'), 102.5 (C-10), 98.2 (C-6), 97.2 (C-8), 85.7 (C-2), 74.4 (C3); positive-ion FABMS m/z 289 [M+H]+ Apigenin 5-O-β-D-glucopyranoside (12)4 yellowish powder; mp: 237-239 oC; [α]24 D: 19.9o (c 0.09, MeOH); UV (MeOH): λmax (log ε) 260 (2.37), 331 (2.51) nm; IR (KBr) νmax 3430, 1614, 1581, 1462 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.97 (d, J = 8.8 Hz, H-2',6'), 6.90 (d, J = 8.8 Hz, H-3',5'), 6.78 (d, J = 1.6 Hz, H-8), 6.72 (d, J = 1.6 Hz, H-6), 6.65 (s, H-3), 4.69 (d, J = 6.9 Hz, H-1"); 13C NMR (DMSO, 100 MHz): δ 177.0 (C-4), 162.9 (C-7), 161.2 (C-2), 160.8 (C-4'), 158.7 (C-9), 158.4 (C-5), 128.1 (C-2',6'), 121.2 (C-1'), 115.9 (C-3',5'), 108.1 (C-10), 105.6 (C-3), 104.5 (C-6), 104.4 (C-1''), 98.3 (C-8),
S2-3 77.5 (C-5''), 75.6 (C-3''), 73.6 (C-2''), 69.7 (C-4''), 60.8 (C-6''); positive-ion FABMS: m/z 433 [M+H]+, 271 [M-Glc+H]+
→
Apigenin 5-O-α-L-rhamnopyranosyl(1''' 2'')-β-D-glucopyranoside (13)4 yellowish powder; mp: 170-172 oC; [α]24 D : -27.4 (c 0.1, MeOH); UV (MeOH): λ max (log
ε) 213 (2.80), 261 (2.52), 326 (sh, 2.63) nm; IR (KBr) νmax 3361, 1631, 1454, 1604, 1259, 1082 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.84 (d, J = 8.7 Hz, H-2',6'), 6.90 (d, J = 8.7 Hz, H-3',5'), 6.59 (d, J = 1.6 Hz, H-8), 6.53 (d, J = 1.6 Hz, H-6), 6.49 (s, H-3), 5.27 (d, J = 6.8 Hz, H-1"), 5.21 (brs, H-1'"), 1.02 (d, J = 6.1 Hz, H-6'"); 13C NMR (DMSO, 100 MHz): δ 175.6 (C-4), 162.5 (C-7), 160.6 (C-4'), 160.0 (C-2), 158.8 (C-9), 157.6 (C-5), 127.8 (C-2',6'), 121.3 (C-1'), 115.9 (C-3',5'), 107.3 (C-10), 105.9 (C-3), 99.6 (C-1'''), 99.0 (C-6), 97.7 (C-1''), 96.3 (C-8), 77.0 (C-5''), 76.8 (C-3''), 76.4 (C-2''), 72.3 (C-4'''), 70.5 (C-3'''), 70.4 (C-2'''), 69.8 (C-4''), 68.5 (C-5'''), 60.6 (C-6''), 17.9 (C6'''); positive-ion FABMS m/z 601 [M+Na]+
→
Apigenin 7-O-α-L-rhamnopyranosyl(1''' 2'')-β-D-glucopyranoside (14)2 yellowish powder; mp: 245-248 oC; [α]24 D : -7.22 (c 0.05, MeOH); UV (MeOH): λ max (log ε ) 252 (2.70), 333 (2.72) nm; IR (KBr) νmax 3431, 1607, 1508, 1452, 1248, 1179, 1073, 836 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.93 (d, J = 8.8 Hz, H-2', 6'), 6.95 (d, J = 8.8 Hz, H-3',5'), 6.89 (s, H-3), 6.79 (d, J = 2.1 Hz, H-8), 6.36 (d, J = 2.1 Hz, H-6), 5.23 (d, J = 7.1 Hz, H-1"), 5.12 (brs, H-1'"), 1.19 (d, J = 6.2 Hz, H-6'"); 13C