Supplementary Figure 1. 1 H and 13 C NMR spectra for compound S2.

Transcription

1 Supplementary Figure 1. 1 H and 13 C NMR spectra for compound S2. 1

2 Supplementary Figure 2. 1 H and 13 C NMR spectra for compound S3. 2

3 Supplementary Figure 3. 1 H and 13 C NMR spectra for compound S4. 3

4 Supplementary Figure 4. 1 H and 13 C NMR spectra for compound S5. 4

5 Supplementary Figure 5. 1 H and 13 C NMR spectra for compound S6. 5

6 Supplementary Figure 6. 1 H and 13 C NMR spectra for compound S7. 6

7 Supplementary Figure 7. 1 H and 13 C NMR spectra for compound 4α. 7

8 Supplementary Figure 8. 1 H and 13 C NMR spectra for compound 4β. 8

9 Supplementary Figure 9. 1 H and 13 C NMR spectra for compound 12. 9

10 Supplementary Figure H and 13 C NMR spectra for compound

11 Supplementary Figure H and 13 C NMR spectra for compound

12 Supplementary Figure H and 13 C NMR spectra for compound S8. 12

13 Supplementary Figure H and 13 C NMR spectra for compound

14 Supplementary Figure H and 13 C NMR spectra for compound

15 Supplementary Figure H and 13 C NMR spectra for compound

16 Supplementary Figure H and 13 C NMR spectra for compound

17 Supplementary Figure H and 13 C NMR spectra for compound S2. 1

18 Supplementary Figure H and 13 C NMR spectra for compound 5. 18

19 Supplementary Figure H and 13 C NMR spectra for compound

20 Supplementary Figure H and 13 C NMR spectra for compound 2β. 20

21 Supplementary Figure H and 13 C NMR spectra for compound S10. 21

22 Supplementary Figure H and 13 C NMR spectra for compound

23 Supplementary Figure H and 13 C NMR spectra for compound S12. 23

24 Supplementary Figure H and 13 C NMR spectra for compound

25 Supplementary Figure H and 13 C NMR spectra for compound

26 Supplementary Figure H and 13 C NMR spectra for compound

27 Supplementary Figure H and 13 C NMR spectra for compound

28 Supplementary Figure H and 13 C NMR spectra for compound

29 Supplementary Figure H and 13 C NMR spectra for compound 9. 29

30 Supplementary Figure H and 13 C NMR spectra for compound

31 Supplementary Figure H and 13 C NMR spectra for compound 7. 31

32 Supplementary Figure H and 13 C NMR spectra for compound S14. 32

33 Supplementary Figure H and 13 C NMR spectra for compound

34 Supplementary Figure H and 13 C NMR spectra for compound

35 Supplementary Figure H and 13 C NMR spectra for compound 28α. 35

36 Supplementary Figure H and 13 C NMR spectra for compound S15. 36

37 Supplementary Figure H and 13 C NMR spectra for compound

38 Supplementary Figure H and 13 C NMR spectra for compound

39 Supplementary Figure H and 13 C NMR spectra for compound

40 Supplementary Figure H and 13 C NMR spectra for compound S17. 40

41 Supplementary Figure H and 13 C NMR spectra for compound S18. 41

42 Supplementary Figure H and 13 C NMR spectra for compound S19. 42

43 Supplementary Figure H and 13 C NMR spectra for compound

44 Supplementary Figure H and 13 C NMR spectra for compound

45 Supplementary Figure H and 13 C NMR spectra for compound

46 Supplementary Figure H and 13 C NMR spectra for compound

47 Supplementary Figure H and 13 C NMR spectra for compound

48 Supplementary Figure H and 13 C NMR spectra for compound

49 Supplementary Figure H and 13 C NMR spectra for compound

50 Supplementary Figure H and 13 C NMR spectra for compound

51 Supplementary Figure H and 13 C NMR spectra for compound S20. 51

52 Supplementary Figure H and 13 C NMR spectra for compound 3. 52

53 Supplementary Figure H and 13 C NMR spectra for compound

54 Supplementary Figure H and 13 C NMR spectra for compound 40α. 54

55 Supplementary Figure H and 13 C NMR spectra for compound 1. 55

56 Supplementary Figure H and 13 C NMR spectra for compound

57 Supplementary Figure H and 13 C NMR spectra for compound

58 Supplementary Figure H and 13 C NMR spectra for compound

59 Supplementary Figure 59. X-ray structure of S14. CCDC contains the supplementary crystallographic data for S14 that is available free of charge from The Cambridge Crystallographic Data Centre via 59

60 Supplementary Figure 60. Comparison of the 1 H NMR spectra of the synthetic 1, 31, and an authentic periploside A. (The authentic sample of periploside A was provided by Prof. Weimin Zhao from Shanghai Institute of Materia Medica, Chinese Academy of Sciences.) 60

61 Supplementary Figure 61. Comparison of the 13 C NMR spectra of the synthetic 1, 31, and an authentic periploside A. 61

62 Inhibition Rate % Log 10 (compound) ng/ml Supplementary Figure 62. Inhibition of the growth of T-lymphocytes by synthetic compounds 1, 31, and Cytotoxicity % Log 10 (compound) ng/ml Supplementary Figure 63. Inhibition of concanavalin A (ConA)-induced T-lymphocyte proliferation by synthetic compounds 1, 31, and

63 Supplementary Table 1. Attempts at synthesis of the acetal glycoside 24. Entry Promoter Temp/ o C 10/11 a Results: NBS/TfOH -50~0 1:1 80% 8% 2 Tf 2 O,BSP,TTBP -60~rt 1:1 70% 7% MeOTf,DTBMP 0~rt no reaction NIS/TfOH -40~-30 1:1 65% 31% NIS/TfOH -40~-30 1:2 41% 41% NIS/TfOH 0 1:2 complex NIS/AgOTf 0 1:2 complex 8 DMTST,TTBP 0 1:1.8 90% trace 9 b b NIS/TfOH -30 1:2 (inverse procedure) 20% 50% NIS/TfOH -40~-30 1:5 36% 63% NIS/TfOH -30 1:5 (inverse procedure) 25% 75% a The mole ratio of 10 and 11. b Reaction was conducted by inverse procedure 1. 63

64 Supplementary Table 2. Attempts at the synthesis of the FABO disaccharide 37. Entry Promoter Temp/ o C Solvent Results: NIS/TMSOTf -40 ~ -30 CH 2 Cl 2 90% 0 2 MeOTf/TTBP -72 ~ rt CH 2 Cl 2 80% 0 3 DMTST/TTBP -50 ~ rt CH 2 Cl 2 90% a b 9 10 c 11 c CuBr 2 /Bu 4 NBr 0 ~ rt CH 2 Cl 2 90% 0 MeOTf/DTBMP (frozen condition) BSP/Tf 2 O/TTBP p-xylene 80% 10 CH 2 Cl 2 95% 0 BSP/Tf 2 O/DTBP -78 CH 2 Cl 2 80% 10 BSP/Tf 2 O/DTBP -78 CH 2 Cl 2 no reaction (inverse procedure) BSP/Tf 2 O/DTBP -100 CH 2 Cl 2 /Et 2 O 57% 41% BSP/Tf 2 O/DTBP BSP/Tf 2 O/DTBP -114 CH 2 Cl 2 /Et 2 O 35% 30% -114 Et 2 O 18% 64% a Reaction was conducted at frozen condition 2. b Reaction was conducted by inverse procedure 1. c 38 was isolaterd asasingle diastereoisomer. Supplementary Table 3. Comparison of the 13 C NMR (500 MHz, CDCl 3 ) spectroscopic data of the synthetic 1 and C1 -epi-periploside A (31) with those reported for the natural periploside A 3. 64

65 Position aglycone Natural Synthetic 1 C1 -epi-perip periploside A loside A (31) Position cym. Natural Synthetic1 C1 -epi-perip periploside A loside A (31) C C C C C C C C C C C C C OMe C cym. C C C C C C C C C C C C C OMe C cym. C C C C C C C C C C can. C C OMe C dig. C C C C C C C C OCH 2 O C ole. C C OMe C OAc C C C C OMe

