Facile Synthesis of Flavonoid 7-O-Glycosides

Facile Synthesis of Flavonoid 7-O-Glycosides Ming Li, a Xiuwen Han, a Biao Yu b * a State Key Laboratory of Catalyst, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China b State Key Laboratory of Bio-organic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China. Fax: (86)21-64166128, email: byu@mail.sioc.ac.cn Supporting Information Experimental 1. General remarks. Solvents were distilled from the appropriate drying agents before use. Unless stated otherwise, all reactions were monitored by TLC on silica gel HF 254 (0.5 mm, Qingdao, China). Spots were detected under UV light or by charging with 10% H 2 SO 4 in MeOH. Flash column chromatography was carried out on silica gel H (400 mesh, Qingdao, China). Optical rotations were measured on a Perkin-Elmer 241 MC polarimeter. 1 H NMR spectra were recorded on a Bruker AM-300 spectrometer using tetramethylsilane as an internal reference. Mass spectra were recorded on a HP5989A mass spectrometer. Elemental analyses were performed on a Perkin-Elmer Model 2400 instrument. 2. General procedure for acylation of flavonoids 2.1 4, 7, 8-Trihexanoxy isoflavone (4) Method A: To a solution of 4, 7, 8-trihydroxy isoflavone (1.08 g, 4.00 mmol) in pyridine (10 ml) and CHCl 3 (5 ml) was added DMAP (155 mg, 1.27 mmol, 0.3 equiv.) and hexanoyl chloride (3.2 ml, 22.89 mmol, 5.7 equiv.) at -20 o Cintwo portions. After stirring for 9 h, the mixture was diluted with CH 2 Cl 2, and washed with 1 M aq. HCl and saturated brine subsequently. The organic phase was dried over anhydrous Na 2 SO 4 and then concentrated. The residue was applied to a silica gel column chromatography (petroleum ether-etoac, 9:1) to afford 4 as a white solid (1.97 g, 87%): 1 H NMR (300 MHz, CDCl 3 ): 8.19 (d, 1 H, J = 9.0 Hz), 7.96 (s, 1 H), 7.55 1

(d, 2 H, J = 8.4 Hz), 7.25 (d, 1 H, J = 8.7 Hz), 7.16 (d, 2 H, J = 8.4 Hz), 2.67 (t, 2 H, J = 7.8 Hz), 2.62-2.55 (m, 4 H), 1.81-1.75 (m, 6 H), 1.44-1.38 (m, 12 H), 0.97-0.91 (m, 9 H). 13 CNMR(75MHz,CDCl 3 ): 175.01, 172.13, 170.32, 170.02, 152.55, 150.97, 149.53, 146.78, 130.00, 128.68, 124.99, 123.78, 123.14, 121.71, 120.19, 34.35, 33.97, 33.64, 31.18, 24.58, 24.50, 22.24, 13.83. EI-MS (m/z): 466, 368, 270, 99, 71. Anal. Calcd for C 33 H 40 O 8 : C, 70.19%, H, 7.14%; Found: C, 70.10%, H, 7.24%. 2.2 Chrysin benzoate (6c) Method B: To a solution of Chrisin (1.00 g, 3.93 mmol) in anhydrous pyridine (7 ml), benzoyl chloride (1 ml) was added at 0 o C. After stirring at 60 o C for 7 h, TLC showed that the reaction was completed. The mixture was quenched with methanol and then concentrated and diluted with CH 2 Cl 2. The organic phase, after washing with 1N aq. HCl and saturated brine sequentially, was dried over anhydrous Na 2 SO 4 and concentrated. The residue was crystallized in petroleum ether-etoac (1:2) to produce 6c as a white solid (1.23 g, 95%). 1 H NMR (300 MHz, CHCl 3 ): 8.29 (d, 2 H, J =7.4 Hz), 8.23 (d, 2 H, J = 7.4 Hz), 7.89 (dd, 2 H, J = 2.2, 7.0 Hz), 7.70-7.54 (m, 10 H), 7.15 (d, 1 H, J = 2.2 Hz), 6.66 (s, 1 H). EI-MS (m/z): 463 (M + H + ), 105, 77. Anal. Calcd for C 29 H 18 O 6 0.25H 2 O: C, 74.59%, H, 3.99%; Found: C, 74.85%, H, 3.72%. 2.3 Chrysin hexanoate (6a) Method B was used to provide 6c (87%) as a white powder: 1 H NMR (300 MHz, CDCl 3 ): 7.87-7.85 (m, 2 H), 7.53-7.49 (m, 3 H), 7.35 (d, 1 H, J = 2.2 Hz), 6.83 (d, 1 H, J = 2.0 Hz), 6.66 (s, 1 H), 2.74 (t, 2 H, J = 7.7 Hz), 2.60 (t, 2 H, J = 7.5 Hz), 1.87-1.73 (m, 4 H), 1.46-1.38 (m, 8 H), 0.96-0.92 (m, 6 H). EI-MS (m/z): 352, 254. Anal. Calcd for C 27 H 30 O 6 : C, 71.98%, H, 6.71%; Found: C 72.00%, H, 6.89%. 2.4 Chrysin acetate (6b) Ref. 11. 2.5 Daidzein hexanoate (8a) 2