NMR (DMSO, 100 MHz): δ 182.0 (C-4), 164.3 (C-2), 162.5 (C-7), 161.4 (C-5), 161.1(C-4'), 157 (C-9), 128.6 (C-2',6'), 121.8 (C-1'), 116.0 (C-3',5'), 105.4 (C-10), 103.2 (C-3), 100.8 (C-1'''), 99.3 (C-6), 97.8 (C-1''), 94.1 (C-8), 77.2 (C-3''), 77.2 (C-5''), 77.0 (C-2''), 71.8 (C4'''), 70.5 (C-2'''), 70.4 (C-3'''), 69.6 (C-4''), 68.3 (C-5'''), 60.4 (C-6''), 18.1 (C-6'''); positive-ion FABMS m/z 579 [M+H]+ Isoscutellarein (15)5 yellowish powder; mp: 252-255 oC; [α]24 D : -2.1 (c 0.07, MeOH); UV (MeOH): λmax (log ε) 220 (2.54), 280 (2.51), 304 (2.51) nm; IR: νmax 3394, 1656, 1580, 1511, 1454, 1366, 1242, 1105, 1007, 832 cm-1; 1H NMR (DMSO, 400 MHz): δ 8.01 (d, J = 8.6 Hz, H-2', 6'), 6.92 (d, J = 8.6 Hz, H-3', 5'), 6.74 (s, H-3), 6.18 (s, H-6); 13 C NMR (DMSO, 100 MHz): δ 182.1 (C-4), 163.7 (C-2), 161.2 (C-4'), 153.4 (C-7), 153.1 (C-5), 145.5 (C-9), 128.6 (C-2',6'), 125.0 (C-8), 121.4 (C-1'), 115.9 (C-3',5'), 103.3 (C-3), 102.3 (C-6), 102.3 (C-10); positive-ion FABMS m/z 287 [M+H]+ Isoscuerllarein 8-O-β-D-glucopyranoside (16)6 yellowish powder; mp: 149-151 oC;
S2-4 [α]24 D : 10.9 (c 0.1, MeOH); UV (MeOH): λmax (log ε) 271 (2.43), 331 (2.39) nm; IR (KBr) νmax 3394, 1654, 1580, 1509, 1362, 1241, 1179, 1068, 1018, 836 cm-1; 1H NMR (DMSO, 400 MHz): δ 8.23 (d, J = 8.8 Hz, H-2'. 6'), 6.93 (d, J = 8.8 Hz, H-3', 5'), 6.80(s, H-3), 6.20 (s, H-6), 4.62 (d, J = 7.7 Hz, H-1"); 13C NMR (DMSO, 100 MHz): δ181.6 (C-4), 163.6 (C-2), 161.2 (C-7), 161.2 (C-4'), 157.3 (C-5), 149.3 (C-9), 129.1 (C-2',6'), 125.7 (C-8), 121.2 (C-1'), 115.9 (C-3',5'), 106.8 (C-1''), 102.7 (C-3), 102.2 (C-10), 99.3 (C-6), 77.2 (C-5''), 76.1 (C-3''), 74.1 (C-2''), 69.0 (C-4''), 60.5 (C-6''); positive-ion FABMS m/z 449 [M+H]+ Luteolin 5-O-β-D-glucopyranoside (17)2 yellowish powder; mp: 268-271 oC; [α]24 D: 8.7 (c 0.07 , MeOH);; UV (MeOH): λmax (log ε) 336 (2.09) nm; IR (KBr) νmax 3366, 1631, 1507, 1449, 1363, 1268, 1173, 1074, 835 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.41 (d, J = 8.2 Hz, H-6'), 7.37 (d, J = 1.7 Hz, H-2'), 6.87 (dd, J = 8.2, 1.7 Hz, H-5'), 6.79 (d, J = 2.0 Hz, H-8), 6.70 (d, J = 2.0 Hz, H-6), 6.54 (s, H-3), 4.70 (d, J = 7.2 Hz, H1"); 13C NMR (DMSO, 100 MHz): δ 176.9 (C-4), 162.7 (C-2), 161.3 (C-7), 158.7 (C-9), 158.3 (C-5), 149.3 (C-4'), 145.7 (C-3'), 121.5 (C-1'), 118.5 (C-6'), 116.0 (C-5'), 113.1 (C-2'), 108.8 (C-10), 105.7 (C-3), 104.6 (C-1''), 104.3 (C-6), 98.2 (C-8), 77.6 (C-5''), 75.6 (C-3''), 73.6 (C-2''), 69.7 (C-4''), 60.8 (C-6''); positive-ion FABMS m/z 449 [M+H]+