66 Supplementary Methods General Remarks. All reactions were carried out under nitrogen or argon with anhydrous solvents in flame-dried glassware, unless otherwise noted. All the glycosylation reactions were performed in the presence of 4Å or 5Å molecular sieves, which were flame-dried immediately before use in the reaction under high vacuum. Glycosylation solvents were dried using a solvent purification system and used directly without further drying. The chemicals used were reagent grade as supplied, except where noted. Analytical thin-layer chromatography was performed using silica gel 60 F254 glass plates. Compound spots were visualized by UV light (254 nm) or by heating with a solution with 10% H 2 SO 4 in ethanol. Flash column chromatography was performed on silica gel. NMR spectra were recorded on Bruker AV-400 or Agilent 500 instrument and referenced using Me 4 Si (0 ppm), residual CHCl 3 ( 1 H NMR δ = 7.26 ppm, 13 C NMR δ = ppm). Splitting patterns are indicated as s (singlet), d (doublet), t (triplet), q (quartet), and brs (broad singlet) for 1 H NMR data. ESI-MS and MALDI-MS were run on an IonSpec Ultra instrument using HP5989A or VG Quattro MS. Optical rotations were measured using a Perkin-Elmer 241 polarimeter. Synthesis of digitalosyl ortho-cyclopropylethynylbenzoate 4. p-methoxyphenyl 2-O-benzyl-3,4-O-isopropylidene-α-D-fucopyranoside (S2) To a solution of p-methoxyphenyl α-d-fucopyranoside (S1) 4 (3.74 g, 13.8 mmol) in acetone (100 ml) was added 2,2-dimethoxypropane (2.90 ml, 23.5 mmol). The mixture was cooled to 0 C, then BF 3 OEt 2 (0.26 ml, 2.08 mmol) was added dropwise. The mixture was stirred for 12 h at rt before being quenched by addition of Et 3 N (2 ml). The mixture was concentrated in vacuo. The residue was purified by flash chromatography (petroleum ether/etoac = 4:1) to afford a colorless syrup (

67 g, 92%). The above syrup (3.95 g, 12.7 mmol) was dissolved in DMF (50 ml) and cooled to 0 C. Benzyl bromide (3.28 ml, 27.7 mmol) and sodium hydride (60% in oil) (1.1 g, 27.7 mmol) were added carefully. The mixture was stirred for 3 h while warming to rt. The solution was quenched with water and diluted with CH 2 Cl 2. The water phase was extracted with CH 2 Cl 2 for three times. The combined organic layer was washed with brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 20:1) to afford S2 (4.7 g, 93%) as a white solid: [α] 28 D = (c = 1.7, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 5H), 7.02 (d, J = 9.0 Hz, 2H), 6.82 (d, J = 9.0 Hz, 2H), 5.33 (d, J = 3.3 Hz, 1H), (m, 2H), 4.49 (dd, J = 7.6, 5.7 Hz, 1H), (m, 1H), 4.10 (dd, J = 5.3, 2.3 Hz, 1H), 3.76 (s, 3H), 3.64 (dd, J = 7.8, 3.4 Hz, 1H), 1.43 (s, 3H), 1.37 (s, 3H), 1.29 (d, J = 6.7 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 155.0, 151.1, 138.3, 128.5, 128.0, 127.8, 117.8, 114.7, 109.0, 96.5, 76.2, 76.2, 76.0, 72.4, 64.0, 55.7, 28.3, 26.5, 16.4; HRMS (ESI) calcd for C 23 H 28 O 6 Na [M+Na] , found p-methoxyphenyl 2-O-benzyl-α-D-fucopyranoside (S3) To a solution of S2 (3.94 g, 9.84 mmol) in methanol (50 ml) and water (5 ml) was added an aqueous solution of 5% HCl (a few drops) until ph = 3. The mixture was heated to 50 C and stirred for 10 h until the TLC showed complete conversion. The mixture was quenched with a saturated NaHCO 3 solution. The solvent was then removed. The residue was diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, respectively, and was then dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 2:1) to afford S3 (3.0 g, 85%) as a white solid: [α] 29 D = (c = 1.4, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 5H), 7.00 (d, J = 9.1 Hz, 2H), 6.82 (d, J = 9.1 Hz, 2H), 5.39 (d, J = 3.5 Hz, 1H), (m, 2H), 4.19 (dt, J = 9.8, 3.1 Hz, 1H), 4.09 (q, J = 6.5 Hz, 1H), (m, 2H), 3.77 (s, 3H), 2.94 (d, J = 3.1 Hz, 1H), 2.72 (d, 67

68 J = 1.8 Hz, 1H), 1.25 (d, J = 6.6 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 155.1, 151.3, 137.8, 128.7, 128.2, 118.1, 114.7, 96.3, 76.3, 72.6, 71.6, 69.5, 66.4, 55.8, 16.2; HRMS (ESI) calcd for C 20 H 24 O 6 Na [M+Na] , found p-methoxyphenyl 2-O-benzyl-3-O-methyl-α-D-fucopyranoside (S4) A solution of S3 (2.25 g, 6.25 mmol) in methanol (20 ml) was refluxed in the presence of dibutyltin oxide (1.87 mg, 7.50 mmol) for 4 h. The solvent was then removed in vacuo, and the residue was dried azeotropically with toluene and dissolved in DMF (20 ml). Methyl iodide (1.9 ml, 31.3 mmol) and CsF (1.4 g, 9.37 mmol) were added, and the mixture was stirred at rt for 12 h. The foamy solution was diluted with 10% HCl and extracted with EtOAc. The combined organic layer was washed with brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 3:1) to afford S4 (2.3 g, 98%) as a syrup: [α] 28 D = (c = 1.3, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 5H), 7.01 (d, J = 9.1 Hz, 2H), 6.81 (d, J = 9.1 Hz, 2H), 5.31 (d, J = 3.5 Hz, 1H), 4.78 (d, J = 12.1 Hz, 1H), 4.64 (d, J = 12.1 Hz, 1H), 4.07 (q, J = 6.4 Hz, 1H), 3.93 (d, J = 2.3 Hz, 1H), 3.87 (dd, J = 9.8, 3.6 Hz, 1H), 3.80 (dd, J = 9.8, 3.2 Hz, 1H), 3.74 (s, 3H), 3.57 (s, 3H), 2.65 (s, 1H), 1.26 (d, J = 6.6 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 154.9, 151.2, 138.4, 128.4, 127.8, 127.7, 118.1, 114.5, 97.0, 79.5, 75.2, 73.1, 69.1, 66.1, 58.0, 55.6, 16.2; HRMS (ESI) calcd for C 21 H 26 O 6 Na [M+Na] , found p-methoxyphenyl 2-O-benzyl-4-O-chloroacetyl-3-O-methyl-α-D-fucopyranoside (S5) To a solution of S4 (2.20 g, 5.89 mmol) in CH 2 Cl 2 (20 ml) were added Et 3 N (

69 ml, 17.7 mmol) and DMAP (72 mg, 0.59 mmol). The mixture was cooled to 0 C, (ClAc) 2 O (1.51 g, 8.83 mmol) was added under stirring. After stirring for 2 h at rt, the reaction was quenched with MeOH. The solution was diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, respectively, and was then dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 6:1 to 2:1) to afford S5 (2.16 g, 92% based on 81% conversion) as a colorless syrup and recovered S4 (260 mg, 11%). S5: [α] 28 D = (c = 1.8, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 5H), 6.91 (d, J = 9.0 Hz, 2H), 6.74 (d, J = 9.0 Hz, 2H), 5.36 (d, J = 2.8 Hz, 1H), 5.24 (d, J = 3.5 Hz, 1H), 4.74 (d, J = 12.1 Hz, 1H), 4.56 (d, J = 12.1 Hz, 1H), (m, 3H), 3.84 (dd, J = 10.0, 3.2 Hz, 1H), 3.71 (dd, J = 10.1, 3.6 Hz, 1H), 3.67 (s, 3H), 3.40 (s, 3H), 1.04 (d, J = 6.5 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 167.3, 155.1, 151.1, 138.4, 128.4, 127.9, 127.8, 118.2, 114.6, 97.4, 77.9, 75.0, 73.4, 72.4, 65.2, 58.1, 55.7, 40.8, 16.2; HRMS (ESI) calcd for C 23 H 27 O 7 ClNa [M+Na] , found p-methoxyphenyl 4-O-chloroacetyl-3-O-methyl-α-D-fucopyranoside (S6) To a solution of S5 (2.06 g, 4.57 mmol) in EtOAc (15 ml) was added palladium on carbon (10% Pd/C, 206 mg). The suspension was stirred under hydrogen pressure (1 atm) for 5 h and then filtered. The filtrate was concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 2:1) to afford S6 (1.62 g, 98%) as a white solid: [α] 28 D = (c = 1.2, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.01 (d, J = 9.1 Hz, 2H), 6.83 (d, J = 9.1 Hz, 2H), 5.46 (d, J = 3.8 Hz, 1H), 5.44 (d, J = 2.8 Hz, 1H), 4.23 (dd, J = 13.1, 6.6 Hz, 1H), 4.18 (s, 2H), 3.97 (ddd, J = 10.4, 7.0, 3.8 Hz, 1H), 3.76 (s, 3H), 3.71 (dd, J = 10.0, 3.2 Hz, 1H), 3.47 (s, 3H), 2.45 (d, J = 7.2 Hz, 1H), 1.16 (d, J = 6.6 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 167.4, 155.4, 150.8, 118.2, 114.8, 98.5, 78.7, 71.5, 68.1, 65.7, 57.9, 55.8, 40.8, 16.3; HRMS (ESI) calcd for C 16 H 21 O 7 ClNa [M+Na] , found

