Supporting Information Pentavalent Bismuth as a Universal Promoter for S-Containing Glycosyl Donors with a Thiol Additive Daniel E. K.

Transcription

1 Supporting Information Pentavalent Bismuth as a Universal Promoter for S-Containing Glycosyl Donors with a Thiol Additive Daniel E. K. Kabotso and Nicola L. B. Pohl * Department of Chemistry, Indiana University, 120A Simon Hall, 212 South Hawthorne Drive, Bloomington, Indiana 47405, United States 1

2 Table of Contents General Information... 3 Material... 3 General Experimental Procedure... 4 General glycosylation protocol using Ph3Bi(OTf) General protocol for fluorous solid phase extraction... 4 Experimental procedures... 4 References NMR Spectra ( 1 H and 13 C NMR, HSQC, 1 H- 13 C HSQC)

3 General Information Every reaction was carried out under argon atmosphere in an oven dried round bottomed (RB) flask unless otherwise stated. Chemicals and reagents used were purchased from Sigma-Aldrich, Alfa Aesar, and Acros Chemicals and used without further purification. Molecular Sieves, Activated, Type 4A, 8 to 12 Mesh, BAKER ANALYZED Reagent, J.T.Baker (purchased from Capitol Scientific, Inc Rutland Dr. Austin, TX USA) were dried at 300 o C for 12 h in Thermolyne Industrial Benchtop Muffle Furnace (Thermo Scientific) before used. All solvents as media for reactions were dried over molecular sieves for 12 h and then used without purification. The progress of reactions was monitored via TLC analysis, performed on Hard Layer TLC plates (silica gel w/uv254, glass backed 250µm, 20 x 20 cm, purchased from Sorbent Technologies, 5955 Peachtree Corners East Suite A, Norcross GA USA). Detection of compounds on TLC plates was accomplished by two means; 1) exposure of the plates to UV light (2536Å) and 2) dipping of the plate in p-anisaldehyde dip (made of ethanol:panisaldehyde:sulfuric acid = 18:1:1). Purification of compounds was achieved by performing fluorous solid phase extraction (by using 5 g size prepacked fluorous column purchased from Fluorous Technologies c/o Boron Specialties LLC, 2301 Duss Ave, Ste 35, Ambridge, PA USA) and medium pressure liquid chromatography using Teledyne ISCO (purchased from Teledyne Isco 4700 Superior Street Lincoln, NE U.S.A.). Proton ( 1 H) NMR and carbon ( 13 C) NMR, spectra were recorded on a 500 MHz instrument using the residual signals from CDCl 3, δ7.26 ppm, and 77.0 ppm as an internal reference for 1 H and 13 C chemical shifts, respectively. Chemical shifts in 1 H-NMR spectra are reported in ppm (δ) with reference to the signal of Me4Si, which was adjusted to δ 0.00 ppm unless otherwise noted. Multiplicities of signals were represented as follow: s = singlet, d =doublet, t = triplet, dd = double of doublet, td = triple doublet, m = multiplets. Integration and coupling constants are given in Hertz (Hz). Heteronuclear single quantum coherence spectroscopy (HSQC) was used to determine the stereochemistry of the products where necessary according published procedure. 1,2 ESI-HRMS mass spectrometry was obtained using Agilent 6540 QTOF instrument with Bruker AVANCE III 500 spectrometer. Material The pentavalent bismuth promoter was made according to literature procedure 1,2 from triphenyl bismuth, triflic acid (TfOH) and (diacetoxyiodo)benzene (BAIB) purchased from Alfa Aesar, Radcliff Rd, Tewksbury, MA 01876, United States. 3

4 The donors such as 2 4, 20, 13 23, 4 24, 5 25, 6,7 26, 8 28, 9,10 29, 9,10 31, 3 44, 11,12 and 45 3 were made according to literature procedures. In addition, acceptors and intermediates or starting substrates, such as 3, 14 21, 15 35, , 19 37, 6,7 and 58 20,21 were constructed based on standard literature protocols. Acceptor 27 was purchased from Boron specialties, LLC, 2301 Dues Avenue, Box 35 Ambridge, PA USA. General Experimental Procedure General glycosylation protocol using Ph3Bi(OTf)2 The donor (2.0 equiv) and the promoter, 1 (2.0 equiv) were taken into a 25-mL RB flask, which was charged with a magnetic stir bar. The flask was purged and filled with argon. To this were added a purged solution of the acceptor (1.0 equiv) and PrSH (2.0 equiv) in anhydrous CH2Cl2 to give a final reaction concentration of mm with respect to the donor. After TLC analysis has shown complete consumption of the acceptor (40 min- 12 h), Et3N (0.1 ml) was added to quench the reaction. The reaction mixture was filtered over a pad of celite, concentrated under reduced pressure and the crude product subjected to fluorous solid phase extraction (FSPE) followed by medium pressure liquid chromatographic (MPLC) purification using Teledyne ISCO to afford the product. General protocol for fluorous solid phase extraction The fluorous column was washed with DMF to remove any silica gel that was not derivatised with fluorine atoms. The column was equilibrated with 80% MeOH/H2O. The crude material was dissolved in a minimum amount of DMF and then loaded onto the fluorous column and allowed to interact with the column under gravity. The non-fluorous component of the crude material was eluted with 80% MeOH/H2O (fluorophobic solvent system) while the fluorous compound was eluted with pure acetone as the fluorophilic solvent. Experimental procedures Glycosylation of donor 2 with acceptor 3 to give product 4 cis-4-(1h,1h,2h,2h,3h,3h-perfluoroundecyloxy)-2-butenyl, 2,3,4,6-tetra-O-acetyl-β-Dgalactopyranoside (4): Phenyl 2,3,4,6-tetra-O-1-thio-β-galactopyranoside, 2 4 (161 mg, 365 µmol) was coupled to cis-4-(1h,1h,2h,2h,3h,3h-perfluoroundecyloxy)-2-buten-1-ol, 3 14,22 4

5 (100.0 mg, 182 µmol) according to the general protocol for glycosylation using Ph3Bi(OTf)2 to afford the targeted crude material, which was purified via FSPE and medium pressure liquid chromatography (MPLC) using ISCO (0-25% EtOAc in hexane) to give the pure product 4 (120 mg, 75%). 1 H NMR (500 MHz, CDCl3) δ(ppm) (m, 2H), 5.37 (dd, J = 3.4, 1.2 Hz, 1H), 5.19 (dt, J = 9.5, 7.2 Hz, 1H), 5.00 (dd, J = 10.4, 3.5 Hz, 1H), 4.49 (d, J = 7.9 Hz, 1H, H-1), 4.34 (dd, J = 5.8, 1.3 Hz, 1H), (m, 4H), (m, 2H), (m, 1H), 3.47 (t, J = 6.0 Hz, 2H), (m, 2H), 2.13 (s, 3H), 2.03 (d, J = 3.4 Hz, 6H), 1.96 (s, 3H), 1.87 (dt, J = 10.1, 6.0 Hz, 2H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , (C- 1), 70.91, 70.70, 68.77, 68.75, 66.99, 66.48, 64.71, 61.21, 28.12, 27.94, 27.77, 20.85, 20.82, 20.79, 20.68, 20.59, 20.54, HRMS-ESI-TOF (m/z): Calcd. for [C29H31F17O11+Na] , Found Glycosylation of donor 23 with acceptor 21 to give product 5 2-(4-((4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoroundecyl)oxy)phenoxy)ethyl 2,3- di-o-benzyl-4,6-di-tert-butylsilyl-α-d-galactopyranoside (5): Treatment of donor 23 6 (199 mg, 335 µmol) with the acceptor (100.0 mg, 168 µmol), according to the general glycosylation protocol using the pentavalent bismuth gave the desired product 5 (157 mg, 85%) after purification via medium pressure liquid chromatography (MPLC) using ISCO (0-10% EtOAc in hexane). 1 H NMR (500 MHz, CDCl3) δ(ppm) (m, 2H), (m, 4H), (m, 4H), 6.82 (s, 4H), 4.90 (d, J = 11.9 Hz, 1H), 4.85 (d, J = 3.7 Hz, 1H, H-1), 4.75 (s, 2H), 4.70 (d, J = 11.9 Hz, 1H), 4.52 (d, J = 3.3 Hz, 1H), (m, 4H), (m, 3H), 3.94 (ddd, J = 11.5, 6.5, 3.6 Hz, 1H), (m, 2H), 3.74 (d, J = 1.9 Hz, 1H), (m, 2H), (m, 2H), 1.08 (s, 9H), 1.02 (s, 9H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , , (C-1), 77.69, 74.27, 73.62, 71.17, 71.08, 67.69, 67.29, 67.23, 66.97, 66.53, 27.98, 27.64, 27.31, 23.42,