→
Luteolin 5-O-α-L-rhamnopyranosyl(1''' 2'')-β-D-glucopyranoside (18)4 yellowish powder; [α]24 D : -8.4 (c 0.07, MeOH); UV (MeOH): λ max (log ε ) 261 (2.23), 336 (2.33) nm; IR (KBr) νmax 3393, 2920, 1634, 1503, 1450, 1358, 1269, 1073, 837 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.32 (d, J = 8.8 Hz, H-6'), 7.31 (d, J = 1.8 Hz, H-2'), 6.85 (dd, J = 8.8, 1.8 Hz, H-5'), 6.56 (d, J = 1.7 Hz, H-8), 6.53 (d, J = 1.7 Hz, H-6), 6.37 (s, H-3), 5.26 (d, J = 6.8 Hz, H-1"), 5.19 (brs, H-1'"), 1.01 (d, J = 6.2 Hz, H-1'"); 13C NMR (DMSO, 100 MHz): δ 175.4 (C-4), 162.2 (C-7), 160.1 (C-2), 158.7 (C-9), 157.6 (C-5), 148.9 (C-4'), 145.6 (C-3'), 121.7 (C-1'), 118.1 (C-6'), 116.0 (C-5'), 112.9 (C-2'), 107.3 (C-10), 105.8 (C-3), 99.5 (C-1'''), 98.9 (C-6), 97.6 (C-1''), 96.0 (C-8), 76.9 (C-5''), 76.7 (C-3''), 76.5 (C-2''), 72.2 (C-4'''), 70.5 (C-2'''), 70.4 (C-3'''), 69.8 (C-4''), 68.6 (C-5'''), 60.6 (C-6''), 17.9 (C-6'''); positive-ion FABMS m/z 617 [M+Na]+ Quercetin 3-O-β-D-glucopyranoside (19)2 yellowish powder; mp: 238-241 oC; [α]24 D: 13.3 (c 0.33, MeOH); UV (MeOH): λmax (log ε) 257 (1.96), 350 (1.89) nm; IR: νmax 3361, 1631, 1454, 1604, 1259, 1082 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.59 (dd, J = 8.9, 2.0 Hz, H-6'), 7.58 (d, J = 2.0 Hz, H-2'), 6.85 (d, J = 8.9 Hz, H-5'), 6.40 (d, J = 1.8
S2-5 Hz, H-8), 6.19 (d, J = 1.8 Hz, H-6), 5.46 (d, J = 7.3 Hz, H-1"); 13C NMR (DMSO, 100 MHz): δ 177.5 (C-4), 164.3 (C-7), 161.3 (C-5), 156.4 (C-9), 156.2 (C-2), 148.5 (C-4'), 144.8 (C-3'), 133.3 (C-3), 121.6 (C-6'), 121.2 (C-1'), 116.2 (C-5'), 115.2 (C-2'), 104.0 (C-10), 100.9 (C-1''), 98.7 (C-6), 93.5 (C-8), 77.6 (C-5''), 76.5 (C-3''), 74.1 (C-2''), 70.0 (C-4''), 61.0 (C-6''); positive-ion FABMS: m/z 487 [M+Na]+
→
(20)2 Quercetin 3-O-α-L-rhamnopyranosyl(1''' 6'')-β-D-glucopyranoside yellowish powder; mp: 202-204 oC; [α]24 D : 3.81 (c 0.05, MeOH); UV (MeOH): λ max (log