70 p-methoxyphenyl 2-O-acetyl-4-O-chloroacetyl-3-O-methyl-α-D-fucopyranoside (S7) To a solution of S6 (530 mg, 1.47 mmol) in CH 2 Cl 2 (10 ml) were added Et 3 N (0.61 ml, 4.41 mmol) and DMAP (18 mg, 0.15 mmol). The mixture was cooled to 20 C, then Ac 2 O (0.17 ml, 1.76 mmol) was added under stirring. The mixture was stirred for 1 h while warming to 10 C. The reaction was quenched with MeOH. The solution was diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, respectively. The organic phase was dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 6:1) to afford S7 (570 mg, 96%) as a white solid: [α] 25 D = (c = 1.1, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.97 (d, J = 9.1 Hz, 2H), 6.82 (d, J = 9.1 Hz, 2H), 5.55 (d, J = 3.7 Hz, 1H), 5.48 (d, J = 2.8 Hz, 1H), 5.09 (dd, J = 10.5, 3.7 Hz, 1H), 4.25 (dd, J = 13.1, 6.5 Hz, 1H), 4.20 (s, 2H), 3.93 (dd, J = 10.5, 3.3 Hz, 1H), 3.76 (s, 3H), 3.44 (s, 3H), 2.10 (s, 3H), 1.17 (d, J = 6.5 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.6, 167.4, 155.4, 150.9, 118.1, 114.8, 96.2, 75.6, 71.9, 69.7, 65.3, 58.1, 55.8, 40.8, 21.0, 16.2; HRMS (ESI) calcd for C 18 H 23 O 8 ClNa [M+Na] , found Digitalosyl ortho-cyclopropylethynylbenzoate 4 To a solution of S7 (1.20 g, 2.98 mmol) in toluene/ch 3 CN/H 2 O (6 ml/9 ml/6 ml) was added CAN (8.20 g, 14.9 mmol). After stirring for 1 h at rt, the solution was poured into ice water and extracted with CH 2 Cl 2. The organic layer was washed with saturated aqueous NaHCO 3 and brine, respectively, and was then dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1.5:1) to afford the corresponding hemiacetal (810 mg, 91%) as a 70

71 colorless syrup. To a solution of the above hemiacetal (367 mg, 1.24 mmol) in CH 2 Cl 2 (5 ml) were added 2-(cyclopropylethynyl)benzoic acid (345 mg, 1.86 mmol) 5, DMAP (227 mg, 1.86 mmol), EDCI (428 mg, 2.23 mmol), and 5 Å MS (500 mg). The mixture was stirred at rt for 3.5 h, and then filtered. The filtrate was diluted with CH 2 Cl 2, washed with saturated aqueous NaHCO 3 and brine, respectively. The organic phase was dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 5:1) to afford 4 (559 mg, 97%, β/α = 1.6/1) as a syrup. 4α: [α] 26 D = (c = 0.9, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.92 (d, J = 7.8 Hz, 1H), (m, 2H), (m, 1H), 6.58 (d, J = 3.6 Hz, 1H), 5.53 (d, J = 2.5 Hz, 1H), 5.25 (dd, J = 10.5, 3.7 Hz, 1H), 4.45 (q, J = 6.4 Hz, 1H), 4.22 (s, 2H), 3.93 (dd, J = 10.5, 3.2 Hz, 1H), 3.41 (s, 3H), 2.04 (s, 3H), (m, 1H), 1.22 (d, J = 6.5 Hz, 3H), (m, 4H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.3, 167.5, 164.6, 135.0, 132.2, 131.0, 130.8, 127.4, 124.8, 99.5, 91.1, 75.7, 75.1, 71.5, 68.5, 67.9, 57.9, 40.8, 20.9, 16.4, 9.1, 9.1, 0.8; HRMS (ESI) calcd for C 23 H 25 O 8 ClNa [M+Na] , found β: [α] 26 D = 7.0 (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.95 (d, J = 7.8 Hz, 1H), (m, 2H), (m, 1H), 5.85 (d, J = 8.4 Hz, 1H), 5.45 (d, J = 3.1 Hz, 1H), 5.31 (dd, J = 9.9, 8.6 Hz, 1H), 4.24 (d, J = 0.9 Hz, 2H), 3.97 (q, J = 6.3 Hz, 1H), 3.50 (dd, J = 10.1, 3.4 Hz, 1H), 3.40 (s, 3H), 2.01 (s, 3H), (m, 1H), 1.28 (d, J = 6.4 Hz, 3H), 0.90 (d, J = 6.7 Hz, 4H); 13 C NMR (100 MHz, CDCl 3 ) δ 169.9, 167.5, 163.7, 134.6, 132.6, 131.1, 129.5, 127.3, 125.9, 100.5, 92.7, 80.0, 74.5, 70.7, 70.4, 69.5, 58.4, 40.9, 20.9, 16.3, 9.1, 9.1, 0.9; HRMS (ESI) calcd for C 23 H 25 O 8 ClNa [M+Na] , found Synthesis of tetrasaccharide donor 2 p-methoxyphenyl 3,4-di-O-tert-butyldiphenylsilyl-2,6-dideoxy-β-D-ribo-hexopyranoside (12) 71

72 To a solution of 8 6 (4.05 g, 5.10 mmol) and p-methoxyphenol (MpOH) (1.27 g, 10.2 mmol) in toluene (10 ml) was added 4 Å MS (5.0 g) at rt. After stirring for 30 min at 40 o C, PPh 3 AuNTf 2 (377 mg, 0.51 mmol) was added to the mixture. After stirring for another 2 h at this temperature, Et 3 N was added to quench the reaction. The resulting mixture was filtered and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 80:1) to afford 12 (3.54 g, 95%) as a white foam: [α] 26 D = 31.7 (c = 2.6, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 8H), (m, 10H), (m, 2H), 6.77 (dd, J = 21.6, 9.1 Hz, 4H), 5.43 (brs, 1H), (brs, 1H), (m, 1H), 3.72 (s, 3H), 3.63 (brs, 1H), 2.34 (brs, 1H), (m, 1H), (m, 18H), 0.80 (brs, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 154.6, 151.5, 136.3, 136.2, 136.2, 134.0, 133.9, 133.6, 129.9, 129.8, 129.7, 127.8, 127.7, 127.7, 117.6, 114.4, 97.5, 75.5, 55.7, 27.4, 27.3, 19.6, 19.5, 19.0; HRMS (ESI) calcd for C 45 H 54 O 5 Si 2 Na [M+Na] , found p-methoxyphenyl 3-O-acetyl-2,6-dideoxy-β-D-ribo-hexopyranoside (13) To a solution of 12 (1.68 g, 2.30 mmol) in THF (10 ml) was added TBAF (1 M in THF, 9.2 ml, 9.2 mmol) at 0 o C. After stirring for 14 h at rt, the solution was diluted with CH 2 Cl 2. The mixture was washed with water and brine, respectively, and was then dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1.5:1) to afford a syrup (562 mg, 99%). The above syrup (230 mg, 0.90 mmol) was dissolved in CH 2 Cl 2 (3 ml). Trimethylorthoacetate (0.35 ml, 2.71 mmol) and a catalytic amount of p-toluenesulfonic acid (7.8 mg, mmol) were added at rt. The mixture was stirred for 3 h, and TLC indicated that the starting material was consumed. The 72