Method A was used to prepare 8c (77%) as a white solid from daidzein. 1 H NMR (300 MHz, CDCl 3 ): 8.33 (d, 1 H, J = 8.8 Hz), 8.02 (s, 1 H), 7.59 (d, 2 H, J = 8.8 Hz), 7.31 (d, 1 H, J = 2.2 Hz), 7.20-7.16 (m, 3 H), 2.62 (t, 2 H, J = 7.4 Hz), 2.58 (t, 2 H, J =7.7 Hz), 1.80-1.76 (m, 2 H), 1.45-1.39 (m, 8 H), 0.98-0.92 (m, 6 H). EI-MS (m/z): 451 [M +H + ], 352, 353, 254, 99, 77. Anal. Calcd for C 27 H 30 O 6 : C, 71.98%, H, 6.71%; Found: C 72.17%, H, 6.97%. 2.6 Daidzein acetate (8b) Ref. 10. 2.7 Quercetin acetate (10a) Ref.: Picq, M.; Prigent, A. F.; Némoz, G.; André, A. C.; Pacheco, H. J. Med. Chem. 1982, 25, 1192. 2.8 Quercetin benzoate (10b) Method B was used to prepare 10b (56%) as a white solid: 1 H NMR (300 MHz, DMSO-d 6 ): 8.14-7.20 (m, 30 H). EI-MS (m/z): 823 (M + H + ), 105, 77. Anal. Calcd for C 50 H 30 O 12 : C, 72.99%, H, 3.68%; Found: C 73.16%, H, 3.57%. 2.9 7,8-Dihexanoxy chromone (14). Method A was used to prepare 14 (85%) as an orange syrup: 1 H NMR (300 MHz, CDCl 3 ): 8.10 (d, 1 H, J = 9.0 Hz), 7.80 (d, 1 H, J = 6.3 Hz), 7.23 (d, 1 H, J =9.0Hz), 6.34 (d, 1 H, J = 6.3 Hz), 2.65 (t, 2 H, J = 7.2 Hz), 2.58 (t, 2 H, J = 7.5 Hz), 1.80-1.76 (m, 4 H), 1.43-1.37 (m, 8 H), 0.96-0.91 (m, 6 H); EI-MS (m/z): 375, 276, 178, 99, 71, 43. Anal. Calcd for C 21 H 26 O 6 : C, 67.36%, H, 7.00%; Found: C 66.94%, H, 7.30%. 3. Preparation of 7,8-dihexanoxyisoflavone 4 -O- 2, 3, 5 -tri-o-benzoyl- -Darabinofuranoside (12) 3.1 7, 8-Dihexanoxy-4 -hydoxy isoflavone (28) To a solution of 4, 7, 8-trihydroxy isoflavone (806 mg, 2.98 mmol) in pyridine (8 ml) and CHCl 3 (4 ml) at -78 o C was added DMAP (93 mg, 0.76 mmol, 0.26 equiv.) and 3

hexanoyl chloride (0.9 ml, 6.44 mmol, 2.16 equiv.). The mixture was warmed to 0 o C in a period of 10 h, and then diluted with CH 2 Cl 2. The organic phase, after washing with 1 M aq. HCl and saturated brine subsequently, was dried over anhydrous Na 2 SO 4 and then concentrated. The residue was applied to a silica gel column chromatography (petroleum ether-etoac, 3:1) to afford 28 as a white solid (1.06 g, 76%): 1 H NMR (300 MHz, DMSO-d 6 ): 9.64 (s, 1 H), 8.46 (s, 1 H), 8.08 (d, 1 H, J = 8.7 Hz), 7.46 (d, 1 H, J = 8.7 Hz), 7.42 (d, 2 H, J = 8.4 Hz), 6.85 (d, 2 H, J = 8.4 Hz), 2.72 (t, 2 H, J =7.5 Hz), 2.66 (t, 2 H, J = 7.5 Hz), 1.72-1.63 (m, 4 H), 1.41-1.33 (m, 8 H), 0.93-0.89 (m, 6 H). 13 CNMR(75MHz,CDCl 3 ): 175.92, 170.60, 170.28, 156.57, 152.43, 149.54, 146.66, 131.41, 130.15, 125.52, 123.82, 123.03, 122.61, 120.10, 115.70, 33.97, 33.66, 31.17, 24.58, 24.49, 22.23, 13.83. ESI-MS (m/z): 489.2 [M + Na + ], 467.2 [M + H + ]. Anal. Calcd for C 27 H 30 O 7 0.5H 2 O: C, 68.19%, H, 6.57%; Found: C 68.56%, H, 6.54%. 3.2 7,8-Dihexanoxyisoflavone 4 -O- 2, 3, 6 -tri-o-benzoyl- -D-arabinofuranoside (12) See the general procedure for the glycosylation (Section 6). The coupling of 28 with 18 produced 12 in 88% yield as a foam: [ ] 20 D = 34.1 (c 1.04, CHCl 3 ); 1 H NMR (300 MHz, CDCl 3 ): 8.21 (d, 1 H, J = 8. 