ε) 242 (2.02) nm; IR: νmax 3396, 2922, 1651, 1608, 1510, 1363, 1350, 1203, 1071, 814 cm-1; 1H NMR (DMSO, 400 MHz): δ 7.54 (dd, J = 8.1, 2.0 Hz, H-6'), 7.53 (d, J = 2.0 Hz, H-2'), 6.84 (d, J = 8.1 Hz, H-5'), 6.38 (d, J = 1.8 Hz, H-8), 6.19 (d, J = 1.8 Hz, H-6), 5.34 (d, J = 6.6 Hz, H-1"), 5.32 (brs, H-1'"), 0.98 (d, J = 6.0 Hz, H-6'"); 13C NMR (DMSO, 100 MHz): δ 176.4 (C-4), 163.1 (C-7), 160.3 (C-5), 155.7 (C-9), 155.5 (C-2), 147.5 (C-4'), 143.8 (C-3'), 132.7 (C-3), 120.6 (C-1'), 120.2 (C-6'), 115.3 (C-5'), 114.3 (C-2'), 103.1 (C-10), 100.2 (C-1''), 99.8 (C-1'''), 97.7 (C-6), 92.6 (C-8), 75.5 (C-3''), 74.9 (C-5''), 73.1 (C-2''), 70.9 (C-4'''), 69.6 (C-3'''), 69.4 (C-2'''), 69.0 (C-4''), 67.3 (C-5'''), 66.0 (C-6''), 16.8 (C-6'''); positive-ion FABMS m/z 633 [M+Na]+, 611 [M+H]+ (+)-Catechin (21)7 brownish powder; mp: 170-173 oC; [α]24 D : 3.3 (c 0.04, MeOH); UV (MeOH): λmax (log ε) 215 (3.35), 279 (2.58) nm; IR (KBr) νmax 3393, 1615, 1520, 1465, 1369, 1282, 1143, 1103, 1038, 819 cm-1; 1H NMR (DMSO, 400 MHz): δ 6.83 (d, J = 1.8 Hz, H-2'), 6.75 (d, J = 8.0 Hz, H-5'), 6.70 (dd, J = 8.0, 1.8 Hz, H-6'), 5.92 (d, J = 2.4 Hz, H-8), 5.85 (d, J = 2.4 Hz, H-6), 4.57 (d, J = 7.3 Hz, H-2), 3.97 (ddd, J = 8.0, 7.3, 5.5 Hz, H-3), 2.84 (dd, J = 16.1, 5.5 Hz, H-4a), 2.50 (dd, J = 16.1, 8.0 Hz, H-4b); 13C NMR (DMSO, 100 MHz): δ 156.4 (C-5), 156.1 (C-9), 155.3 (C-7), 144.8 (C-3'), 144.8 (C-4'), 130.6 (C-1'), 118.3 (C-6'), 115.0 (C-5'), 114.5 (C-2'), 99.0 (C-10), 95.1 (C-6), 93.8 (C-8), 81.0 (C-2), 66.3 (C-3), 27.8 (C-4); positive-ion FABMS m/z 291 [M+H]+ Catechin 7-O-β-D-glucopyranoside (22)2 brownish powder; mp: 216-218 oC; [α]24 D: 3.9 (c 0.03, MeOH); UV (MeOH): λmax (log ε) 281 (2.17) nm; IR: νmax 3368 (OH), 1614, 1459, 1367cm-1; 1H NMR (DMSO, 400 MHz): δ 6.85 (d, J = 1.9 Hz, H-2'), 6.75 (d, J = 8.1 Hz, H-5'), 6.70 (dd, J = 8.1, 1.9 Hz, H-6'), 6.19 (d, J = 2.3 Hz, H-8), 6.15 (d, J = 2.3 Hz, H-6), 4.81 (d, J = 7.4 Hz, H-1"), 4.57 (d, J = 7.5 Hz, H-2), 3.98 (ddd, J = 8.2, 7.5, 5.4 Hz, H-3), 2.85 (dd, J = 16.1, 5.4 Hz, H-4a), 2.53 (dd, J = 16.1, 8.2 Hz, H-4b); 13C NMR (DMSO, 100 MHz): δ 159.4 (C-7), 158.3 (C-5), 157.4 (C-9), 147.0 (C-3'), 147.0 (C-4'), 132.8 (C-1'), 120.8 (C-6'), 116.9 (C-5'), 116.0 (C-2'), 104.3 (C-10), 102.9 (C-1''),