73 solvent was removed and the residue was dissolved in THF/H 2 O (4 ml, 1:1). p-toluenesulfonic acid (78 mg, 0.45 mmol) was added, and the stirring was continued for 1 h. The reaction was quenched with saturated NaHCO 3 solution. The mixture was extracted with CH 2 Cl 2. The combined organic layer was washed with brine, dried over Na 2 SO 4, and then concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 3:1) to afford 13 (248 mg, 93%) as a colorless syrup: [α] 27 D = +7.0 (c = 1.5, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.98 (d, J = 8.4 Hz, 2H), 6.82 (d, J = 8.6 Hz, 2H), 5.37 (d, J = 2.6 Hz, 1H), 5.27 (d, J = 9.1 Hz, 1H), (m, 1H), 3.77 (s, 3H), 3.53 (d, J = 3.6 Hz, 1H), 2.28 (d, J = 14.1 Hz, 1H), 2.19 (d, J = 6.1 Hz, 1H), 2.13 (s, 3H), (m, 1H), 1.36 (d, J = 6.1 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 171.3, 155.1, 151.3, 117.9, 114.6, 97.0, 72.1, 70.9, 70.8, 55.8, 35.5, 21.3, 18.3; HRMS (ESI) calcd for C 15 H 20 O 6 Na [M+Na] , found Disaccharide 14 To a solution of 8 (2.55 g, 3.22 mmol) and 13 (636 mg, 2.15 mmol) in toluene (15 ml) was added 4 Å MS at rt. After stirring for 30 min at 40 o C, PPh 3 AuNTf 2 (159 mg, mmol) was added to the mixture. After stirring for another 2 h at this temperature, Et 3 N was added to quench the reaction. The resulting mixture was filtered and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 12:1) to afford 14 (1.92 g, 99%) as a white foam: [α] 26 D = (c = 1.2, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 6H), 7.51 (d, J = 7.0 Hz, 2H), (m, 10H), (m, 2H), 6.95 (d, J = 8.7 Hz, 2H), 6.80 (d, J = 8.7 Hz, 2H), 5.21 (s, 1H), 5.16 (d, J = 8.7 Hz, 1H), 4.76 (d, J = 8.1 Hz, 1H), 4.33 (s, 1H), 4.09 (brs, 1H), (m, 1H), 3.75 (s, 3H), 3.42 (d, 1H), 3.03 (d, J = 7.6 Hz, 1H), 2.26 (d, J = 13.9 Hz, 1H), (m, 5H), (m, 1H), 1.18 (d, J = 6.1 Hz, 3H), 1.10 (s, 9H), 0.93 (s, 9H), 0.77 (d, J = 6.1 Hz, 3H); 13 C NMR (100 73

74 MHz, CDCl 3 ) δ 170.3, 155.1, 151.4, 136.3, 136.2, 136.2, 134.4, 134.0, 133.9, 133.7, 130.0, 129.8, 129.8, 127.8, 127.7, 127.7, 127.6, 117.9, 114.6, 97.1, 79.0, 75.9, 69.7, 68.9, 55.8, 35.3, 27.4, 27.3, 21.4, 19.8, 19.5, 18.6, 18.5; HRMS (ESI) calcd for C 53 H 66 O 9 Si 2 Na [M+Na] , found Disaccharide S8 To a solution of 14 (1.92 g, 2.13 mmol) in THF (8 ml) was added TBAF (1 M in THF, 13.6 ml, 13.6 mmol) at 0 o C. After stirring for 30 h at rt, the solution was diluted with CH 2 Cl 2. The mixture was washed with water and brine, dried over Na 2 SO 4, and then concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1.5:1) to afford S8 (863 mg, 94%) as a white foam: [α] 26 D = (c = 1.1, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.96 (d, J = 8.4 Hz, 2H), 6.81 (d, J = 8.5 Hz, 2H), 5.45 (d, J = 2.2 Hz, 1H), 5.23 (d, J = 8.9 Hz, 1H), 4.86 (d, J = 9.5 Hz, 1H), 4.09 (brs, 1H), (m, 1H), 3.76 (s, 3H), (m, 1H), 3.43 (d, J = 7.0 Hz, 1H), 3.27 (d, J = 8.0 Hz, 1H), 2.50 (brs, 1H), 2.30 (d, J = 14.2 Hz, 2H), (m, 5H), (m, 1H), 1.32 (d, J = 6.1 Hz, 3H), 1.24 (d, J = 5.7 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.4, 155.1, 151.4, 117.9, 114.6, 98.8, 97.2, 79.2, 72.9, 69.8, 69.7, 69.4, 68.2, 55.8, 37.8, 35.6, 21.4, 18.5, 18.1; HRMS (ESI) calcd for C 21 H 30 O 9 Na [M+Na] , found Disaccharide 15 The white foam S8 (863 mg, 2.02 mmol) was dissolved in CH 2 Cl 2 (5 ml). Trimethylorthoacetate (0.77 ml, 6.07 mmol) and a catalytic amount of p-toluenesulfonic acid (17 mg, 0.10 mmol) were added at rt. The mixture was stirred for 1 h, and TLC indicated that the starting material was consumed. The solvent was removed and the residue was dissolved in THF/H 2 O (10 ml, 1:1). p-toluenesulfonic 74

75 acid (170 mg, 1.01 mmol) was added, and the stirring was continued for 1 h. The reaction was quenched with saturated NaHCO 3 solution. The mixture was extracted with CH 2 Cl 2. The combined organic layer was washed with brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1:1) to afford 15 (866 mg, 91%) as a white foam: [α] 27 D = (c = 0.9, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.97 (d, J = 9.0 Hz, 2H), 6.81 (d, J = 9.1 Hz, 2H), 5.46 (d, J = 3.3 Hz, 1H), (m, 2H), 4.77 (dd, J = 9.6, 1.7 Hz, 1H), (m, 1H), 3.76 (s, 3H), (m, 1H), 3.43 (dd, J = 9.0, 3.1 Hz, 1H), (m, 1H), 2.31 (ddd, J = 14.1, 4.0, 2.2 Hz, 1H), 2.14 (s, 3H), (m, 4H), (m, 2H), (m, 1H), 1.32 (d, J = 6.3 Hz, 3H), 1.26 (d, J = 6.3 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 171.4, 170.3, 155.1, 151.4, 117.9, 114.7, 98.7, 97.2, 79.3, 72.2, 71.2, 70.4, 69.7, 69.5, 55.8, 36.0, 35.6, 21.4, 21.3, 18.5, 18.0; HRMS (ESI) calcd for C 23 H 32 O 10 Na [M+Na] , found Trisaccharide 16 To a solution of 8 (2.40 g, 3.03 mmol) and 15 (887 mg, 1.89 mmol) in toluene (15 ml) was added 4Å MS at rt. After stirring for 30 min at 40 o C, PPh 3 AuNTf 2 (140 mg, mmol) was added to the mixture. After stirring for another 2 h at this temperature, Et 3 N was added to quench the reaction. The resulting mixture was filtered and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 8:1) to afford 16 (2.02 g, 99%) as a white foam: [α] 25 D = (c = 2.2, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 6H), (m, 12H), 7.21 (t, J = 7.5 Hz, 2H), 6.98 (d, J = 9.1 Hz, 2H), 6.82 (d, J = 9.1 Hz, 2H), 5.48 (d, J = 3.1 Hz, 1H), 5.25 (dd, J = 9.1, 1.7 Hz, 1H), 5.13 (d, J = 2.8 Hz, 1H), 4.79 (d, J = 7.6 Hz, 1H), 4.66 (d, J = 9.4 Hz, 1H), 4.31 (s, 1H), 4.08 (brs, 1H), (m, 1H), 3.77 (s, 3H), (m, 1H), (m, 2H), 2.93 (d, 1H), 2.31 (dd, J = 10.4, 2.2 Hz, 1H), 2.12 (s, 3H), (m, 6H), (m, 1H), (m, 1H), 1.30 (d, J = 6.2 Hz, 3H), 1.11 (d, J = 5.9 Hz, 3H), 1.11 (s, 9H), 0.92 (s, 9H),