8 Hz), 8.16-8.13 (m, 2 H), 8.08-8.04 (m, 4 H), 7.96 (s, 1 H), 7.66-7.22 (m, 14 H), 6.04 (brs, 1 H), 5.83 (brs, 1 H), 5.72 (d, 1 H, J =3.3 Hz), 4.85-4.73 (m, 3 H), 2.68 (t, 2 H, J = 7.4 Hz), 2.61 (t, 2 H, J = 7.7 Hz), 1.85-1.76 (m, 4 H), 1.48-1.40 (m, 8 H), 0.99-0.87 (m, 6 H). 13 CNMR(75MHz,CDCl 3 ): 175.22, 170.35, 170.05, 166.16, 165.72, 165.33, 156.22, 152.19, 149.34, 146.69, 133.61, 133.05, 130.24, 129.95, 129.77, 129.10, 128.88, 128.56, 128.31, 125.46, 125.20, 123.81, 123.15, 120.09, 116.90, 104.06, 82.19, 82.10, 77.68, 63.64, 33.99, 33.67, 31.18, 24.61, 24.52, 22.26, 13.86. EI-MS (m/z): 445, 368, 270, 105, 77, 71, 43. Anal. Calcd for C 53 H 50 O 14 : C, 69.88%, H, 5.53%; Found: C, 69.84%, H, 5.48%. 4. General procedure for the regioselective deacylation To a solution of the flavonoid ester (1 mmol) in anhydrous NMP at 0 o Cwasadded imidazole (0.35 equiv.) followed by PhSH (1.2 molar equiv.). After stirring at 25 o C until the disappearance of the starting material as monitored by TLC, the mixture was 4

diluted with CH 2 Cl 2, and then washed with 1 N aq. HCl and saturated brine subsequently. The organic phase was dried over anhydrous Na 2 SO 4 and concentrated. The residue was applied to a silica gel column chromatography to afford the product. 4.1 4, 8-Dihexanoxy-7-hydroxy isoflavone (5) Compound 5 was obtained as a white solid: 1 H NMR (300 MHz, DMSO-d 6 ): 11.11 (s,1h),8.42(s,1h),7.87(d,1h,j = 9.3 Hz), 7.58 (d, 2 H, J = 8.7 Hz), 7.15-7.08 (m,3h),2.67(t,2h,j = 7.4 Hz), 2.57 (t, 2 H, J = 7.4 Hz), 1.71-1.63 (m, 4 H), 1.41-1.30 (m, 8 H), 0.91-0.87 (m, 6 H); 13 C NMR (75 MHz, DMSO-d 6 ): 174.32, 171.97, 170.92, 154.56, 153.74, 150.38, 130.26, 129.36, 125.59, 123.52, 123.22, 121.70, 117.09, 115.37, 33.62, 33.16, 30.78, 30.68, 24.31, 24.19, 21.94, 13.96; ESI-MS (m/z): 467.2 [M + H + ]. Anal. Calcd for C 27 H 30 O 7 : C, 69.51%, H, 6.48%; Found: C 69.49%, H, 6.57%. Phenylthiohexanoate was isolated as a liquid: 1 H NMR (300 MHz, CDCl 3 ): 7.42 (m, 5 H), 2.66 (t, 2 H, J = 7.2 Hz), 1.57 (m, 2 H), 1.36 (m, 4 H), 0.91 (t, 3 H, J =6.9Hz); EI-MS (m/z): 208 (M + ), 109, 99, 71, 43. Anal. Calcd for C 12 H 16 OS: C, 69.11%, H, 7.74%, S 15.39%; Found: C 68.90%, H, 7.70%, S 15.06%. 4.2 5-O-Hexanoyl chrysin (7a) Compound 7a was obtained as a white powder: 1 H NMR (300 MHz, DMSO-d 6 ): 11.12 (s, 1 H), 8.03 (m, 2 H), 7.58 (m, 3 H), 6.94 (d, 1 H, J = 2.2 Hz), 6.77 (s, 1 H), 6.55 (d, 1 H, J = 2.5 Hz), 2.62 (t, 2 H, J = 7.4 Hz), 1.67 (m, 2 H), 1.36 (m, 4 H), 0.94 (t, 3 H, J =7.0Hz).ESI-MS(m/z): 353.1 [M + H + ]. Anal. Calcd for C 21 H 20 O 5 :C, 71.58%, H, 5.72%; Found: C 71.49%, H, 5.83%. 4.3 5-O-Acetyl chrysin (7b) Ref. 11. 4.4 5-O-Benzoyl chrysin (7c) 5