6 HRMS-ESI-TOF (m/z): Calcd. For [C47H53NO8Si+Na] , Found Glycosylation of donor 24 5 with acceptor to give product 6 Condensation of 24 5 (145 mg, 326 µmol) with (100.0 mg, 163 µmol), mediated by pentavalent bismuth (240.0 mg, 326 µmol) according to the general glycosylation protocol followed by purification via MPLC using ISCO (0-25% EtOAc in Hexane) gave the titled product 6 (114 mg, 74%). 1 H NMR (500 MHz, CDCl3) δ(ppm) 7.19 (s, 1H), 6.75 (d, J = 0.8 Hz, 4H), 5.33 (dd, J = 3.4, 1.2 Hz, 1H), 5.17 (dd, J = 10.5, 8.0 Hz, 1H), 4.97 (dd, J = 10.5, 3.4 Hz, 1H), 4.57 (d, J = 8.0 Hz, 1H, H-1), (m, 5H), 3.92 (t, J = 5.9 Hz, 2H), 3.86 (td, J = 6.8, 1.1 Hz, 2H), 2.24 (tt, J = 18.5, 8.0 Hz, 2H), 2.08 (s, 3H), (m, 2H), 1.97 (s, 3H), 1.91 (s, 3H), 1.87 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , (C-1), 70.90, 70.75, 68.76, 68.36, 67.96, 67.04, 66.97, 61.27, 27.96, 20.68, 20.65, HRMS-ESI-TOF (m/z): Calcd. for [C33H37F17O12+NH4] , Found Glycosylation of donor 25 6,7 with acceptor 3 14,22 to give product 7 Coupling of the donor 25 5,6 (208 mg, 365 µmol) to the acceptor 3 13,21 (100.0 mg, 182 µmol) in the presence of the pentavalent bismuth (269 mg, 365 µmol) led to the desired product 7 (128 mg, 71%) after purification of the crude product via FSPE and MPLC using ISCO (0-40% EtOAc in Hexane). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 2H), (m, 1H), 5.23 (s, 1H), 5.02 (s, 1H), (m, 3H), 4.39 (dd, J = 12.8, 5.6 Hz, 1H), 4.25 (dd, J = 12.8, 6.9 Hz, 1H), (m, 2H), (m, 2H), 3.90 (t, J = 6.8 Hz, 1H), 3.79 (d, J = 9.0 Hz, 1H), 3.49 (t, J = 6.0 Hz, 2H), 2.17 (s, 2H), 2.15 (s, 3H), 2.05 (s, 3H), 2.00 (s, 3H), 1.88 (dd, J = 10.2, 5.8 Hz, 2H). 6

7 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , 99.89, 70.74, 68.86, 66.66, 66.46, 64.86, 61.29, 52.95, 29.70, 27.98, 27.81, 20.86, 20.66, 20.62, HRMS-ESI-TOF (m/z): Calcd. for [(C41H52NCl3F17O15+NH4] , Found Glycosylation of donor 26 8 with acceptor 27 to give product 8 In the presence of the pentavalent bismuth (266 mg, 361 µmol), donor 26 8 (180.0 mg, 361 µmol) was successfully coupled to the acceptor 27 (100.0 mg, 180 µmol) to give the product 8 (107 mg, 67%) according the general pentavalent bismuth-mediated glycosylation protocol followed by medium pressure liquid chromatographic purification on ISCO using 0-30% EtOAc in Hexane 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 2H), 7.13 (d, J = 8.1 Hz, 2H), (m, 3H), 4.80 (d, J = 12.2 Hz, 1H), 4.53 (d, J = 12.2 Hz, 1H), 4.47 (d, J = 7.9 Hz, 1H, H-1), 4.21 (dd, J = 12.3, 4.7 Hz, 1H), 4.10 (dd, J = 12.3, 2.5 Hz, 1H), 3.61 (ddd, J = 9.8, 4.7, 2.5 Hz, 1H), (m, 2H), 2.30 (tt, J = 17.7, 8.4 Hz, 2H), 2.03 (s, 3H), 1.94 (d, J = 2.9 Hz, 6H), 1.93 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , (C-1), 72.81, 71.87, 71.28, 70.36, 68.43, 61.94, 33.10, 32.92, 32.75, 26.21, 26.17, 26.14, 20.73, 20.63, 20.59, HRMS-ESI-TOF (m/z): Calcd. for [C31H29F17O10+Na] , Found Glycosylation of donor 28 9,10 with acceptor 27 to give product 8 According to the general pentavalent bismuth-mediated glycosylation protocol for thioglycosides, the donor 28 9,10 (162 mg, 361 µmol) and acceptor 27 (100.0 mg, 180 µmol) were successfully condensed in the presence of the pentavalent bismuth promoter (266 mg, 361 µmol) to give the product 8 (104 mg, 65%) after medium pressure liquid chromatographic purification using ISCO (0-25% EtOAc in Hexane). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 2H), 7.13 (d, J = 8.1 Hz, 2H), (m, 3H), 4.80 (d, J = 12.2 Hz, 1H), 4.53 (d, J = 12.2 Hz, 1H), 4.47 (d, J = 7.9 Hz, 1H, H-1),

8 (dd, J = 12.3, 4.7 Hz, 1H), 4.10 (dd, J = 12.3, 2.5 Hz, 1H), 3.61 (ddd, J = 9.8, 4.7, 2.5 Hz, 1H), (m, 2H), 2.30 (tt, J = 17.7, 8.4 Hz, 2H), 2.03 (s, 3H), 1.94 (d, J = 2.9 Hz, 6H), 1.93 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , (C-1), 72.81, 71.87, 71.28, 70.36, 68.43, 61.94, 33.10, 32.92, 32.75, 26.21, 26.17, 26.14, 20.73, 20.63, 20.59, HRMS-ESI-TOF (m/z): Calcd. for [C31H29F17O10+Na] , Found Glycosylation of donor 29 9,10 with acceptor 3 14,22 to give product 9 By coupling donor 29 9,10 (176 mg, 365 µmol) to the acceptor 3 14,22 (100 mg, 182 µmol) with the aid of pentavalent bismuth promoter (269 mg, 365 µmol) according to the general protocol for bismuth mediated glycosylation and purification via MPLC using ISCO (0-25% EtOAc in Hexane), the product 9 (98.2 mg 61%) was obtained 1 H NMR (500 MHz, CDCl3) δ(ppm) (m, 2H), (m, 1H), 5.08 (td, J = 9.7, 4.4 Hz, 1H), 4.99 (dd, J = 9.6, 8.0 Hz, 1H), 4.52 (dd, J = 15.0, 8.0 Hz, 1H), 4.35 (ddd, J = 12.7, 5.8, 1.3 Hz, 1H), (m, 3H), 4.14 (dt, J = 12.5, 3.3 Hz, 1H), (m, 3H), 3.67 (ddd, J = 9.9, 4.7, 2.5 Hz, 1H), 3.49 (dt, J = 12.3, 6.0 Hz, 3H), (m, 2H), 2.07 (d, J = 3.0 Hz, 3H), 2.03 (d, J = 2.0 Hz, 3H), 2.01 (d, J = 2.1 Hz, 3H), 1.99 (d, J = 1.6 Hz, 3H), (m, 2H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , 99.54, 72.83, 71.85, 71.22, 68.86, 68.79, 68.39, 66.49, 66.47, 64.72, 61.91, 58.74, 29.68, 27.96, 20.64, 20.61, 20.56, HRMS-ESI-TOF (m/z): Calcd. for [C29H31F17O11+Na] , Found Scheme S1 Synthesis of acceptor 30 Reagents and Reaction Conditions: a) (i) Na, MeOH; (ii) TrCl, Et3N, DMAP, DMF, 25 o C, 24 h (iii). BnBr, NaH, DMF, 0-25 o C; d. 1% TFA/CH2Cl2, Et3SiH, 23 o C, 30 min, 73% 8