76 (d, J = 6.3 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.3, 170.2, 155.1, 151.4, 136.3, 136.2, 136.1, 134.2, 134.0, 133.7, 130.0, 129.8, 129.7, 127.9, 127.7, 127.6, 127.6, 117.9, 114.6, 98.9, 97.1, 79.3, 79.1, 77.4, 75.9, 69.6, 69.5, 69.1, 55.7, 35.8, 35.6, 27.4, 27.3, 21.4, 19.7, 19.4, 18.6, 18.4, 18.2; HRMS (ESI) calcd for C 61 H 78 O 13 Si 2 Na [M+Na] , found Trisaccharide 17 To a solution of 16 (1.92 g, 1.78 mmol) in THF (5 ml) was added TBAF (1 M in THF, 9.9 ml, 9.9 mmol) at 0 o C. After stirring for 12 h at rt, the solution was diluted with CH 2 Cl 2, washed with water and brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 2:1) to afford 17 (1.02 g, 96%) as a white foam: [α] 25 D = (c = 1.3, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.95 (d, J = 9.0 Hz, 2H), 6.80 (d, J = 9.1 Hz, 2H), 5.45 (d, J = 3.0 Hz, 1H), 5.34 (d, J = 2.9 Hz, 1H), 5.22 (dd, J = 9.0, 1.6 Hz, 1H), 4.81 (d, J = 8.1 Hz, 1H), 4.71 (d, J = 8.2 Hz, 1H), 4.05 (d, J = 2.9 Hz, 1H), (m, 1H), (m, 1H), 3.75 (s, 3H), (m, 1H), 3.40 (dd, J = 9.0, 3.0 Hz, 1H), 3.25 (ddd, J = 17.2, 9.5, 2.9 Hz, 2H), 2.58 (brs, 1H), 2.40 (brs, 1H), (m, 1H), 2.09 (s, 3H), 2.08 (s, 3H), (m, 3H), (m, 2H), 1.29 (d, J = 6.2 Hz, 3H), 1.21 (d, J = 6.0 Hz, 6H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.5, 170.4, 155.1, 151.4, 117.9, 114.6, 98.9, 98.8, 97.1, 79.3, 79.2, 72.9, 70.1, 69.7, 69.4, 69.2, 68.2, 55.8, 37.8, 36.0, 35.6, 21.5, 21.4, 18.4, 18.1; HRMS (ESI) calcd for C 29 H 42 O 13 Na [M+Na] , found Trisaccharide 18 A solution of 17 (88 mg, mmol) in methanol (3 ml) was refluxed in the presence of dibutyltin oxide (44 mg, mmol) for 4 h. The solvent was then 76

77 removed in vacuo. The residue was dried azeotropically with toluene and then dissolved in DMF (5 ml). Benzyl bromide (0.034 ml, mmol) and CsF (45 mg, mmol) were added, the resulting mixture was stirred at rt for 12 h. The solution was diluted with CH 2 Cl 2, washed with water and brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 2:1) to provide a mixture of acetyl group transfer products as a white foam. To a solution of the above mixture in THF/H 2 O (6 ml, 5:1) at rt was added LiOH (35 mg, 1.47 mmol). The mixture was stirred overnight and was then quenched with addition of the ph = 7.0 buffering solution (8 ml). The mixture was extracted with CH 2 Cl 2. The combined organic layer was washed with brine, dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1:1) to afford 18 (77 mg, 87%) as a white solid: [α] 28 D = (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 5H), 6.96 (d, J = 9.1 Hz, 2H), 6.80 (d, J = 9.1 Hz, 2H), 5.39 (dd, J = 9.1, 2.0 Hz, 1H), 4.91 (d, J = 9.4 Hz, 2H), 4.63 (d, J = 11.4 Hz, 1H), 4.53 (d, J = 11.4 Hz, 1H), 4.32 (s, 1H), (m, 2H), (m, 1H), (m, 2H), 3.75 (s, 3H), 3.32 (dd, J = 8.9, 2.9 Hz, 1H), 3.20 (dd, J = 9.4, 2.8 Hz, 1H), 3.11 (dd, J = 9.4, 2.8 Hz, 1H), 3.06 (s, 1H), 2.96 (s, 1H), 2.43 (d, J = 1.6 Hz, 1H), (m, 1H), (m, 2H), (m, 1H), (m, 2H), 1.27 (d, J = 3.7 Hz, 3H), 1.26 (d, J = 3.6 Hz, 3H), 1.23 (d, J = 6.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 154.9, 151.4, 137.4, 128.8, 128.4, 128.2, 117.9, 114.5, 98.5, 98.4, 96.7, 82.4, 82.3, 80.2, 72.0, 68.8, 68.6, 68.4, 66.5, 66.2, 64.7, 55.8, 36.9, 36.8, 36.5, 18.5, 18.4, 18.3; HRMS (ESI) calcd for C 32 H 44 O 11 Na [M+Na] , found Trisaccharide 19 To a solution of 18 (717 mg, 1.19 mmol) in DMF (10 ml) were added sodium hydride (60% in oil; 381 mg, 9.52 mmol) and methyl iodide (0.37 ml, 5.93 mmol) at 77

78 0 C. The mixture was stirred for 3 h while warming to rt. The solution was quenched with MeOH, diluted with CH 2 Cl 2, washed with brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 4:1) to afford 19 (760 mg, 99%) as a colorless syrup: [α] 28 D = (c = 1.1, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ (m, 5H), 6.96 (d, J = 9.1 Hz, 2H), 6.80 (d, J = 9.1 Hz, 2H), 5.31 (dd, J = 9.3, 1.7 Hz, 1H), (m, 2H), 4.65 (d, J = 11.9 Hz, 1H), 4.52 (d, J = 11.8 Hz, 1H), (m, 1H), (m, 3H), 3.81 (d, J = 2.7 Hz, 1H), (m, 4H), 3.45 (s, 3H), 3.44 (s, 3H), 3.43 (s, 3H), 3.31 (dd, J = 9.4, 2.8 Hz, 1H), 3.23 (dd, J = 9.6, 2.8 Hz, 1H), 3.10 (dd, J = 9.4, 2.7 Hz, 1H), (m, 1H), (m, 2H), 1.83 (ddd, J = 13.4, 9.6, 2.3 Hz, 1H), (m, 2H), 1.27 (d, J = 3.7 Hz, 3H), 1.26 (d, J = 3.6 Hz, 3H), 1.23 (d, J = 6.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 154.9, 151.5, 138.1, 128.5, 128.0, 127.9, 117.7, 114.5, 99.7, 99.7, 96.9, 82.5, 82.1, 80.7, 77.1, 76.8, 74.1, 71.5, 69.0, 69.0, 68.6, 58.1, 58.0, 58.0, 55.7, 35.3, 35.0, 34.9, 18.6, 18.4, 18.3; HRMS (ESI) calcd for C 35 H 50 O 11 Na [M+Na] , found Cymarose trisaccharide 5 To a solution of 19 (315 mg, mmol) in EtOAc/MeOH (12 ml, 2:1) were added Pd(OH) 2 /C (315 mg) and Et 3 N (0.68 ml, 4.87 mmol). The suspension was stirred under hydrogen pressure (1 atm) for 2 days at 50 C and then filtered. The filtrate was concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1:1) to afford 5 (252 mg, 93%) as a white solid: [α] 28 D = (c = 1.5, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.95 (d, J = 9.0 Hz, 2H), 6.79 (d, J = 9.0 Hz, 2H), 5.30 (dd, J = 9.4, 1.7 Hz, 1H), 4.78 (d, J = 8.2 Hz, 1H), 4.68 (d, J = 8.3 Hz, 1H), (m, 1H), (m, 2H), 3.81 (d, J = 2.6 Hz, 1H), 3.75 (s, 3H), 3.61 (d, J = 2.9 Hz, 1H), (m, 1H), 3.45 (s, 3H), 3.44 (s, 3H), 3.42 (s, 3H), 3.31 (dd, J = 9.4, 2.7 Hz, 1H), (m, 2H), (m, 3H), 2.14 (d, J = 12.9 Hz, 1H), (m, 1H), (m, 2H), 1.26 (d, J = 6.2 Hz, 6H), 1.22 (d, J = 6.2 Hz, 78