Compound 7c was obtained as a white powder: 1 H NMR (300 MHz, DMSO-d 6 ): 11.86 (s, 1 H), 8.13-8.03 (m, 4 H), 7.75-7.55 (m, 6 H), 7.01 (m, 1 H), 6.74 (m, 1 H), 6.70 (s, 1 H). ESI-MS: 381.1 [M + Na + ], 359.1 [M + H + ]. Anal. Calcd for C 22 H 14 O 5 :C, 73.74%, H, 3.94%; Found: C 73.30%, H, 3.97%. 4.5 4 -O-Hexanoyl daidzein (9a) Compound 9a was obtained as a white solid: 1 H NMR (300 MHz, DMSO-d 6 ): 10.84 (s,1h),8.42(s,1h),7.99(d,1h,j = 8.8 Hz), 7.61 (d, 2 H, J = 8.5 Hz), 7.17 (d, 2 H, J = 8.5 Hz), 6.98-6.89 (m, 2 H), 2.59 (t, 2 H, J = 7.4 Hz), 1.68-1.61 (m, 2 H), 1.37-1.32 (m,4h),0.90(t,3h,j =6.6Hz);ESI-MS(m/z): 353.2 [M + H + ]. Anal. Calcd for C 21 H 20 O 5 0.25H 2 O C, 70.67%, H, 5.79%; Found: C 70.91%, H, 5.78%. 4.6 4 -O-acetyl daidzein (9b) Ref.: 10. 4.7 3, 3, 4, 5-Tetra-O-acetyl quercetin (11a) Ref.: Natoli, M.; Nicolosi, G.; Piattelli, M. J. Org. Chem. 1992, 57, 5776. 4.8 3, 3, 4, 5-Tetra-O-benzoyl quercetin (11b) Compound 11b was obtained as a white solid: 1 H NMR (300 MHz, DMSO-d 6 ): 11.47 (s,1h),8.19(d,1h,j = 1.8 Hz), 8.11 (d, 4 H, J = 7.2 Hz), 8.02-7.92 (m, 5 H), 7.74-7.44 (m, 13 H), 7.08 (d, 1 H, J = 2.1 Hz), 6.86 (d, 1 H, J =2.4Hz). 13 CNMR(75MHz, DMSO-d 6 ): 168.99, 164.39, 163.48, 163.38, 163.22, 163.06, 157.82, 152.84, 150.46, 144.54, 142.54, 134.51, 133.86, 133.16, 130.15, 130.08, 129.74, 129.23, 129.05, 128.88, 127.95, 127.82, 126.98, 124.61, 123.86, 109.89, 109.44, 101.53. ESI-MS (m/z): 719.2 [M + H + ]. Anal. Calcd for C 43 H 26 O 11 0.5H 2 O: C, 70.97%, H, 3.74%; Found: C 70.95%, H, 3.43%. 4.9 8-Hexanoxy-7-hydroxy isoflavone 4 -O-2, 3, 5 -tri-o-benzoyl- -Darabinofuranoside (13) 6

Compound 13 was obtained as a semi-solid: 1 H NMR (300MHz, DMSO-d 6 ): 11.08 (s, 1 H), 8.37 (s, 1 H), 8.10-7.96 (m, 6 H), 7.88 (d, 1 H, J = 9.0 Hz), 7.74-7.51 (m, 9 H),7.37(d,2H,J = 7.8 Hz), 7.24 (d, 1 H, J = 8.1 Hz), 7.10 (d, 1 H, J = 9.3 Hz), 6.32 (brs, 1 H), 5.68 (m, 2 H), 4.71 (m, 3 H), 2.66 (t, 2 H, J = 7.2 Hz), 1.69-1.62 (m, 2 H), 1.36-1.29 (m, 4 H), 0.87 (t, 3 H, J =7.2Hz).ESI-MS(m/z): 277.1 [M + H + ]. Anal. Calcd for: C 47 H 40 O 13 0.5H 2 O: C, 68.69%, H, 5.03%; Found: C 68.61%, H, 5.34%. 4.10 8-Hexanoxy-7-hydroxy chromone (15) Compound 15 was obtained as a yellow solid: 1 H NMR (300 MHz, DMSO-d 6 ): 11.06 (s,1h),8.17(d,1h,j = 6.3 Hz), 7.77 (d, 2 H, J = 9.3 Hz), 7.07 (d, 1 H, J =9.0Hz), 6.26 (d, 1 H, J = 6.3 Hz), 2.68 (t, 2 H, J = 7.4 Hz), 1.71-1.66 (m, 2 H), 1.42-1.33 (m, 4 H),0.90(m,3H).ESI-MS(m/z): 277.1 [M + H + ]. Anal. Calcd for C 15 H 16 O 5 :C, 65.21%, H, 5.84%; Found: C 65.23%, H, 5.82%. 5.1 2,3,4,6-Tetra-O-benzoyl-D-glucopyranosyl (N-phenyl)trifluoroacetimidate (16) Ref. 20b 5.2 2,3,4-Tri-O-acetyl-L-rhamnopyranosyl (N-phenyl)trifluoroacetimidate (17) Ref. 20b 5.3 2,3,5-Tri-O-benzoyl-D-arabinofuranosyl (N-phenyl)trifluoroacetimidate (18) Compound 18 (939 mg, 92%) was prepared as a white foam from 2,3,5-tri-O-benzoyl- D-arabinofuranoside using the procedure described in ref. 20b. Several mg of the pure -isomer was obtained for characterization: [ ] 20 D = -23.8 (c 1.40, CHCl 3 ). 1 HNMR (300 MHz, CDCl 3 ): 8.12-8.00 (m, 6 H), 7.65-7.26 (m, 11 H), 7.09 (t, 1 H, J =7.5 Hz), 6.88 (d, 2 H, J = 7.5 Hz), 6.65 (brs, 1 H), 5.81 (brs, 1 H), 5.68 (d, 1 H, J =2.7 Hz), 4.85-4.69 (m, 3 H). Anal. Calcd for C 34 H 26 NO 8 F 3 :C,64.46%,H,4.14%,N, 2.21%; Found: C 64.29%, H, 4.10%, N, 2.40%. 6 General procedure for glycosylation 7