9 4-(1H,1H,2H,2H-Perfluorodecyl)benzyl 234-tri-O-benzyl-β-Dglucopyranoside (30): The dried tetra-ol (0.821 g, 1.15 mmol) resulting from deacetylation of 8 was dissolved in anhydrous N, N- Dimethylformamide (DMF) under argon. To this solution of the tetra-ol was added Et3N (0.578 g, 5.73 mmol, 5 equiv), TrCl (0.399 g, 1.43 mmol, 1.25 equiv), and a catalytic amount of DMAP. The resulting reaction mixture was stirred at 25 o C until TLC analysis confirmed transformation of the starting material into tritylated product (24 h). NaH (0.123 g, 5.16 mmol, 4.5 equiv) and BnBr (0.882 g, 5.16 mmol, 4.5 equiv) were added to the reaction mixture at 0 o C and then gradually taken to room temperature. The reaction was stirred at the 23 o C temperature until TLC indicated completion of the reaction (12 h). The reaction mixture was concentrated under reduced pressure to give the crude material, which washed with saturated solution of ammonium chloride (2x 30 ml). The organic layer was dried (Na2SO4), filtered and concentrated under reduced pressure to give the crude material, which dissolved in 1% TFA in CH2Cl2 and Et3SiH (1.10 ml, 6.88 mmol, 6.0 equiv). The reaction mixture was stirred at 23 o C temperature until TLC analysis confirmed complete removal of the trityl group (30 min). Toluene was added and the reaction mixture evaporated. After co-evaporation for three more times with toluene, the resulting crude material was purified via MPLC using ISCO (0-40% EtOAc in Hexane) to give the titled product 30 (0.830 g, 73%). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 13H), 7.18 (d, J = 7.9 Hz, 2H), (m, 4H), (m, 3H), (m, 1H), (m, 1H), (m, 2H), (m, 1H), 3.49 (dd, J = 9.1, 7.8 Hz, 1H), 3.37 (ddd, J = 9.7, 4.7, 2.8 Hz, 1H), (m, 2H), 2.37 (tq, J = 17.7, 8.2 Hz, 2H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , , 84.56, 82.36, 77.58, 75.73, 75.09, 74.98, 71.28, 62.10, 32.94, 26.21, 26.18, HRMS-ESI-TOF (m/z): Calcd. for [C44H39F17O6+Na] , Found

10 Glycosylation of donor 23 6 with acceptor 30 to give product 10 The donor 23 6 (91.3 mg, 203 µmol), and acceptor 30 (100.0 mg, 101 µmol), in the presence of pentavalent bismuth promoter (150 mg, 203 µmol) and according to the general bismuth-meditated glycosylation protocol for thioglycosides, the product 10 (106 mg, 79%) was obtained after purification via MPLC using ISCO (0-20% EtOAc in Hexane). 1 H NMR (500 MHz, CDCl3) δ(ppm) 7.41 (t, J = 6.4 Hz, 4H), 7.29 (qd, J = 9.3, 7.5, 4.0 Hz, 20H), 7.23 (dq, J = 7.4, 4.2 Hz, 4H), 7.15 (d, J = 7.8 Hz, 2H), 4.97 (d, J = 3.5 Hz, 1H), (m, 5H), (m, 4H), 4.67 (d, J = 11.0 Hz, 1H), 4.59 (dd, J = 22.1, 11.5 Hz, 2H), (m, 2H), (m, 3H), (m, 3H), (m, 3H), 3.53 (t, J = 7.3 Hz, 1H), 3.44 (t, J = 8.2 Hz, 1H), (m, 2H), 2.37 (tt, J = 18.0, 8.1 Hz, 2H), 1.07 (s, 9H), 1.01 (s, 9H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , , , , , , , , , , 97.27, 83.66, 81.29, 75.98, 75.73, 74.56, 73.93, 73.84, 73.63, 73.45, 71.99, 70.34, 69.95, 69.55, 66.22, 66.17, 65.58, 26.63, 26.28, 22.39, 19.61, HRMS-ESI-TOF (m/z): Calcd. for (C72H77F17O11Si+Na) , Found Glycosylation of donor 24 5 with acceptor 30 to give product 11 Subjection of the donor 24 5 (92.1 mg, 203 µmol) and the coupling partner 30 (100.0 mg, 101 µmol) in the pentavalent bismuth (150 mg, 203 µmol) to the general glycosylation protocol for thioglycosides furnished product 11 (99 mg, 74%) after purification via MPLC using ISCO (0-25% EtOAc in Hexane). 1 H NMR (500 MHz, CDCl3) δ(ppm) (m, 18H), 7.11 (d, J = 8.0 Hz, 2H), 5.30 (dd, J = 3.5, 1.2 Hz, 1H), 5.20 (dd, J = 10.4, 8.0 Hz, 1H), (m, 4H), 4.77 (d, J = 11.0 Hz, 1H), 10

11 4.67 (dd, J = 21.6, 10.9 Hz, 2H), 4.59 (d, J = 12.0 Hz, 1H), (m, 2H), 4.40 (d, J = 7.8 Hz, 1H), (m, 3H), 3.77 (td, J = 6.7, 1.2 Hz, 1H), (m, 2H), (m, 2H), 3.33 (dd, J = 9.9, 8.8 Hz, 1H), (m, 2H), 2.30 (tt, J = 18.1, 8.5 Hz, 2H), 2.08 (s, 3H), 1.94 (s, 3H), 1.91 (d, J = 1.8 Hz, 6H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , , , , , , , , , 84.64, 82.21, 78.00, 73, 74.92, 74.89, 74.78, 71.02, 70.68, 70.60, 68.87, 68.43, 67.05, 61.26, 32.93, 29.70, 26.17, 20.83, 20.67, 20.64, HRMS-ESI-TOF (m/z): Calcd. for [C58H57F17O15+Na] , Found Scheme S2: Synthesis of acceptor 34 Reagents and conditions: a) Ph3Bi(OTf)2 (1.0 equiv), PrSH (1.0 equiv), alkene-tag (0.5 equiv), CH2Cl2, 1 h, 87%; b) (i) Na (0.2 equiv), MeOH, 4 h, 94%, (ii) PhCH(OMe) (2.0 equiv), PTSA (0.2 equiv), 77%; c) NaH (3.0 equiv), BnBr (3.0 equiv), CH3CN, 0-25 o C, 12 h, 80%; e) Et3SiH (6.0 equiv), TFAA (10.0 equiv), TFA (1.0 equiv), CH2Cl2, 0 o C, 3 h, 91% Glycosylation of donor 31 3 with acceptor 3 14,22 to give product 8 cis-4-(1h,1h,2h,2h,3h,3h-perfluoroundecyloxy)-2-butenyl 2,3,4,6-tetra-O-acetyl-β-Dgluccopyranoside (8): Propyl thioglycoside 31 3 (1.48 g, 3.65 mmol) and the alkene-derived fluorous tag 14,22 (1.00 g, 1.82 mmol) were condensed according to the general pentavalent bismuthmediated protocol to give the tag product, 8 (1.39 g, 1.58 mmol). 1 H and 13 C NMR and mass data matched the reported ones above. cis-4-(1h,1h,2h,2h,3h,3h-perfluoroundecyloxy)-2-butenyl 4,6-Obenzylidine-β-D-gluccopyranoside (32): Compound 8 (1.39 g, 1.58 mmol) was dissolved in anhydrous methanol followed by the addition of metallic sodium in bits until the solution was basic to a litmus paper. The reaction was stirred at 0 o C until the reaction 11