79 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 154.9, 151.5, 117.8, 114.6, 99.8, 99.6, 96.9, 82.6, 82.2, 77.6, 76.8, 72.6, 70.8, 69.1, 68.6, 58.2, 58.0, 57.3, 55.8, 35.5, 34.9, 33.9, 18.4, 18.4, 18.3; HRMS (ESI) calcd for C 28 H 44 O 11 Na [M+Na] , found Tetrasaccharide 20 To a solution of 4 (270 mg, mmol) and 5 (152 mg, mmol) in toluene (5 ml) was added 5Å MS at rt. After stirring for 30 min at 50 o C, PPh 3 AuNTf 2 (40 mg, mmol) was added to the mixture. The mixture was stirred for 3 h while warming to rt, then Et 3 N was added to quench the reaction. The resulting mixture was filtered and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 2:1) to afford 20 (169 mg, 74%) and its α anomer (42 mg, 19%) as white foams. 20: [α] 30 D = (c = 0.5, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 6.95 (d, J = 9.1 Hz, 2H), 6.80 (d, J = 9.1 Hz, 2H), 5.37 (d, J = 3.0 Hz, 1H), 5.31 (dd, J = 9.3, 1.7 Hz, 1H), 5.08 (dd, J = 9.9, 8.1 Hz, 1H), 4.78 (d, J = 9.6 Hz, 2H), 4.44 (d, J = 8.0 Hz, 1H), 4.20 (d, J = 0.9 Hz, 2H), (m, 1H), (m, 3H), 3.80 (d, J = 2.6 Hz, 1H), 3.76 (s, 3H), (m, 2H), 3.45 (s, 3H), 3.43 (s, 6H), (m, 1H), 3.35 (s, 3H), 3.30 (dd, J = 9.5, 2.8 Hz, 1H), 3.20 (ddd, J = 12.5, 9.7, 2.7 Hz, 2H), (m, 1H), (m, 2H), 2.07 (s, 3H), (m, 1H), (m, 2H), 1.27 (d, J = 6.2 Hz,3H), 1.24 (d, J = 6.2 Hz, 3H), 1.22 (d, J = 6.2 Hz, 3H), 1.17 (d, J = 6.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 169.5, 167.6, 154.9, 151.5, 117.7, 114.6, 102.6, 99.8, 99.7, 96.9, 84.1, 82.6, 82.2, 80.0, 77.0, 76.8, 76.4, 70.7, 70.5, 69.1, 69.1, 68.5, 68.1, 58.3, 58.2, 58.0, 58.0, 55.8, 41.0, 35.5, 35.2, 34.9, 21.0, 18.4, 18.4, 18.2, 16.7; HRMS (ESI) calcd for C 39 H 59 ClO 17 Na [M+Na] , found Tetrasaccharide donor 2 To a solution of 20 (169 mg, mmol) in CH 3 CN/H 2 O (2.5 ml/2.5 ml) was 79

80 added Ag(DPAH) 7 2 (205 mg, mmol) at 0 C. After stirring for 10 min at this temperature, the mixture was filtered. The filtrate was diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/ EtOAc = 1:2) to yield the corresponding hemiacetal (140 mg, 95%) as a colorless syrup. To a solution of the above hemiacetal (84 mg, mmol) in CH 2 Cl 2 (3 ml) were added 2-(cyclopropylethynyl)benzoic acid (43 mg, mmol) S2, DMAP (28 mg, mmol), EDCI (55 mg, mmol), and 4 Å MS (250 mg). The mixture was stirred at rt for 4 h, and then filtered. The filtrate was diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1:1) to afford 2 (102 mg, 99%, β/α = 4.5/1) as a syrup. 2β: [α] 31 D = (c = 0.7, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.91 (d, J = 7.8 Hz, 1H), 7.46 (d, J = 7.6 Hz, 1H), 7.39 (t, J = 7.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 1H), 6.22 (dd, J = 9.3, 1.8 Hz, 1H), 5.36 (d, J = 3.1 Hz, 1H), 5.07 (dd, J = 9.9, 8.1 Hz, 1H), 4.78 (dd, J = 9.4, 2.3 Hz, 2H), 4.44 (d, J = 8.0 Hz, 1H), 4.19 (s, 2H), (m, 1H), (m, 3H), 3.81 (d, J = 2.4 Hz, 1H), (m, 2H), 3.47 (s, 3H), 3.44 (s, 6H), (m, 2H), 3.35 (s, 3H), 3.20 (ddd, J = 15.5, 9.6, 2.6 Hz, 2H), (m, 1H), (m, 2H), 2.07 (s, 3H), (m, 1H), (m, 2H), (m, 1H), 1.27 (d, J = 6.5 Hz, 3H), 1.25 (d, J = 6.5 Hz, 3H), 1.22 (d, J = 6.2 Hz, 3H), 1.18 (d, J = 6.2 Hz, 3H), 0.88 (d, J = 6.2 Hz, 4H); 13 C NMR (100 MHz, CDCl 3 ) δ 169.5, 167.6, 164.5, 134.3, 131.9, 131.4, 130.7, 127.0, 125.1, 102.6, 99.8, 99.7, 92.1, 84.1, 82.6, 81.8, 80.1, 77.4, 76.5, 76.4, 74.7, 70.8, 70.6, 70.2, 69.2, 68.6, 68.2, 58.4, 58.2, 58.1, 57.9, 41.0, 35.7, 35.3, 33.7, 21.0, 18.4, 18.3, 18.2, 16.7, 9.0, 0.8; HRMS (ESI) calcd for C 44 H 61 ClO 17 Na [M+Na] , found Synthesis of FABO disaccharide 25 p-methoxyphenyl 4-O-acetyl-3-O-(tert-butyldimethyl)silyl-2,6-dideoxy-2-iodo-β-D-glucopyranoside (S10) 80

81 To a solution of S9 8,9 (1.32 g, 2.79 mmol) and p-methoxyphenol (624 mg, 5.03 mmol) in CH 2 Cl 2 (10 ml) was added 4 Å MS (2.5 g) at rt. After stirring for 1 h at 72 o C, TMSOTf (0.43 ml, 2.24 mmol) was added to the mixture. After stirring for another 3 h at this temperature, Et 3 N was added to quench the reaction. The resulting mixture was filtered, diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, respectively, dried over Na 2 SO 4, and then concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 40:1) to afford S10 (1.2 g, 80%) as a syrup: [α] 28 D = (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.06 (d, J = 9.0 Hz, 2H), 6.83 (d, J = 9.0 Hz, 2H), 5.00 (d, J = 8.5 Hz, 1H), 4.77 (t, J = 8.8 Hz, 1H), (m, 2H), 3.77 (s, 3H), (m, 1H), 2.11 (s, 3H), 1.23 (d, J = 6.2 Hz, 3H), 0.92 (s, 9H), 0.29 (s, 3H), 0.06 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.0, 155.8, 151.4, 119.1, 114.6, 103.0, 77.1, 76.7, 70.6, 55.8, 36.0, 26.3, 21.7, 18.4, 17.9, -2.8, -3.3; HRMS (ESI) calcd for C 21 H 33 IO 6 SiNa [M+Na] , found p-methoxyphenyl 4-O-acetyl-2,6-dideoxy-2-iodo-β-D-glucopyranoside (11) To a solution of S10 (1.26 g, 2.36 mmol) in MeCN (10 ml) was added 3HF Et 3 N (1.90 ml, 11.8 mmol) at rt. After stirring for 10 h at 65 o C, a saturated NaHCO 3 solution was added slowly to the mixture at rt. The resulting mixture was diluted with EtOAc, washed with saturated NaHCO 3 solution, and was then extracted with EtOAc twice. The combined organic layer was washed with brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 4:1) to afford 11 (930 mg, 94%) as a white solid: [α] 27 D = (c = 1.3, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.07 (d, J = 9.1 Hz, 2H), 6.84 (d, J = 9.1 Hz, 2H), 4.99 (d, J = 9.0 Hz, 1H), 4.72 (t, J = 9.3 Hz, 1H), 4.07 (dd, J = 10.7, 9.1 Hz, 1H), 3.85 (td, J = 10.6, 3.8 Hz, 1H), 3.78 (s, 3H), (m, 1H), 2.79 (brs, 1H), 81