To a suspension of the flavonoid acceptor (0.1 mmol), glycosyl trifluoroacetimidate donor (0.25 mmol), and 4Å molecular sieves in CH 2 Cl 2 (5mL)wasaddedasolutionof BF 3 Et 2 OinCH 2 Cl 2 (0.1 M 0.3 ml). After stirring at room temperature overnight, the reaction was quenched with Et 3 N. The mixture was then filtrated and concentrated. The residue was purified on a silica gel column chromatography to give the glycoside. 6.1.1 4, 8-Dihexanoxyisoflavone 7-O-2,3,4,6 -tetra-o-benzoyl- -D-glucopyranoside (19) Compound 19 was obtained as a white foam: [ ] 20 D =23.3 (c 1.06, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.06-7.84 (m, 10 H), 7.61-7.29 (m, 15 H), 7.14 (d, 2 H, J =8.4 Hz), 6.02 (t, 1 H, J = 9.6 Hz), 5.87 (dd, 1 H, J = 7.8, 9.3 Hz), 5.70 (t, 1 H, J =9.6Hz), 5.52 (d, 1 H, J =7.2 Hz), 4.73 (dd, 1 H, J = 2.4, 9.9 Hz), 4.53 (dd, 1 H, J = 6.9, 12.0 Hz), 4.41 (m, 1 H), 2.59-2.43 (m, 4 H), 1.79-1.55 (m, 4 H), 1.42-1.33 (m, 8 H), 0.96-0.89 (m, 6 H). 13 CNMR(75MHz,CDCl 3 ): 175.00, 172.24, 170.49, 165.91, 165.60, 165.21, 165.00, 152.54, 152.43, 150.82, 149.61, 133.66, 133.55, 133.47, 133.40, 130.00, 129.88, 129.82, 129.74, 129.63, 128.77, 128.53, 128.37, 124.63, 124.11, 121.71, 120.60, 113.21, 99.08, 73.01, 72.54, 71.31, 69.35, 62.96, 34.34, 33.20, 31.21, 31.15, 24.56, 24.20, 22.27, 13.94. EI-MS (m/z): 466, 368, 270, 105, 99, 77, 71, 43. Anal. Calcd for: C 61 H 56 O 16 C, 70.11%, H, 5.40%; Found: C 69.74%, H, 5.52%. 6.1.2 4, 7-Dihexanoxyisoflavone 8-O-2,3,4,6 -tetra-o-benzoyl- -D-glucopyranoside (19a) Compound 19a was obtained as a white foam: [ ] 20 D =1.2(c 1.15, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.07-7.87 (m, 9 H), 7.46-7.22 (m, 15 H), 7.14 (d, 2 H, J =8.4 Hz), 7.10 (d, 1 H, J = 9.0 Hz), 6.02 (t, 1 H, J = 9.6 Hz), 5.92-5.82 (m, 2 H), 5.37 (d, 1 H, J = 7.5 Hz), 4.53 (m, 2 H), 4.11 (m, 1 H), 2.61-2.46 (m, 4 H), 1.81-1.76 (m, 2 H), 1.63-1.57 (m, 2 H), 1.44-1.40 (m, 4 H), 1.26-1.19 (m, 4 H), 0.95 (t, 3 H, J =6.6Hz), 0.89 (t, 3 H, J =6.9Hz). 13 C NMR (75 MHz, CDCl 3 ): 174.97, 172.25, 171.07, 165.91, 165.78, 165.09, 164.96, 152.48, 150.84, 150.30, 147.98, 136.62, 133.55, 133.41, 133.29, 129.80, 129.62, 129.34, 128.59, 128.45, 124.49, 123.26, 122.79, 8

121.65, 120.78, 113.21, 102.76, 72.68, 72.20, 69.02, 34.38, 33.91, 31.26, 31.11, 24.61, 24.28, 22.30, 13.91. Anal. Calcd for C 61 H 56 O 16 : C, 70.11%, H, 5.40%; Found: C, 69.83%, H, 5.69%. 6.2 4, 8-Dihexanoxyisoflavone 7-O-2,3,4 -tri-o-acetyl- -L-rhamnopyranoside (20) Compound 20 was obtained as a white foam: [ ] 20 D =-64.1(c 1.18, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.14 (d, 1 H, J = 8.1 Hz), 7.95 (s, 1 H), 7.55 (d, 2 H, J =8.4Hz), 7.30 (d, 1 H, J = 8.1 Hz), 7.15 (d, 2 H, J = 8.4 Hz), 5.60 (brs, 1 H), 5.44 (m, 1 H), 5.37 (dd, 1 H, J =3.0,10.2Hz),5.17(t,1H,J = 9.9 Hz), 2.81 (t, 2 H, J = 7.4 Hz), 2.57 (t, 2 H, J = 7.5 Hz), 2.20 (s, 3 H), 2.07 (s, 3 H), 2.02 (s, 3 H), 1.84-1.77 (m, 4 H), 1.41 (m, 8 H), 1.21(t,3H,J = 6.0 Hz), 0.93 (m, 3 H). 13 C NMR (75 MHz, CDCl 3 ): 175.10, 172.21, 171.14, 169.85, 152.57, 151.35, 150.85, 149.68, 133.66, 130.03, 128.86, 128.21, 124.70, 124.28, 121.71, 120.32, 113.21, 95.91, 70.20, 69.11. 68.56, 68.06, 34.34, 33.41, 31.15, 24.58, 24.41, 22.27, 20.72, 20.60, 17.35, 13.94. EI-MS (m/z): 467, 368, 273, 153, 111, 99, 71, 43. Anal. Calcd for C 39 H 46 O 14 :C,63.41%,H,6.28%; Found: C, 63.15%, H, 6.48%. 