12 was completed. The reaction was neutralized with acidic Dowex, filtered and the Dowex rinsed with methanol. The filtrate was concentrated under reduced pressure and co-evaporated with toluene to remove azeotropically remove traces of methanol. The resulting tetra-ol was dried under vacuum for 3 h. The dried material (1.06 g, 1.49 mmol, 1.0 equiv) was dissolved in acetonitrile, after which benzaldehyde dimethyl acetal (454 mg, 2.98 mmol, 2.0 equiv) and PTSA (56.8 mg, 928 µmol, 0.2 equiv) were added. The reaction mixture was stirred at 25 o C until TLC analysis revealed disappearance of all of the starting material. The reaction mixture was neutralized with Et3N, concentrated under reduced pressure and co-evaporated with toluene to azeotropically remove traces of Et3N. The resulting crude material was subjected to MPLC using ISCO (0-50% EtOAc in Hexane) to provide the titled compound 32 (0.920 g, 77%). 1 HNMR (500 MHz, CDCl3) δ (ppm) (m, 2H), 7.36 (dd, J = 5.2, 2.0 Hz, 3H), (m, 2H), 5.53 (s, 1H), 4.43 (d, J = 7.7 Hz, 2H), 4.33 (dd, J = 10.5, 5.0 Hz, 1H), (m, 1H), 4.05 (s, 2H), 3.79 (d, J = 11.6 Hz, 2H), 3.55 (s, 1H), 3.50 (t, J = 6.1 Hz, 3H), 3.44 (td, J = 9.7, 5.0 Hz, 1H), 2.93 (s, 1H), 2.87 (s, 1H), 2.19 (tt, J = 18.6, 8.0 Hz, 2H), (m, 2H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , 80.52, 74.49, 73.20, 68.88, 68.64, 66.47, 65.24, 36.50, 31.47, 28.14, 27.96, 27.79, 20.84, 20.81, HRMS-ESI-TOF (m/z): Calcd. for [C28H27F17O7+Na] , Found cis-4-(1h,1h,2h,2h,3h,3h-perfluoroundecyloxy)-2-butenyl 2,3-di-Obenzyl-4,6-O-benzylidine-β-D-gluccopyranoside (33): The substrate 32 (0.920 g, 1.15 mmol, 1.0 equiv) was dissolved in anhydrous DMF under an argon atmosphere and the resulting solution taken to 0 o C in an ice bath. 60% NaH in an oil suspension (82.9 mg, 3.46 mmol, 3.0 equiv) was added followed by addition of benzyl bromide (591 mg, 3.46 mmol, 3.0equiv). The reaction mixture was warmed up to 25 o C and stirred until the cloudy suspension became clear, indicating completion of the reaction, confirmed by TLC analysis. The reaction was quenched by dropwise addition of MeOH at 0 o C until effervescence ceased. Water was added to the quenched reaction mixture followed by extraction with ethyl acetate (3x). The organic layer was washed with 5% LiCl, dried (Na2SO4) filtered, concentrated under reduced pressure to give the crude material, which was subjected to medium pressure chromatographic purification 12

13 purification via MPLC using ISCO (0-10% EtOAc in Hexane). to afford the product, 33 (0.890 g, 89%). 1 H NMR (500 MHz, CDCl3) δ(ppm) 7.41 (dd, J = 7.6, 2.2 Hz, 2H), (m, 13H), 5.68 (d, J = 5.5 Hz, 2H), 5.49 (s, 1H), 4.82 (t, J = 11.8 Hz, 2H), 4.71 (dd, J = 11.2, 9.4 Hz, 2H), 4.46 (d, J = 7.7 Hz, 1H), (m, 1H), 4.27 (dd, J = 10.5, 5.0 Hz, 1H), (m, 1H), 3.97 (d, J = 5.0 Hz, 2H), (m, 3H), (m, 4H), (m, 2H), (m, 2H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , , , , , 82.05, 81.45, 80.83, 75.33, 75.10, 68.75, 66.49, 66.07, 65.42, 28.19, 27.97, 27.75, HRMS-ESI-TOF (m/z): Calcd. for [C42H39F17O7+Na] , Found cis-4-(1h,1h,2h,2h,3h,3h-perfluoroundecyloxy)-2-butenyl 2,3,6-tri-Obenzyl-β-D-gluccopyranoside (34): To a solution of benzylidinated compound 33 (0.890 g, mmol, 1.0 equiv) in anhydrous dichloromethane at 0 o C under argon was added triethylsilane (0.634 g, 5.46 mmol, 6.0 equiv), trifluoroacetic anhydride (1.04 g, 9.09 mmol, 10 equiv) and trifluoroacetic acid (0.191 g, ml, 1.0 equiv). The reaction mixture was stirred at 0 o C until TLC analysis confirmed a lower running spot (3 h). The reaction mixture was concentrated and co-evaporated with toluene to remove traces of acid. The resulting crude product was purified via FSPE followed by MPLC using ISCO (0-20% EtOAc in Hexane) to furnish the pure compound, 34 (0.891 g, 91%). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 15H), (m, 2H), 4.85 (dd, J = 11.3, 2.2 Hz, 2H), 4.64 (dd, J = 11.3, 4.5 Hz, 2H), (m, 2H), (m, 2H), (m, 1H), (m, 2H), 3.70 (dd, J = 10.4, 3.7 Hz, 1H), 3.62 (dd, J = 10.4, 5.5 Hz, 1H), (m, 1H), (m, 6H), 2.42 (s, 1H), (m, 2H), (m, 2H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , , , , , , , , , , , , , , 84.03, 81.69, 75.30, 74.74, 74.19, 73.67, 71.45, 70.27, 68.73, 66.50, 64.86, 28.19, 28.01, 27.83, 20.86, 20.83, HRMS-ESI-TOF (m/z): Calcd. for [C42H41F17O7+Na] , Found

14 Scheme S3: Synthesis of donors 36 Reagents and Reaction Conditions: BF3.OEt2, PrSH, CH2Cl2, 0-25 o C, 3 h, 79% Propyl 2,3,4,6,2,3,4,6 -hepta-o-acetal-1-thio-β-d-lactopyranoside (36): Compound 35 (2.50 g, 3.68 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane under argon at 0 o C followed by the addition of BF3.OEt2 (1.57 g, 11.1 mmol, 3.0 equiv) dropwise and propanethiol (0.421 g, 5.53 mmol, 1.2). The reaction was taken to room temperature and allowed to stir until TLC analysis unveil complete consumption of the starting material into product (2 h). The reaction mixture was quenched with Et3N, concentrated under reduced pressure to give the crude material, which was purified via medium pressure chromatographic technique via ISCO (using 0-50% EtOAc in Hexane) to afford the pure product, 36 (2.01 g, 79%). 1 H NMR (500 MHz, CDCl3) δ (ppm) 5.33 (dt, J = 3.4, 1.4 Hz, 1H), 5.19 (t, J = 9.2 Hz, 1H), 5.10 (ddd, J = 10.4, 7.9, 6.0 Hz, 1H), (m, 2H), (m, 2H), (m, 2H), (m, 1H), 3.76 (t, J = 9.5 Hz, 1H), 3.60 (ddd, J = 9.9, 5.5, 2.1 Hz, 1H), (m, 2H), 2.14 (d, J = 3.1 Hz, 3H), 2.10 (d, J = 3.9 Hz, 3H), (m, 12H), 1.95 (d, J = 1.7 Hz, 3H), (m, 2H), 0.96 (td, J = 7.3, 1.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , 82.61, 75.77, 75.67, 75.28, 72.79, 69.98, 69.69, 69.39, 68.08, 65.59, 61.29, 61.10, 59.82, 59.80, 31.39, 31.24, 22.12, 21.79, 19.85, 19.82, 19.81, 19.79, 19.77, 19.73, 19.65, 19.64, 19.62, 19.60, 19.50, 19.49, 12.43, HRMS-ESI-TOF (m/z): Calcd. for [C29H42O17S+Na] , Found Glycosylation of donor 41with acceptor 3 to give product 12 Based on the general protocol for pentavalent bismuth-mediated glycosylation of thioglycosides, the donor 36 (127 mg, 182 µmol) and its coupling partner 3 14,22 (100.0 mg, 182 µmol) reacted to 14