82 2.13 (s, 3H), 1.29 (d, J = 6.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.8, 155.9, 151.3, 119.2, 114.7, 102.6, 76.5, 75.7, 70.5, 55.8, 35.8, 21.0, 17.5; HRMS (ESI) calcd for C 15 H 19 IO 6 Na [M+Na] , found O-Benzoyl-3-O-methyl-6-O-tosyl-D-glucal (S12) To a solution of S11 10 (454 mg, 1.51 mmol) in THF (10 ml) was added TBAF (1 M in THF, 3.8 ml, 3.8 mmol) at 0 o C. After stirring for 3 h at rt, the solution was concentrated. The residue was purified by flash chromatography (CH 2 Cl 2 /MeOH = 30:1) to afford a colorless syrup. The above syrup was dissolved in CH 2 Cl 2 (5 ml) and pyridine (5 ml). TsCl (720 mg, 3.78 mmol) was added to the mixture at 0 o C. After stirring for 22 h at rt, BzCl (0.35 ml, 3.0 mmol) was added at 0 o C. After stirring for 2 h at rt, the mixture was quenched with water at 0 o C. The mixture was diluted with CH 2 Cl 2, washed with saturated NaHCO 3 solution and brine, dried over Na 2 SO 4, and then concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 8:1) to afford S12 (543 mg, 86%) as a colorless syrup: [α] 28 D = (c = 1.1, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.99 (d, J = 7.1 Hz, 2H), 7.77 (d, J = 8.3 Hz, 2H), 7.58 (t, J = 7.4 Hz, 1H), 7.44 (t, J = 7.7 Hz, 2H), 7.29 (d, J = 8.1 Hz, 2H), 6.34 (d, J = 6.3 Hz, 1H), 5.30 (t, J = 4.0 Hz, 1H), (m, 1H), (m, 1H), 4.36 (dd, J = 11.0, 7.6 Hz, 1H), 4.22 (dd, J = 11.0, 3.5 Hz, 1H), (m, 1H), 3.35 (s, 3H), 2.40 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 165.3, 145.0, 144.1, 133.6, 132.9, 129.9, 129.4, 128.6, 128.1, 99.3, 73.5, 71.4, 67.2, 67.2, 56.0, 21.7; HRMS (ESI) calcd for C 21 H 22 SO 7 Na [M+Na] , found O-Benzoyl-6-bromo-6-deoxy-3-O-methyl-D-glucal (21) OTs NaBr, NaHCO 3, BzO MeO O DMF, 70 o C 90% S12 BzO MeO Br O 21 To a solution of S12 (596 mg, 1.42 mmol) in DMF (10 ml) were added NaBr (733 82

83 mg, 7.12mmol) and NaHCO 3 (358 mg, 4.26 mmol) at rt. After stirring for 6 h at 70 o C, the mixture was cooled to rt and concentrated. Diluted with EtOAc, this mixture was washed with water and brine, dried over Na 2 SO 4, and then concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 30:1) to afford 21 (420 mg, 90%) as a colorless syrup: [α] 27 D = 2.9 (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 8.05 (d, J = 7.4 Hz, 2H), 7.59 (t, J = 7.4 Hz, 1H), 7.45 (t, J = 7.7 Hz, 2H), 6.51 (d, J = 6.3 Hz, 1H), 5.56 (t, J = 4.3 Hz, 1H), 5.00 (dd, J = 5.9, 4.4 Hz, 1H), 4.45 (dd, J = 12.0, 5.9 Hz, 1H), 3.89 (t, J = 3.8 Hz, 1H), (m, 2H), 3.44 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 165.4, 144.3, 133.6, 130.0, 129.5, 128.6, 99.6, 75.4, 72.1, 68.4, 56.2, 29.7; HRMS (ESI) calcd for C 14 H 15 BrO 4 Na [M+Na] , found O-Benzoyl-6-bromo-2,6-dideoxy-3-O-methyl-2-(phenyl)seleno-D-mannopyrano side (22) To a stirring solution of 21 (1.77 g, 5.41 mmol) in CH 3 CN (30 ml) was added a solution of PhSeCl (1.14 g, 5.95 mmol) in CH 3 CN (10 ml) dropwise at 40 o C. The reaction was monitored by TLC until no glycal remained, then saturated NaHCO 3 solution was added. The mixture was extracted with CH 2 Cl 2. The combined organic layer was washed with brine, dried over Na 2 SO 4, and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 8:1) to afford 22 (2.1 g, 78%) as a white solid. 22α: [α] 27 D = +6.9 (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 8.06 (d, J = 8.4 Hz, 2H), (m, 2H), 7.59 (t, J = 7.4 Hz, 1H), 7.46 (t, J = 7.7 Hz, 2H), (m, 3H), 5.55 (t, J = 3.0 Hz, 1H), 5.40 (t, J = 8.5 Hz, 1H), (m, 1H), 4.07 (dd, J = 8.1, 4.4 Hz, 1H), 3.83 (dd, J = 4.4, 2.7 Hz, 1H), (m, 2H), 3.31 (s, 3H), 3.16 (d, J = 3.7 Hz, 1H); 13 C NMR (100 MHz, CDCl 3 ) δ 165.7, 135.3, 133.6, 130.0, 129.6, 129.3, 129.1, 128.6, 128.1, 95.1, 78.2, 71.6, 71.4, 57.8, 48.3, 32.2; MS (ESI) [M+Na]

84 Phenylthiomethyl 4-O-benzoyl-6-bromo-2,6-dideoxy-3-O-methyl-2-(phenyl)seleno-α-D-mannopyran oside (10) To a solution of 22 (671 mg, 1.34 mmol) in THF (5 ml) was added dimethylaminosulfur trifluoride (DAST) (0.41 ml, 3.35 mmol) at 30 o C. After stirring for 3 h at rt, a saturated NaHCO 3 solution was added slowly to the mixture. The resulting mixture was extracted with CH 2 Cl 2. The combined organic layer was washed with brine, dried over Na 2 SO 4, and concentrated. The crude glycosyl fluoride was azeotroped with toluene (3 x 5 ml). After drying under high vacuum for at least 2 h, the above product was dissolved in CH 2 Cl 2 (8 ml) and 4 Å MS (1.5 g) was added. The reaction mixture was stirred at 40 o C for 30 min before PhSCH 2 OH 11 (366 mg, 2.68 mmol) and SnCl 2 (381 mg, 2.01 mmol) were added. The reaction mixture was allowed to warm to rt and stirred for 1 h. The reaction mixture was quenched with Et 3 N (1 ml) and filtered. The solution was diluted with EtOAc, and washed with water. The water layer was extracted with EtOAc twice. The combined organic layer was washed with saturated NaHCO 3 solution and brine, dried over Na 2 SO 4, and then concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 60:1) to afford 10 (480 mg, 58%) as a syrup: [α] 28 D = (c = 1.1, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 8.04 (d, J = 7.2 Hz, 2H), (m, 3H), 7.46 (t, J = 7.7 Hz, 2H), 7.38 (dd, J = 7.9, 1.4 Hz, 2H), (m, 6H), 5.56 (d, J = 1.8 Hz, 1H), 5.33 (t, J = 8.9 Hz, 1H), 5.13 (d, J = 11.7 Hz, 1H), 5.06 (d, J = 11.8 Hz, 1H), (m, 1H), 3.96 (dd, J = 8.4, 4.7 Hz, 1H), 3.77 (dd, J = 4.5, 2.1 Hz, 1H), (m, 2H), 3.29 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 165.6, 135.2, 135.0, 133.6, 130.8, 130.0, 129.6, 129.3, 129.1, 129.0, 128.6, 128.1, 127.3, 97.1, 78.5, 71.8, 71.7, 71.7, 57.7, 47.4, 31.9; MS (ESI) [M+Na]