6.3.1 4, 8-Dihexanoxyisoflavone 7-O-2,3,5 -tri-o-benzoyl- -D-arabinofuranoside (21) Compound 21 was obtained as a white foam: [ ] 20 D = 34.5 (c 0.91, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.19 (d, 1 H, J = 8.1 Hz), 8.12 (m, 2 H), 8.06-8.03 (m, 4 H), 7.95 (s, 1 H), 7.65-7.30 (m, 12 H), 7.17 (d, 2 H, J = 8.7 Hz), 6.09 (brs, 1 H), 5.82 (brs, 1 H), 5.72 (d, 1 H, J = 3.3 Hz), 4.85-4.73 (m, 3 H), 2.64-2.57 (m, 4 H), 1.81-1.76 (m, 4 H), 1.47-1.42 (m, 4 H), 1.36-1.30 (m, 4 H), 1.00-0.89 (m, 6 H); 13 CNMR(75MHz, CDCl 3 ): 175.10, 172.14, 170.64, 166.06, 165.65, 165.21, 152.46, 152.05, 150.88, 149.73, 133.72, 133.14, 129.95, 129.72, 129.57, 128.90, 128.59, 128.34, 124.72, 124.22, 121.67, 120.47, 114.31, 104.90, 82.68, 81.99, 63.30, 34.35, 33.64, 31.21, 31.12, 24.61, 24.20, 22.26, 22.15, 13.83. EI-MS (m/z): 466, 445, 368, 270, 105, 99, 77, 71, 43. Anal. Calcd for C 53 H 50 O 14 : C, 69.88%, H, 5.53%; Found: C, 69.60%, H, 5.53%. 9

6.3.2 4, 7-Dihexanoxyisoflavone 8-O-2,3,5 -tri-o-benzoyl- -D-arabinofuranoside (21a) Compound 21a was obtained as a white foam: [ ] 20 D = 18.3 (c 0.95, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.17-8.14 (m, 3 H), 8.11-7.80 (m, 4 H), 7.93 (s, 1 H), 7.66-7.13 (m, 14 H), 5.60 (brs, 1 H), 5.44 (brs, 1 H), 5.71 (d, 1 H, J = 3.7 Hz), 5.08 (m, 1 H), 4.90 (dd, 1 H, J = 3.6, 12.1 Hz), 4.72 (dd, 1 H, J = 5.8, 11.8 Hz), 2.66-2.56 (m, 4 H), 1.81-1.63 (m, 4 H), 1.44-1.27 (m, 8 H), 0.97-0.88 (m, 6 H); 13 CNMR(75MHz, CDCl 3 ): 175.16, 172.07, 171.02, 166.12, 165.77, 165.22, 152.69, 151.06, 150.89, 147.74, 136.40, 133.73, 133.18, 129.97, 129.86, 129.62, 129.48, 128.92, 128.68, 128.59, 128.34, 124.70, 123.47, 122.35, 121.60, 120.68, 107.86, 82.96, 81.99, 77.70, 63.84, 34.34, 33.84, 31.21, 31.09, 24.55, 24.26, 22.23, 22.15, 13.82, 13.76; EI-MS (m/z): 445, 368, 270, 105, 99, 77, 43. Anal. Calcd for C 53 H 50 O 14 : C, 69.88%, H, 5.53%; Found: C, 69.63%, H, 5.80%. 6.4 4 -O-Hexanoyl daidzein 7-O-2,3,4,6 -tetra-o-benzoyl- -D-glucopyranoside (22) Compound 22 was obtained as a white foam: [ ] 20 D = 15.8 (c 0.93, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.11 (d, 1 H, J = 9.6 Hz), 8.03-7.86 (m, 8 H), 7.81 (s, 1 H), 7.59-7.30 (m, 14 H), 7.16 (d, 2 H, J = 8.5 Hz), 7.06-7.03 (m, 2 H), 6.04 (t, 1 H, J =9.2Hz), 5.85 (dd, 1 H, J = 7.4, 9.3 Hz), 5.74 (t, 1 H, J = 9.3 Hz), 5.60 (d, 1 H, J =7.4 Hz), 4.73 (dd, 1H, J = 2.4, 9.9 Hz), 4.53 (dd, 1 H, J = 6.9, 12.0 Hz), 4.41 (m, 1 H), 2.57 (t, 2 H, J = 7.6 Hz), 1.77 (m, 2 H), 1.40 (m, 4 H), 0.94 (t, 3 H, J =6.9Hz). 13 CNMR(75MHz, CDCl 3 ): 175.24, 172.25, 165.95, 165.68, 165.24, 165.00, 160.52, 157.20, 152.69, 150.76, 133.66, 133.52, 133.43, 133.29, 129.95, 129.89, 129.80, 129.36, 129.16, 128.81, 128.48, 128.37, 128.08, 124.53, 121.68, 120.09, 115.50, 104.43, 98.60, 73.01, 72.53, 71.48, 69.34, 63.07, 34.35, 31.24, 24.59, 22.29, 13.89. EI-MS (m/z): 579, 352, 254, 105, 77, 71, 43. Anal. Calcd for C 55 H 40 O 14 : C, 70.96%, H, 4.98%; Found: C, 71.00%, H, 5.34%. 6.5 4 -O-Hexanoyl daidzein 7-O-2,3,5 -tri-o-benzoyl- -D-arabinofuranoside (23) 10