S2-6 98.1 (C-8), 97.6 (C-6), 83.7 (C-2), 78.7 (C-3''), 78.7 (C-5''), 75.5 (C-2''), 72.0 (C-4''), 69.3 (C-3), 63.2 (C-6''), 29.3 (C-4); positive-ion FABMS m/z 453 [M+H]+ (-)-Epicatechin (23)7 brownish powder; mp: 235-237 oC; [α]24 D : -3.8 (c 0.07, MeOH); UV (MeOH): λmax (log ε) 280 (1.79) nm; IR (KBr) νmax 3363, 1615, 1518, 1461, 1365, 1281, 1146, 1093, 826 cm-1; 1H NMR (DMSO, 400 MHz): δ 6.96 (d, J = 1.8 Hz, H-2'), 6.79 (dd, J = 8.3, 1.8 Hz, H-6'), 6.75 (d, J = 8.3 Hz, H-5'), 5.93 (d, J = 2.2 Hz, H-8), 5.90 (d, J = 2.2 Hz, H-6), 4.81 (brs, H-2), 3.97 (m, H-3), 2.86 (dd, J = 16.7, 4.6 Hz, H-4a), 2.72 (dd, J = 16.1, 2.8 Hz, H-4b); 13C NMR (DMSO, 100 MHz): δ 156.4 (C-5), 156.2 (C-7), 155.7 (C-9), 144.5 (C-4'), 144.4 (C-3'), 130.6 (C-1'), 117.9 (C-6'), 114.9 (C-2'), 114.9 (C-5'), 98.4 (C-10), 95.1 (C-6), 94.0 (C-8), 78.0 (C-2), 64.9 (C-3), 28.1 (C-4); positive-ion FABMS m/z 291 [M+H]+ Puerarin (24)8 pale yellowish powder; mp: 190-193 oC; [α]24 D : 12.1 (c 0.07, MeOH); UV (MeOH): λmax (log ε) 249 (2.67), 303 (2.20) nm; IR (KBr) νmax 3364, 1628, 1514, 1447, 1398, 1263 cm-1; 1H NMR (DMSO, 400 MHz): δ 8.16 (s, H-2), 8.04 (d, J = 9.1 Hz, H-5), 7.36 (d, J = 8.7 Hz, H-2', 6'), 6.97 (d, J = 9.1 Hz, H-6), 6.83 (d, J = 8.7 Hz, H3', 5'), 5.09 (d, J = 9.6 Hz, H-1"); 13C NMR (DMSO, 100 MHz): δ 178.3 (C-4), 163.3 (C-7), 158.7 (C-4'), 158.6 (C-9), 154.4 (C-2), 131.4 (C-2', 6'), 128.1 (C-5), 125.6 (C1'), 124.2 (C-3), 118.4 (C-10), 116.3 (C-6), 116.2 (C-3', 5'), 113.1 (C-8), 82.8 (C-5''), 80.0 (C-3''), 75.7 (C-1''), 73.0 (C-2''), 71.7 (C-4''), 62.7 (C-6''); positive-ion FABMS m/z 417 [M+H]+
S3
References (1) Agrawal, P. K.; Bansal, M. C. Carbon-13 NMR of Flavonoids: Flavonoids; Agrawal, P. K. Ed.; Elservier: Amsterdam, 1989; pp 95-173. (2) Agrawal, P. K.; Bansal, M. C. Carbon-13 NMR of Flavonoids: Flavonoid glycoside; Agrawal, P. K. Ed.;. Elservier: Amsterdam, 1989; pp 283-355. (3) Sastry, C. V. R.; Reddy, C.; Rukmini, C.; Row, L. R. Indian J. Chem. 1967, 5, 613615. (4) Ling, S. K.; Takashima, T.; Tanaka, T.; Fujioka, T.; Mihashi, K.; Kouno, I. Fitoterapia 2004, 75, 785-788. (5) Markham, K. R.; Porter, L. J. Phytochemistry, 1975, 14, 1093-1097. (6) Kamiya, K.; Saiki, Y.; Hama, T.; Fujimoto, Y.; Endang, H.; Umar, M.; Satake, T. Phytochemistry 2001, 57, 297-301. (7) Agrawal, P. K.; Bansal, M. C. Carbon-13 NMR of Flavonoids: Flavanoids; Agrawal, P. K. Ed.; Elservier: Amsterdam, 1989; pp 432-496. (8) Lee, H. J.; Oh, M. A.; Choi, Y. H.; Lee, K. M. Yakhak Hoeji 2001, 45, 500-505.