15 give the product 12 (164 mg, 77%) after purification via MPLC on ISCO (0-50% EtOAc in Hexane) 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 1H), (m, 1H), 5.32 (dd, J = 3.5, 1.2 Hz, 1H), 5.17 (t, J = 9.3 Hz, 1H), 5.09 (dd, J = 10.4, 7.9 Hz, 1H), 4.94 (dd, J = 10.4, 3.4 Hz, 1H), 4.87 (dd, J = 9.5, 7.9 Hz, 1H), (m, 3H), (m, 1H), (m, 4H), (m, 2H), 3.85 (td, J = 6.8, 6.2, 1.2 Hz, 1H), 3.78 (t, J = 9.4 Hz, 1H), 3.57 (ddd, J = 9.9, 5.1, 2.1 Hz, 1H), 3.46 (t, J = 6.0 Hz, 2H), (m, 2H), 2.13 (s, 3H), 2.10 (s, 3H), (m, 12H), 1.94 (s, 3H), 1.85 (ddd, J = 13.8, 9.9, 6.1 Hz, 2H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , (C-1, JC,H = 165 Hz), 99.36,(C-1, JC,H = 165 Hz) 76.21, 72.85, 72.69, 71.59, 70.96, 70.67, 69.12, 68.74, 66.59, 66.45, 64.75, 61.89, 60.76, 27.95, 20.83, 20.79, 20.76, 20.65, 20.59, 20.57, 20.55, HRMS-ESI-TOF (m/z): Calcd. for [C41H47F17O19+Na] , Found Scheme S4: Synthesis of donor 41, product 13 and acceptor 43 Reagents and conditions: a) (i) HBr, 33 wt % in HOAc (20 equiv), CH2Cl2, 0 25 o C, 4 h, 94%, (ii) CS(NH2)2 (1.5 equiv), CH3CN, Reflux, 10 mins, (iii) Et3N (2.5 equiv), PrBr (1.5 equiv), 25 o C, 2 h, 97% over two steps; b) (i) Na, MeOH, 25 o C, 3 h, 97%, (ii) PhCH(OMe)2 (2.0 equiv), PTSA (0.2 equiv), CH3CN, 25 o C, 4 h, 88%, c) BzCl (1.5 equiv), DMAP (0.1 equiv), Pyridine, 0-25 o C, 2 h, 95%; d) Bu2BOTf (5.0 equiv), BH3.THF (51 equiv), CH2Cl2, 0 o C, 5 h, 86%; e) LevOH (1.0 equiv), DCC (1.0 equiv), DMAP (0.1 equiv), CH2Cl2, 0-25 o C 2 h, 83%. Propyl 3,4,6-tri-O-acetyl-2-deoxy-2-(2,2,2-trichloroacetamido)-β-Dglucopyranoside (38): To a solution of the 1,3,4,6-tetra-O-acetyl-2-deoxy-2-(2,2,2- trichloroacetamido)-β-d-glucopyranose 37 (10.0 g, 20.3 mmol, 1.0 equiv) in anhydrous dichloromethane (50.0 ml) under argon at 0 o C was added HBr solution, 33 wt. % in acetic acid 15

16 (32.9 g, 20.3 mmol, 20.0 equiv). The reaction was taken to and stir at room temperature until TLC analysis (1:2 = EtOAc:Hexane) revealed complete transformation of the starting material into product (4 h), confirmed by a top running spot. The reaction mixture was diluted with dichloromethane, poured into a cold dilute solution of NaHCO3. The two layers were separated and the aqueous layer extracted twice with dichloromethane. The combined organic layers was washed with saturated NaHCO3, brine and water. The organic layer was dried (Na2SO4), filtered, concentrated under reduced pressure and co-evaporated with toluene (3x) to remove traces of HOAc. The crude product was dried under vacuum for 3 h to furnish the intermediate, bromo 3,4,6-tri-O-acetyl-2-deoxy-2-(2,2,2-tichloroacetamido)-α-D-glucopyranoside (9.84 g, 19.1 mmol, 94%), which was then dissolved in anhydrous acetonitrile (50.0 ml) under argon. To this solution was added thiourea (1.75 g, 23.0 mmol, 1.2 equiv) and the resulting reaction mixture reflux for 10 min. The reaction mixture was allowed to cool to room temperature followed by the addition of Et3N (5.82 g, 57.5 mmol, 2.5 equiv) and propyl bromide (2.83 g, 23.0 mmol, 1.2 equiv). The reaction was stirred at 25 o C until TLC analysis confirm an upper running spot. (2 h). The reaction mixture was diluted with dichloromethane and washed with water (2x). The organic layer was dried (Na2SO4), filtered, concentrated under reduced pressure to give the crude material, which was purified via MPLC using ISCO (0-30% EtOAc in Hexane) to afford the desired target, 38 (9.35 g, 97%) 1 H NMR (500 MHz, CDCl3) δ (ppm) 6.98 (d, J = 9.3 Hz, 1H), 5.33 (dd, J = 10.4, 9.4 Hz, 1H), 5.11 (t, J = 9.7 Hz, 1H), 4.66 (d, J = 10.3 Hz, 1H), 4.24 (dd, J = 12.3, 5.2 Hz, 1H), (m, 2H), 3.75 (ddd, J = 10.0, 5.2, 2.4 Hz, 1H), (m, 2H), (m, 9H), 1.62 (qd, J = 7.5, 6.5 Hz, 2H), 0.96 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , 92.20, 84.00, 76.06, 73.12, 68.44, 62.34, 54.78, 32.22, 23.05, 20.73, 20.60, 20.58, HRMS-ESI-TOF (m/z): Calcd. for [C17H24Cl3NO8S+Na] , Found Propyl 4,6-O-benzylidine-2-deoxy-2-(2,2,2-trichloroacetamido)-β-Dglucopyranoside (39): To a solution of the compound 38 in anhydrous methanol was added metallic sodium in bits until the solution was basic to litmus paper. The resulting reaction mixture was stirred at 25 o C until TLC analysis confirmed the transformation of the product. The reaction mixture was quenched by the addition of acidic Dowex. The neutralized 16