Compound 23 was obtained as a white foam: [ ] 20 D = 48.6 (c 1.19, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 8.26 (d, 1 H, J = 9.6 Hz), 8.14-8.03 (m, 6 H), 7.97 (s, 1 H), 7.64-7.32 (m, 11 H), 7.24 (dd, 2 H, J = 1.5, 11.1 Hz), 7.17-7.14 (m, 3 H), 6.09 (brs, 1 H), 5.84 (brs, 1 H), 5.73 (d, 1 H, J = 3.9 Hz), 4.83 (dd, 1 H, J = 5.4, 8.1 Hz), 4.78-4.74 (m, 2 H), 2.57 (t, 2 H, J = 7.5 Hz), 1.79-1.74 (m, 2 H), 1.42-1.37 (m, 4 H), 0.93 (t, 3 H, J = 6.9 Hz). 13 C NMR (75 MHz, CDCl 3 ): 175.47, 172.24, 166.13, 165.68, 165.38, 160.08, 157.47, 152.87, 150.71, 133.79, 133.17, 129.95, 129.77, 129.28, 128.87, 128.62, 128.37, 128.07, 124.58, 121.65, 115.81, 103.88, 82.77, 81.95, 63.42, 34.37, 31.24, 24.59, 22.29, 13.90. EI-MS (m/z): 445, 253, 105, 77, 43. Anal. Calcd for C 47 H 40 O 12 : C, 70.85%, H, 5.06%; Found: C 70.80%, H, 4.89%. 6.6 3,3,4,5-Tetra-O-acetyl quercetin 7-O-2,3,4,6 -tetra-o-benzoyl- -Dglucopyranoside (24) Compound 24 was obtained as a white solid: [ ] 20 D = 23.9 (c 1.26, CHCl 3 ); 1 HNMR (300 MHz, CDCl 3 ): 7.97-7.87 (m, 8 H), 7.60-7.24 (m, 15 H), 6.99 (d, 1 H, J =2.4 Hz), 6.75 (d, 1 H, J = 2.7 Hz), 6.02 (t, 1 H, J = 9.3 Hz), 5.85 (dd, 1 H, J = 7.5, 9.0 Hz), 5.75 (t, 1 H, J = 9.0 Hz), 5.62 (d, 1 H, J = 7.5 Hz), 4.78 (m, 1 H), 4.49 (m, 2 H), 2.36 (s,3h),2.35(s,3h),2.34(s,6h). 13 CNMR(75MHz,CDCl 3 ): 169.82, 169.24, 167.82, 167.73, 167.67, 166.01, 165.60, 165.16, 164.92, 160.24, 157.47, 153.34, 150.87, 144.26, 142.12, 133.85, 133.56, 133.40, 133.32, 129.83, 129.83, 129.75, 129.45, 128.46, 128.36, 127.74, 126.25, 123.82, 123.70, 112.68, 109.72, 102.39, 98.20, 73.02, 72.39, 71.28, 69.20, 20.95, 20.60, 20.45. EI-MS (m/z): 470, 428, 386, 344, 302, 105, 77, 43. Anal. Calcd for C 57 H 44 O 20 H 2 O: C, 64.16%, H, 4.34%; Found: C, 64.30%, H, 4.30%. 7. General procedure for the deprotection of isoflavone glycosides The isoflavone ester (0.1 mmol) was dissolved in methanol-thf (4 ml, 1:1) followed by addition of K 2 CO 3 (2.2 equiv.) in H 2 O (0.3 ml). After stirring for 4 h at 40 o C, the mixture was neutralized with Dowex-50 H +, and then filtrated and concentrated. The 11

residue was purified by a silica gel column chromatography to afford the corresponding isoflavone glycoside. 7.1 4,8-Dihydroxy isoflavone 7-O- -D-arabinofuranoside (1) Compound 1 was obtained as a white solid: [ ] 14 24 D = 144.6 (c 1.12, MeOH), Lit. [ ] D = 125 (c 0.12, DMSO) (Ref. 3a); 1 H NMR (300 MHz, CD 3 OD-CDCl 3, 1:1): 8.15 (s, 1 H),7.73(d,1H,J = 9.0 Hz), 7.40 (d, 2 H, J = 8.1 Hz), 7.33 (d, 1 H, J = 9.0 Hz), 6.91 (d, 2 H, J = 8.1 Hz), 5.79 (brs, 1 H), 4.40 (brs, 1 H), 4.27 (m, 1 H), 4.18 (d, 1 H, J =3.6 Hz), 3.82 (dd, 1 H, J = 4.2, 11.7 Hz), 3.75 (dd, 1 H, J = 2.7, 12.0 Hz). 13 CNMR(75 MHz, DMSO-d 6 ): 175.35, 157.38, 157.21, 147.80, 146.21, 136.27, 130.29, 123.37, 122.63, 119.76, 115.13, 114.60, 107.33, 86.06, 81.57, 76.84, 61.30. ESI-MS (m/z): 403.1 [M + H + ]. 7.2 Daidzein 7-O- -D-glucopyranoside (2) Compound 2 was obtained as a white solid: [ ] 14 D = -23.0 (c 0.29, DMF); Lit. [ ] 24 D = -24.1 (c 0.001, DMSO) (Ref.: Lewis P.; Kaltia, S.; Wähälä, K. J. Chem. Soc. Perkin Trans. 1, 1998, 2481); 1 H NMR (300MHz, DMSO-d 6 ): 9.54 (brs, 1 H), 8.38 (s, 1 H), 8.03 (d, 1 H, J = 9.0 Hz), 7.39 (d, 2 H, J = 8.1 Hz), 7.22 (d, 2 H, J = 1.5 Hz), 7.13 (dd, 1H,J = 1.5, 9.0 Hz), 6.80 (d, 2 H, J = 8.1 Hz), 5.43 (d, 1 H, J = 2.4 Hz), 5.14 (d, 1 H, J =2.4Hz),5.08(m,2H),4.60(m,1H),4.08(m,1H),3.45(m,2H). 13 CNMR(75 MHz, DMSO-d 6 ): 174.91, 161.59, 157.44, 157.20, 153.48, 130.23, 127.12, 123.89, 122.47, 118.65, 115.75, 115.16, 103.58, 100.20, 77.38, 76.66, 73.32, 69.83, 60.83. ESI-MS (m/z): 439.3 [M + Na + ], 417.3 [M + H + ]. 7.3 Quercetin 7-O- -D-glucopyranoside (3) To a solution of 24 (145 mg, 0.14 mmol) in CHCl 3 (3mL)wasaddedasolutionof NaOMe in methanol (1 N, 5 ml) in one portion at argon atmosphere. The reaction was completed as shown by TLC after stirring at 6 o C for 4 h. The mixture was neutralized by Dowex-50 H + resin, and then filtrated. The organic phase was concentrated to give a residue, which was purified by a Sephadex LH-20 gel column (MeOH-H 2 O, 74:26) to 12