85 Acetal glycoside 24 To a solution of 11 (620 mg, 1.47 mmol) in CH 2 Cl 2 (5 ml) was added acid washed 3 Å MS at rt. After stirring for 30 min at rt, NIS (142 mg, mmol) and TfOH (2.6 μl, mmol) were added to the mixture at 30 ºC, followed by addition of a solution of 10 (183 mg, mmol) in CH 2 Cl 2 (3 ml) via a syringe pump. The reaction mixture was stirred at 30 C for 1 h. When TLC showed the donor had been consumed, saturated aqueous NaHCO 3 was added at 0 ºC. The mixture was filtered. The filtrate was washed with a solution of NaS 2 O 3 and brine, respectively, and was then dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (petroleum ether/ CH 2 Cl 2 /EtOAc = 8:1:1) to afford 24 (206 mg, 75%) and 23 (53 mg, 20%) as syrups. 24: [α] 28 D = (c = 1.2, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 8.07 (d, J = 7.4 Hz, 2H), 7.69 (d, J = 7.7 Hz, 2H), 7.60 (t, J = 7.4 Hz, 1H), 7.46 (t, J = 7.7 Hz, 2H), (m, 3H), 7.05 (d, J = 9.0 Hz, 2H), 6.84 (d, J = 9.0 Hz, 2H), 5.70 (s, 1H), 5.37 (t, J = 9.4 Hz, 1H), 5.08 (d, J = 6.5 Hz, 1H), 4.96 (d, J = 8.9 Hz, 1H), 4.91 (d, J = 6.5 Hz, 1H), 4.84 (t, J = 9.2 Hz, 1H), (m, 2H), (m, 2H), (m, 1H), 3.78 (s, 3H), (m, 1H), (m, 2H), 3.28 (s, 3H), 2.15 (s, 3H), 1.24 (d, J = 6.3 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 169.5, 165.7, 156.0, 151.2, 136.1, 133.6, 130.0, 129.6, 129.3, 128.8, 128.6, 128.4, 119.2, 114.7, 102.9, 97.8, 91.3, 81.1, 77.9, 76.2, 72.2, 71.8, 70.5, 57.8, 55.8, 48.1, 32.0, 31.7, 21.1, 17.5; MS (ESI) [M+Na] : [α] 28 D = (c = 0.9, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 8.08 (d, J = 7.5 Hz, 2H), (m, 3H), 7.47 (t, J = 7.7 Hz, 2H), (m, 3H), 7.08 (d, J = 9.0 Hz, 2H), 6.85 (d, J = 9.0 Hz, 2H), 5.62 (t, J = 8.0 Hz, 1H), 5.59 (d, J = 2.7 Hz, 1H), 5.03 (d, J = 8.9 Hz, 1H), 4.90 (t, J = 9.1 Hz, 1H), (m, 1H), (m, 1H), (m, 2H), 3.78 (s, 3H), (m, 2H), (m, 2H), 3.26 (s, 3H), 1.98 (s, 3H), 1.23 (d, J = 6.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 170.0, 165.5, 156.1, 151.2, 134.5, 133.5, 130.1, 129.8, 129.7, 129.2, 128.6, 127.9, 119.3, 114.7, 103.0, 101.8, 80.2, 78.7, 76.6, 71.6, 85

86 70.6, 70.4, 58.2, 55.8, 48.4, 32.8, 31.7, 21.1, 17.4; MS (ESI) [M+Na] Disaccharide 9 Br BzO MeO SePh O AcO O O 24 O I OMP MeONa, MeOH, CH 2 Cl 2,rt 93% Br HO MeO SePh O HO O O 9 O I OMP To a solution of 24 (236 mg, mmol) in CH 2 Cl 2 /MeOH (3 ml/3 ml) was added NaOMe (28 mg, mmol) at rt. After stirring for 9 h at rt, the mixture was filtered through silica gel. The filtrate was evaporated in vacuo to give a residue, which was purified by flash chromatography (petroleum ether/ EtOAc = 2:1) to afford 9 (185 mg, 93%) as a colorless syrup: [α] 30 D = 7.6 (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 7.65 (dd, J = 7.4, 1.8 Hz, 2H), (m, 3H), 7.05 (d, J = 9.0 Hz, 2H), 6.84 (d, J = 9.0 Hz, 2H), 5.49 (s, 1H), 5.13 (d, J = 6.5 Hz, 1H), 4.97 (d, J = 6.5 Hz, 1H), 4.92 (d, J = 9.1 Hz, 1H), 3.95 (dd, J = 10.6, 9.2 Hz, 1H), (m, 3H), 3.78 (s, 3H), (m, 2H), 3.61 (dd, J = 11.0, 6.2 Hz, 1H), 3.53 (dd, J = 10.6, 8.4 Hz, 1H), (m, 1H), 3.33 (s, 3H), (m, 1H), 3.15 (d, J = 2.5 Hz, 1H), 2.71 (s, 1H), 1.37 (d, J = 6.0 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 155.8, 151.4, 135.7, 129.5, 128.7, 128.5, 119.0, 114.7, 102.7, 98.9, 93.8, 86.5, 79.8, 76.2, 72.7, 72.1, 69.9, 56.8, 55.8, 47.1, 33.0, 32.3, 17.7; MS (ESI) [M+Na] FABO disaccharide 25 To a solution of 9 (187 mg, mmol) in MeOH/CH 2 Cl 2 /H 2 O (3 ml/2 ml/1 ml) were added NaIO 4 (507 mg, 2.37 mmol) and NaHCO 3 (159 mg, 1.90 mmol) at rt. After stirring for 3 h at rt, the mixture was diluted with CH 2 Cl 2, washed with saturated NH 4 Cl solution and brine, and was then dried over Na 2 SO 4 and concentrated. The resultant selenoxide was azeotroped with toluene (3 x 5 ml) and dried under high vacuum for at least 2 h to afford a white solid (190 mg, 99%). The above selenoxide (125 mg, mmol) was dissolved in toluene (6 ml). Diisopropylamine (3 ml) and vinyl acetate (6 ml) were added, and the reaction was conducted under microwave at 86

87 145 ºC for 20 min. The mixture was cooled to rt and concentrated. The residue was purified by flash chromatography (petroleum ether/etoac = 1.5:1) to afford 25 (83 mg, 85%) as a colorless syrup: [α] 25 D = (c = 0.8, CHCl 3 ); 1 H NMR (500 MHz, CDCl 3 ) δ 7.08 (d, J = 9.1 Hz, 2H), 6.84 (d, J = 9.1 Hz, 2H), 5.12 (d, J = 8.0 Hz, 1H), 5.01 (d, J = 9.1 Hz, 1H), 4.90 (d, J = 8.0 Hz, 1H), 4.04 (dd, J = 10.9, 9.1 Hz, 1H), 3.99 (t, J = 8.9 Hz, 1H), (m, 2H), 3.78 (s, 3H), (m, 1H), 3.52 (dd, J = 11.2, 7.9 Hz, 2H), (m, 1H), 3.41 (s, 3H), (m, 1H), 2.66 (brs, 1H), 2.53 (dd, J = 12.7, 4.6 Hz, 1H), 1.59 (t, J = 12.1 Hz, 1H), 1.46 (d, J = 6.2 Hz, 3H); 13 C NMR (125 MHz, CDCl 3 ) δ 156.0, 151.4, 119.2, 114.7, 114.7, 103.4, 88.9, 84.2, 78.9, 76.7, 74.0, 72.4, 70.7, 56.9, 55.8, 37.1, 32.4, 29.7, 19.1; HRMS (ESI) calcd for C 21 H 28 BrIO 9 Na [M+Na] , found Preparation of pregnane diol 7 and its derivative S14 for X-ray diffraction analysis To a stirred solution of AD-mix-β 12 (18.6 g) in t-buoh/h 2 O (80 ml/80 ml) were added methanesulfonamide CH 3 SO 2 NH 2 (3.44 g, 36.2 mmol) and a solution of S13 13 (5.0 g, 12.1 mmol) in CH 2 Cl 2 (40 ml) at 0 C. The solution was kept at 0 C for 2 days and then kept at rt for another 3 days. Solid Na 2 SO 3 was added, and the solution was stirred for 1 h. The solution was extracted with EtOAc. The resulting organic layer was washed with 2 N KOH and brine, respectively, and was then dried over Na 2 SO 4 and concentrated. The residue was purified by flash chromatography (CH 2 Cl 2 / EtOAc = 10:1) to afford 7 (4.57 g, 95% based on 85% conversion, d.r. = 96:4) as a white solid and recovered S13 (0.53 g, 10%). 7: [α] 27 D = 58.8 (c = 1.0, CHCl 3 ); 1 H NMR (400 MHz, CDCl 3 ) δ 5.31 (brs, 1H), (m, 1H), (m, 1H), (m, 1H), (m, 1H), (m, 3H), (m, 3H), (m, 4H), (m, 7H), 1.18 (d, J = 6.3 Hz, 3H), (m, 1H), 0.99 (s, 3H), (m, 1H), 0.88 (s, 9H), 0.73 (s, 3H), 0.05 (s, 6H); 13 C NMR 87