17 reaction mixture was filtered through sintered funnel and the Dowex rinse with methanol. This was followed by the concentration of the filtrate under reduced pressure to triol, which was coevaporated with to azeotropically remove methanol. The dried triol (6.85 g, mmol) was dissolved in anhydrous acetonitrile. Benzaldehyde dimethyl acetal (5.45 g, 35.8 mmol, 2.0 equiv), and PTSA (0.681 g, 3.58 mmol) were added and the reaction mixture stirred for 4 h after which TLC (EtOAc:Hexane= 1:1). The reaction mixture was neutralized by the addition of Et3N. The reaction mixture was concentrated under reduced pressure and co-evaporated with toluene to azeotropically remove Et3N. The resulting crude material was subjected to MPLC on ISCO (0-35% EtOAc in dichloromethane) to give the titled product 39 (7.41g, 88%) by using the general procedure for benzylidene formation and MPLC purification on ISCO using 1 HNMR (500 MHz, CDCl3) δ (ppm) 7.41 (dd, J = 6.8, 2.9 Hz, 2H), 7.31 (dd, J = 5.1, 2.0 Hz, 3H), 6.86 (d, J = 7.8 Hz, 1H), 5.48 (s, 1H), 4.85 (d, J = 10.4 Hz, 1H), 4.29 (dd, J = 10.5, 4.5 Hz, 1H), 4.14 (td, J = 9.1, 3.1 Hz, 1H), (m, 2H), (m, 2H), 2.86 (t, J = 1.7 Hz, 2H), (m, 2H), (m, 3H), 0.92 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , 83.63, 81.42, 71.46, 70.42, 68.50, 58.03, 32.56, 23.20, HRMS-ESI-TOF (m/z): Calcd. for [C18H22Cl3NO5S+Na] , Found Propyl 3-O-benzoyl-4,6-O-benzylidine-2-deoxy-2-(2,2,2-trichlorocetamido)- β-d-glucopyranoside (40): Compound 39 (7.41 g, mmol) was dissolved in anhydrous solution of pyridine (60.0 ml) followed by the addition of benzoyl chloride (3.32 g, 23.6 mmol) and DMAP (212 mg, 1.57 mmol) at 0 o C. The reaction mixture was taken to room temperature and stirred until there was complete consumption of the starting material (TLC analysis in 1:2 EtOAc:Hexane). The reaction mixture was poured into a beaker of crushed ice and stirred until all the ice melted. EtOAc (100.0 ml) was added and stirred. The organic layer was separated from the aqueous layer followed by extraction of the aqueous layer with EtOAc (2x100 ml). The combined organic layer was washed with saturated solution of ammonium chloride to remove pyridine. The organic layer was dried (Na2SO4), filtered, concentrated under reduced pressure and co-evaporated with toluene to azeotropically removed traces of pyridine. The resulting crude material was subjected to MPLC purification on ISCO using 0-10% EtOAc in dichloromethane to furnish the titled compound 40 (8.64 g, 95%). 17

18 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 2H), (m, 10H), 5.82 (t, J = 9.9 Hz, 1H), 5.47 (s, 1H), 4.68 (d, J = 10.4 Hz, 1H), (m, 1H), 4.02 (t, J = 4.4 Hz, 1H), 3.84 (t, J = 9.2 Hz, 1H), (m, 2H), (m, 2H), (m, 2H), 0.89 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , 91.17, 83.88, 75.99, 75.73, 72.54, 69.59, 67.31, 53.86, 31.39, 21.90, HRMS-ESI-TOF (m/z): Calcd. for [M+Na] , Found Propyl 3-O-benzoyl-4-O-benzyl-2-deoxy-2-(2,2,2-trichloroacetamido)-β-Dglucopyranoside (41): Dissolution of compound 40 (4.50 g, 7.83 mmol) in BH3- THF complex (38.0 ml, 399 mmol, 51 equiv) at 0 o C was followed by the addition of Bu2BOTf (4.29 g, 15.7 mmol). The reaction mixture was kept and stirred at 0 o C until TLC analysis (1:1 EtOAc:Hexane) confirmed total consumption of the starting material. The reaction mixture was quenched by the addition of Et3N followed by the addition of methanol slowly until effervescence ceased. The reaction mixture was concentrated under reduced pressure and co-evaporated with toluene to azeotropically remove Et3N and methanol and provide the crude material, which was subjected to MPLC purification on ISCO using 0-40% EtOAc in Hexane to yield 4-O-benzylated product 41 (3.87 g, 86%). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 5H), 7.08 (d, J = 9.6 Hz, 1H), 5.33 (dd, J = 10.4, 9.3 Hz, 1H), (m, 2H), 4.52 (d, J = 10.3 Hz, 1H), (m, 1H), (m, 4H), (m, 1H), 3.47 (dtd, J = 9.0, 4.9, 4.4, 2.5 Hz, 1H), (m, 2H), 1.99 (s, 3H), (m, 2H), 1.22 (t, J = 7.0 Hz, 1H), 0.99 (td, J = 7.4, 1.0 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , 84.43, 75.07, 61.53, 54.87, 32.31, 23.01, HRMS-ESI-TOF (m/z): Calcd. for [C25H28Cl3NO6S+Na] , Found Propyl 3-O-benzoyl-4-O-benzyl-6-O-levulinoyl-2-deoxy-2-(2,2,2- trichloroacetamido)-1-thio-β-d-glucopyranoside (42): To a solution of the substrate, 41 (3.87 g, 6.71 mmol, 1.0 equiv) in anhydrous dichloromethane (DCM) at 0 o C were added levulinic acid-levoh (0.887 g, 6.71 mmol, 1.0 equiv), N,N-dicyclohexylcarbodiimide- DCC (1.38 g, 6.71 mmol, 1.0 equiv) and catalytic amount of DMAP The reaction mixture 18

19 was stirred at 25 o C under N2 until TLC revealed complete consumption of the starting material (4 h). To the reaction mixture was added cold saturated solution of NaHCO3 (20.0 ml), followed by extraction with DCM (3x). The combined organic layers was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude material, which was purified via medium pressure liquid chromatography (MPLC) using ISCO (0-35% EtOAC in hexane to give the titled compound 42 (3.87 g, 83%). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 2H), 7.78 (d, J = 9.8 Hz, 1H), 7.60 (d, J = 7.5 Hz, 1H), (m, 2H), 7.19 (dd, J = 5.0, 1.9 Hz, 3H), (m, 2H), 5.86 (dd, J = 10.4, 9.2 Hz, 1H), (m, 3H), 4.35 (q, J = 10.2 Hz, 1H), 4.15 (dd, J = 12.2, 3.5 Hz, 1H), (m, 2H), 3.54 (ddd, J = 9.8, 3.5, 2.1 Hz, 1H), (m, 6H), 2.18 (s, 3H), (m, 2H), 0.96 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , 92.27, 83.88, 75.92, 75.46, 62.45, 54.52, 37.86, 32.01, 29.87, 27.89, 22.98, HRMS-ESI-TOF (m/z): Calcd. for [C30H34Cl3NO8S+Na] , Found Glycosylation of donor 42 with acceptor 21 to give product 13 2-(4-((4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoroundecyl)oxy)phenoxy)ethyl 3-Obenzoyl-4-O-benzyl-6-O-levulinoyl-2-deoxy-2-(2,2,2-trichloroacetamido)-β-Dglucopyranoside (13): By following the pentavalent bismuth-mediated glycosylation protocol, the propyl thioglycoside donor 42 (1.32 g, 1.95 mmol) was couple to the fluorous tag alcohol acceptor 21 (0.600 g, mmol) to provide the tagged product 13 (1.01 g, 85%) after MPLC purification on ISCO using 0-40% EtOAc in Hexane. 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 1H), (m, 10H), 6.73 (d, J = 4.3 Hz, 6H), 5.20 (dd, J = 10.6, 8.7 Hz, 1H), (m, 4H), 4.24 (t, J = 3.2 Hz, 3H), (m, 11H), (m, 3H), (m, 3H), 2.53 (td, J = 6.6, 6.0, 3.6 Hz, 3H), 2.23 (tt, J = 18.4, 7.9 Hz, 3H), 2.12 (s, 4H), 2.00 (dd, J = 10.3, 5.6 Hz, 3H), 1.91 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , , , , , , , (C-1), 19