produce 3 as a yellow powder (32 mg, 50%): [ ] 14 D = -67.5 (c 0.26,MeOH),Lit. [ ] 20 D =-59(Ref.:Shelyuto,V.L.;Bondarenko,V.G.;Chem. Nat. Compd. 1985, 21, 534); 1 H NMR (300 MHz, DMSO-d 6 ): 12.50 (s, 1 H), 9.50 (brs, 3 H), 7.72 (d, 1 H, J = 2.2 Hz), 7.55 (d, 1 H, J = 8.8 Hz), 6.90 (d, 1 H, J = 8.5 Hz), 6.76 (d, 1H, J =1.9Hz), 6.42 (d, 1 H, J = 1.7 Hz), 5.19 (brs, 1 H), 5.07 (d, 1 H, J = 7.1 Hz), 5.09 (brs, 2 H), 4.60 (brs, 1 H), 3.70 (m, 2 H), 3.50-3.20 (m, 4 H). 13 C NMR (75 MHz, DMSO-d 6 ): 176.18, 162.89, 160.55, 155.94, 148.08, 147.77, 145.24, 136.27, 122.02, 120.26, 115.78, 104.85, 100.10, 98.96, 94.49, 77.35, 76.56, 73.32, 69.75, 60.81. ESI-MS (m/z): 465.2 [M + H + ]. 7.4 4, 8-Dihydroxyisoflavone 7-O- -D-glucopyranoside (25) Compound 25 was obtained as a white foam: [ ] 14 24 D =-20.8(c 0.17, DMF), Lit. [ ] D = -55.9 (c 0.55, MeOH)(Ref.3b); 1 H NMR (300 MHz, DMSO-d 6 ): 9.55 (brs, 1 H), 8.39 (s, 1 H), 7.80 (d, 2 H, J = 8.4 Hz), 7.49 (d, 1 H, J = 9.3 Hz), 7.39 (d, 2 H, J =8.4 Hz), 7.28 (d, 2 H, J = 8.7 Hz), 5.09 (brs, 1 H, OH), 5.08 (d, 1 H, J = 3.6 Hz), 4.89 (d, 1 H, J = 7.2 Hz), 4.61 (brs, 1 H), 3.72 (brd, 1 H, J =11.1Hz),3.48(m,1H). 13 CNMR (75 MHz, DMSO-d 6 ): 175.34, 157.38, 153.39, 148.79, 146.07, 135.88, 130.24, 123.29, 122.63, 119.88, 115.13, 114.74, 113.87, 102.08, 77.48, 75.87, 73.42, 69.89, 60.83. ESI-MS (m/z): 433.2 [M + H + ], 432.3 [M + ]. 7.5 4, 8-Dihydroxyisoflavone 7-O- -L-rhamnopyranoside (26) Compound 26 was obtained as a white foam: [ ] 14 D =-72.4(c 0.61, MeOH); 1 HNMR (300 MHz, DMSO-d 6 ): 9.72 (brs, 1 H), 9.54 (brs, 1 H), 8.40 (s, 1 H), 7.55 (d, 2 H, J = 9.0 Hz), 7.40 (d, 1 H, J = 8.7 Hz), 7.26 (d, 1 H, J = 9.3 Hz), 6.81 (d, 2 H, J =8.1Hz), 5.47 (brs,1 H, OH), 5.06 (brs, 1 H), 4.90 (d, 1 H, J = 5.1 Hz), 4.67 (d, 1 H, J =5.4Hz), 3.98 (brs, 1 H), 3.87 (m, 1 H), 3.55 (m, 1 H), 1.11 (d, 3 H, J =6.3Hz). 13 CNMR(75 MHz, DMSO-d 6 ): 175.35, 157.40, 153.22, 148.00, 146.53, 135.80, 130.28, 123.39, 122.63, 119.65, 115.30, 115.14, 114.25, 99.74, 72.03, 70.25, 70.10, 69.98, 18.00. ESI- MS (m/z): 417.1 [M + H + ]. HRMS: Calcd for C 21 H 20 O 9 : 439.1014 [M + Na + ]. Found: 439.1000. 13

7.6 Daidzein 7-O- -D-arabinofuranoside (27) Compound 27 was obtained as a white solid: [ ] 14 D = 153.8 (c 0.22, MeOH); 1 HNMR (300 MHz, 1:1 CD 3 OD-CDCl 3 ): 8.16 (d, 1 H, J = 9.3 Hz), 8.08 (s, 1 H), 7.39 (d, 2 H, J = 8.7 Hz), 7.22 (d, 1 H, J = 2.4 Hz), 7.17 (dd, 1 H, J = 8.7, 2.4 Hz), 6.90 (d, 2 H, J = 8.1 Hz), 5.72 (brs, 1 H), 4.36 (brs, 1 H), 4.17 (m, 1 H), 4.11 (m, 1 H), 3.85 (dd, 1 H, J = 3.0, 12.0 Hz), 3.74 (dd, 1 H, J = 4.2, 12.0 Hz). 13 CNMR(75MHz,1:1CD 3 OD- CDCl 3 ): 177.88, 162.40, 158.76, 158.09, 154.16, 131.10, 128.14, 125.93, 123.71, 119.60, 116.96, 107.52, 104.41, 86.92, 82.89, 77.88, 62.31. ESI-MS (m/z): 409.1 [M + Na + ], 387.1 [M + H + ]. HRMS: Calcd for C 20 H 18 O 8 : 409.0902 [M + Na + ]. Found: 409.0894. 14