20 75.99, 75.74, 74.51, 73.92, 72.20, 67.06, 67.04, 65.95, 61.57, 54.57, 36.84, 28.82, 26.85, 19.76, 19.65, HRMS-ESI-TOF (m/z): Calcd. for [C46H41Cl3F17NO11+Na] , Found (4-((4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11- heptadecafluoroundecyl)oxy)phenoxy)ethyl 3-O-benzoyl-4-Obenzyl-2-deoxy-2-(2,2,2-trichloroacetamido)-β-D-glucopyranoside (43): To a solution of the starting material 13 (1.0 equiv) in pyridine/acetic acid (4:1, 15.0 ml) was added hydrazine monohydrate (54.2 mg, 1.08 mmol, 1.3 equiv) After the reaction mixture has been stirred at 25 o C for 30 mins, TLC analysis unveiled complete transformation of the starting material to product. The dilution of the reaction mixture with water, and extraction with EtoAc (3x) gave the combined organic layer, which was washed with saturated solution of ammonium chloride. The organic layer was dried (Na2SO4), concentrated and purified via medium pressure liquid chromatography using 0-40% EtOAc/hexane on ISCO to obtain the product delevulinated product 43 (0.850 g, 92%). 1 H NMR (500 MHz, CDCl3) δ (ppm) 7.92 (dd, J = 8.2, 1.4 Hz, 1H), 7.50 (t, J = 7.5 Hz, 1H), (m, 2H), 7.18 (s, 1H), 7.10 (q, J = 3.4, 3.0 Hz, 2H), 7.03 (dd, J = 6.4, 2.9 Hz, 1H), 6.73 (s, 2H), 5.64 (dd, J = 10.7, 9.1 Hz, 1H), 4.66 (d, J = 8.2 Hz, 1H), (m, 1H), 4.18 (ddd, J = 10.6, 9.5, 8.2 Hz, 1H), (m, 2H), (m, 2H), (m, 2H), 3.51 (td, J = 8.9, 8.3, 4.8 Hz, 1H), 2.23 (tt, J = 18.5, 8.0 Hz, 1H), (m, 2H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , , , , , 92.28, 75.21, 68.49, 67.87, 66.97, 61.41, 55.68, 27.98, HRMS-ESI-TOF (m/z): Calcd. for [C41H35Cl3F17NO9+Na] , Found

21 Glycosylation of donor 36 and 44 with acceptor 34 to give product 14 Donor 36 (142 mg, 204 µmol) and acceptor 34 (100 mg, 102 µmol) were coupled by using the general protocol for pentavalent bismuth-mediated glycosylation of thioglypushcoside to produce the trisaccharide 14 (119 mg, 73%). Likewise, when the donor 44 (149 mg, 204 µmol) was reacted with the acceptor 34 (100 mg, 102 µmol) according to the general glycosylation protocol for thioglycosides, the product 14 (108 mg, 66%) was obtained after purification via MPLC on ISCO (0-50% EtOAc in Hexane) 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 15H), (m, 2H), 5.26 (dd, J = 3.5, 1.1 Hz, 1H), 5.01 (dd, J = 10.4, 7.9 Hz, 1H), (m, 4H), 4.74 (dd, J = 10.0, 8.0 Hz, 2H), 4.67 (dd, J = 11.9, 7.2 Hz, 2H), (m, 2H), (m, 3H), 4.29 (dd, J = 9.7, 7.9 Hz, 2H), (m, 2H), (m, 3H), (m, 2H), (m, 3H), 3.74 (td, J = 6.8, 6.3, 1.3 Hz, 1H), (m, 4H), 3.46 (t, J = 8.9 Hz, 1H), (m, 3H), 3.25 (dt, J = 9.8, 2.8 Hz, 1H), (m, 1H), 2.08 (s, 3H), (m, 2H), 1.97 (s, 3H), 1.95 (d, J = 2.8 Hz, 6H), 1.89 (d, J = 2.5 Hz, 6H), 1.86 (s, 3H), (m, 2H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , , , , , , , , (C-1, JC,H = 162 Hz), (C-1, JC,H = 164 Hz), (C-1, JC,H = 166 Hz), 82.57, 81.60, 75.99, 74.78, 74.65, 74.61, 73.64, 73.19, 72.40, 72.33, 71.00, 70.63, 68.96, 68.70, 67.88, 66.57, 66.53, 64.94, 61.91, 60.75, 29.70, 27.99, 27.81, 20.81, 20.68, 20.62, HRMS-ESI-TOF (m/z): Calcd. for [C68H79NF17O24+NH4] , Found

22 Scheme S5 Synthesis of uronic acid ester donor 49 Reagents and Reaction Conditions: a. (i) Na, MeOH, 4 h, 96%, (ii) Pyridine, TBSCl (1.1 equiv) 3 h, 81%; c. BzCl (4.5 equiv), DMAP (0.01 equiv), Pyridine, 6 h, 86%; d. Et3N.3HF, anhydrous THF, 4 h, 88% e. (i)tempo/baib, CH2Cl2:H2O (2:1) 1 h, (ii) MeI, K2CO3, DMF, 82% Propyl 6-O-tert-butyldimethylsilyl-1-thio-β-D-galactopyranoside (46): The propyl thioglycoside 45 2 (20.0 g, 49.2 mmol) was subjected to the general procedure for deacetylation o give the tetra-ol (11.3 g, 96%). To a portion of the tetra-ol (3.50 g, 14.7 mmol, 1.0 equiv) in anhydrous pyridine (100 ml) was added TBSCl (2.44 g, 16.2 mmol, 1.1 equiv) and the reaction was stirred at 25 o C until all the starting material was consumed to form the product (3 h, TLC analysis). The reaction mixture was poured into a beaker of crushed ice and EtOAc (50 ml) added with stirring. After all the ice has melted, the organic layer was removed and the aqueous layer extracted twice with EtOAc (50 ml). The combined organic layers was washed with a saturated solution of NH4Cl (50 ml) and brine (50 ml), dried (Na2SO4), filtered, and concentrated under reduced pressure to give the crude material which was purified by MPLC, ISCO using 0-50% EtOAc/CH2Cl2 to obtain the product 46 (4.21 g, 81%). 1 H NMR (500 MHz, CDCl3) δ(ppm) 4.28 (d, J = 9.6 Hz, 1H), 4.08 (dd, J = 3.4, 1.1 Hz, 1H), 3.90 (dd, J = 10.5, 5.9 Hz, 1H), 3.85 (dd, J = 10.5, 5.0 Hz, 1H), 3.70 (t, J = 9.3 Hz, 1H), 3.57 (dd, J = 9.0, 3.3 Hz, 1H), 3.50 (d, J = 1.1 Hz, 1H), 2.64 (s, 2H), 2.62 (s, 2H), (m, 2H), 0.99 (t, J = 7.3 Hz, 3H), 0.89 (s, 9H), 0.08 (d, J = 2.1 Hz, 6H). 13 C NMR (126 MHz, CDCl3) δ (ppm) 91.59, 83.71, 82.74, 82.69, 82.49, 82.23, 80.59, 75.99, 74.64, 68.30, 37.46, 31.28, 29.01, 23.70, HRMS-ESI-TOF (m/z): Calcd. For [C15H32O5SSi+Na] , Found Propyl 6-O-tert-butyl-dimethylsilyl-2,3,4-tri-O-benzoyl-1-thio-β-Dgalactopyranoside (47): Compound 46 (4.21 g, 11.9 mmol, 1.0 equiv) dissolved in 22

23 anhydrous pyridine under argon atmosphere. At 0 o C, benzoyl chloride (7.55 g, 53.7 mmol, 4.5 equiv) and DMAP (0.146 g, 1.19 mmol, 0.1 equiv) were added. The reaction mixture was warmed up to room temperature and stirred until TLC analysis indicated completion of the reaction (6 h). The reaction mixture was poured into a beaker of crashed ice followed by the addition of ethyl acetate. After stirring for 15 min, the organic layer was separated and the aqueous layer extracted with ethyl acetate (2x). The combined organic layer was washed with a saturated solution of ammonium chloride, dried (Na2SO4), filtered, concentrated under reduced pressure and the resulting crude material, which furnished the product 47 (6.85 g, 86%) after purification via MPLC on ISCO (0-30% EtOAc in Hexane). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 4H), (m, 2H), (m, 2H), (m, 2H), (m, 3H), 7.50 (td, J = 7.8, 4.1 Hz, 7H), (m, 3H), 6.07 (dd, J = 3.3, 1.1 Hz, 1H), 5.88 (t, J = 9.9 Hz, 1H), 5.72 (dd, J = 9.9, 3.4 Hz, 1H), 4.88 (d, J = 9.9 Hz, 1H), 4.12 (ddd, J = 7.3, 5.9, 1.2 Hz, 1H), 3.93 (dd, J = 10.0, 5.9 Hz, 1H), 3.82 (dd, J = 10.0, 7.6 Hz, 1H), 2.85 (dddd, J = 51.3, 12.4, 8.5, 6.5 Hz, 2H), (m, 2H), 1.04 (t, J = 7.3 Hz, 3H), 0.89 (s, 9H), 0.05 (s, 2H), (s, 2H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , , , 89.78, 83.59, 83.02, 82.96, 82.76, 82.51, 78.83, 74.18, 73.89, 66.69, 37.68, 31.42, 31.39, 28.95, 23.81, 19.21, HRMS-ESI-TOF (m/z): Calcd. for [C36H44O8SSi+Na] , Found Propyl 2,3,4-tri-O-benzoyl-1-thio-β-D-galactopyranoside (41): To solution of compound 47 (2.50 g, 3.76 mmol, 1.0 equiv) in anhydrous THF (30.0 ml) under argon was added Et3N.3HF (1.52 g, 9.40 mmol, 2.5 equiv) at 0 o C. The reaction mixture was stirred at 0 o C until TLC analysis confirmed the formation of the product (4 h). The reaction mixture concentrated under reduced pressure and then co-evaporated with toluene to give the crude mixture, which after purification via MPLC, ISCO, using 0-50% EtOAc/Hexane, gave the targeted product 48 (1.82 g, 88%). 1 HNMR (500 MHz, CDCl3) δ (ppm) (m, 2H), (m, 2H), (m, 2H), (m, 1H), (m, 3H), 7.42 (d, J = 7.8 Hz, 3H), (m, 2H), (m, 2H), 5.65 (dd, J = 10.0, 3.4 Hz, 1H), 4.85 (d, J = 9.9 Hz, 1H), 4.09 (td, J = 6.7, 0.9 Hz, 23

24 1H), 3.87 (dd, J = 12.0, 6.7 Hz, 1H), 3.65 (dd, J = 12.0, 6.8 Hz, 1H), 2.80 (dddd, J = 41.5, 12.5, 8.3, 6.5 Hz, 2H), (m, 2H), 1.00 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ(ppm) , , , , , , , , , , , , , , , 84.32, 77.92, 72.89, 69.24, 68.36, 60.75, 32.18, 23.27, HRMS-ESI-TOF (m/z): Calcd. for [C30H30O8S+Na] , Found Methyl (propyl 2,3,4-tri-O-benzoyl-1-thio-β-D-galactopyranoside) uronate (49): To a vigorously stirred solution of compound 48 (1.82 g 3.31 mmol) in CH2Cl2:H2O (2:1, 30 ml) was added TEMPO (0.103 g, mmol, 0.2 equiv) and BAIB (2.66 g, 8.26 mmol, 2.5 equiv). The stirring was continued until a lower lying spot was observed (TLC, 30 min). The reaction was quenched by the addition of 10% Na2S3O4 solution followed by dilution with water (50 ml) and extraction with ethyl acetate (3x). The combined organic layer was dried (MgSO4), concentrated under reduced pressure and the crude product dried under vacuum. The crude product was dissolved in anhydrous acetonitrile followed by the addition of K2CO3 (1.00 g, 7.27 mmol, 2.2 equiv) and MeI (0.938 g, 6.61 mmol, 2.0 equiv). The reaction mixture was stirred for 12 h, after which TLC analysis (1:2 ethyl acetate:hexane) confirmed the completion of the reaction. The reaction mixture was then diluted with water followed by extraction with ethyl acetate (3x). The combined organic layer was dried (MgSO4), filtered and concentrated under reduced pressure to give the crude product, which was purified via MPLC, ISCO using 0-50% EtOAc/Hexane the product 49 (1.56 g, 82%). 1 H NMR (500 MHz, CDCl3) δ(ppm) (m, 5H), (m, 9H), 6.23 (dd, J = 3.4, 1.3 Hz, 1H), 5.86 (d, J = 9.9 Hz, 1H), 5.67 (dd, J = 10.0, 3.4 Hz, 1H), 4.63 (d, J = 1.3 Hz, 1H), 3.73 (s, 3H), 2.93 (d, J = 32.5 Hz, 2H), (m, 2H), (m, 3H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , 84.04, 76.16, 72.47, 69.39, 67.61, 52.76, 32.06, 23.20, HRMS-ESI-TOF (m/z): Calcd. for [C31H30O9S+Na] , Found

25 Glycosylation of donor 49 with acceptor 30 to give product 15 Donor 49 (117 mg, 203 µmol) was successfully coupled to the acceptor 30 (100 mg, 101 µmol) according to the general glycosylation protocol for thioglycosides to give the product 15 (119 mg, 79%) after purification via MPLC on ISCO (0-25% EtOAC in Hexane). 1 H NMR (500 MHz, CDCl3) δ (ppm) (m, 18H), 7.11 (d, J = 8.0 Hz, 2H), 5.30 (dd, J = 3.5, 1.2 Hz, 1H), 5.20 (dd, J = 10.4, 8.0 Hz, 1H), (m, 4H), 4.77 (d, J = 11.0 Hz, 1H), 4.67 (dd, J = 21.6, 10.9 Hz, 2H), 4.59 (d, J = 12.0 Hz, 1H), (m, 2H), 4.40 (d, J = 7.8 Hz, 1H), (m, 3H), 3.77 (td, J = 6.7, 1.2 Hz, 1H), (m, 2H), (m, 2H), 3.33 (dd, J = 9.9, 8.8 Hz, 1H), (m, 2H), 2.30 (tt, J = 18.1, 8.5 Hz, 2H), 2.08 (s, 3H), 1.94 (s, 3H), 1.91 (d, J = 1.8 Hz, 6H). 13 C NMR (126 MHz, CDCl3) δ (ppm) , , , , , , , , , , , , , , , , , , , , (C-1, JC,H = 165 Hz), (C-1, JC,H = 163 Hz), 84.64, 82.21, 78.00, 75.73, 74.92, 74.89, 74.78, 71.02, 70.68, 70.60, 68.87, 68.43, 67.05, 61.26, 32.93, 29.70, 26.17, 20.83, 20.67, 20.64, HRMS-ESI-TOF (m/z): Calcd. for [C72H65F17O15+NH4] , Found Glycosylation of donor 49 with acceptor 34 to give product (118 mg, 204 µmol) and 34 (100 mg, 102 µmol) were subjected to the general glycosylation protocol for thioglycosides using the pentavalent bismuth to give the product 16 (108 mg, 71%) after purification via MPLC on ISCO using 0-30% EtOAc in Hexane. 1 H NMR (500 MHz, CDCl3) δ (ppm) 7.85 (ddd, J = 15.9, 8.3, 1.4 Hz, 4H), (m, 2H), (m, 4H), (m, 10H), (m, 10H), 5.87 (dd, J = 3.6, 1.4 Hz, 1H), (m, 2H), 5.27 (dd, J = 10.5, 3.5 Hz, 1H), 5.05 (d, J = 11.9 Hz, 1H), (m, 1H), 4.78 (d, J = 11.1 Hz, 1H), 4.60 (d, J = 11.1 Hz, 1H), (m, 2H), 4.10 (t, J =