Synthesis of azido-deoxy and amino-deoxy glycosides and glycosyl fluorides for screening of glycosidase libraries and assembly of substituted glycosides

Article abstract of DOI:10.1016/j.carres.2018.07.007

Azide- and amine-substituted sugars can be useful tools in the probing of biological systems as well as in the assembly of libraries of derivatives using click chemistry or simple amine coupling approaches. A collection of methylumbelliferyl glycosides of

Full text of DOI:10.1016/j.carres.2018.07.007

Accepted Manuscript
Synthesis of azido-deoxy and amino-deoxy glycosides and glycosyl fluorides for
screening of glycosidase libraries and assembly of substituted glycosides
Hong-Ming Chen, Stephen G. Withers
PII:
S0008-6215(18)30359-8
10.1016/j.carres.2018.07.007
CAR 7585
DOI:
Reference:
To appear in: Carbohydrate Research
Received Date: 25 June 2018
Revised Date: 13 July 2018
Accepted Date: 16 July 2018
Please cite this article as: H.-M. Chen, S.G. Withers, Synthesis of azido-deoxy and amino-deoxy
glycosides and glycosyl fluorides for screening of glycosidase libraries and assembly of substituted
glycosides, Carbohydrate Research (2018), doi: 10.1016/j.carres.2018.07.007.
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ACCEPTED MANUSCRIPT

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Synthesis of azido-deoxy and amino-deoxy glycosides and glycosyl fluorides for
screening of glycosidase libraries and assembly of substituted glycosides.
Hong-Ming Chen and Stephen G. Withers
Department of Chemistry,
University of British Columbia
2036 Main Mall
Vancouver, B.C.
Canada V6T 1Z1
Abstract
Azide- and amine- substituted sugars can be useful tools in the probing of biological
systems as well as in the assembly of libraries of derivatives using click chemistry or
simple amine coupling approaches. A collection of methylumbelliferyl glycosides of

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various azido- and amino-deoxy sugar derivatives of glucose, galactose and xylose was
synthesised via azide displacement of the corresponding triflate derivatives and
subsequent modification. These compounds will be used as substrates in a high-
throughput screen to identify glycosidases that can process such modified sugars. The
α-glycosyl fluoride derivatives of each modified sugar were also synthesised to serve as
substrates for glycosynthases derived from the enzymes identified in the screen.
1. Introduction
Azido-sugars have enjoyed substantial usage as reagents for probing cellular
glycosylation events since, in many cases, they can be incorporated onto cell surfaces
using the organism’s own biosynthetic pathways. Once incorporated their locations can
be determined through bio-orthogonal ligation reactions, primarily through click
chemistry with alkyne-containing reagents. Most such studies have been performed with
azide-modified sialic acids or fucose derivatives since these are common terminal
sugars on cell surfaces [1-5].
Applications of azido-sugars are not, however, limited to cell surface labelling. They
have been used in a number of areas, including the assembly of glycopolymers and
dendrimers, in the generation of small molecule libraries and in protein surface
modifications [6]. In a related manner, amino-deoxy sugars are also of interest, both as
naturally occurring entities, such as components of antibiotics or glycosaminoglycans
and themselves as modified, and modifiable sugars (though not bio-orthogonal).
As part of a program to explore the capacity of glycoside hydrolases to accommodate
azido-deoxy and amino-deoxy sugars as substrates we needed access to a range of
fluorogenic glycoside substrates bearing azide substituents at discrete locations around
the ring. These can be used as sensitive potential substrates to screen libraries of
glycosidases for their ability to process such modified sugars [7]. Our hope then is to
generate glycosynthase variants of successful candidate β-glycosidases wherein the
catalytic nucleophile has been replaced by a non-nucleophilic residue. When incubated
with an α-glycosyl fluoride substrate such glycosynthase mutants typically perform
essentially stoichiometric transfer of the glycosyl moiety to a cognate glycosyl acceptor

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[8, 9]. Glycosynthase variants of our target enzymes should then accept their cognate
azido- or amino-deoxy glycosyl fluoride donor sugar and transfer this to suitable
acceptors to assemble azido-sugars containing conjugates under very mild conditions.
In this study, we describe the synthesis of a series of methylumbelliferyl glycosides
bearing azide and amine substituents around the ring. Using common key
intermediates, we further describe the synthesis of a corresponding series of α-glycosyl
fluoride derivatives that will be used as donor sugars for glycosynthases derived from
the candidate glycosidases.
R3
R3
R4
R4
O
O
R2
R1
R2
R1
O
O
O
OH
OH
F
7. R1=OH, R2=N3, R3=H, R4=CH2OH
. R =OH, R =NH , R =H, R =CH OH
4. R1=OH, R2=N3, R3=H, R4=CH2OH
5. R1=OH, R2=NH2, R3=H, R4=CH2OH
12. R1=OH, R2=OH, R3=H, R4=CH2N3
13. R1=OH, R2=OH, R3=H, R4=CH2NH2
21. R1=N3, R2=OH, R3=H, R4=CH2OH
22. R1=NH2, R2=OH, R3=H, R4=CH2OH
29. R1=OH, R2=H, R3=N3, R4=CH2OH
30. R1=OH, R2=H, R3=NH2, R4=CH2OH
38. R1=OH, R2=H, R3=OH, R4=CH2N3
39. R1=OH, R2=H, R3=OH, R4=CH2NH2
8
1
2
2
3
4
2
15. R1=OH, R2=OH, R3=H, R4=CH2N3
16. R1=OH, R2=OH, R3=H, R4=CH2NH2
24. R1=N3, R2=OH, R3=H, R4=CH2OH
25. R1=NH2, R2=OH, R3=H, R4=CH2OH
32. R1=OH, R2=H, R3=N3, R4=CH2OH
33. R1=OH, R2=H, R3=NH2, R4=CH2OH
41. R1=OH, R2=H, R3=OH, R4=CH2N3
42. R1=OH, R2=H, R3=OH, R4=CH2NH2
49. R1=OH, R2=N3, R3=H, R4=H
. R =OH, R =N , R =H, R =H
46
1
2
3
3
4
50. R1=OH, R2=NH2, R3=H, R4=H
47. R1=OH, R2=NH2, R3=H, R4=H
Figure 1. α-D-Glycopyranosyl fluorides and 4-methylumbelliferyl β-D-glycopyranosides
2. Results and Discussion
Introduction of amine and azide substituents into sugar rings typically requires suitably
protection strategy in which the site of substitution is unprotected and of opposite
stereochemistry to the desired product. After activation of the hydroxy to introduce a

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leaving group, substitution by azide is performed followed by introduction of the desired
anomeric substituent. After deprotection, the resulting azido-sugar can be reduced to
the corresponding amine as needed.
After screening a variety of strategies for the synthesis of the azido-deoxy and amino-
deoxy sugar intermediates required we settled on a common approach involving
reaction of the partially protected sugar with triflic anhydride to generate the
corresponding triflate. After a simple work-up, the reactive intermediate was
immediately subjected to azide displacement to generate the azido-deoxy sugar of
inverted configuration at that centre (if stereogenic), typically in net yields of 70-90%.
This protected azide was then converted to its per-O-acetate as necessary to generate
the common intermediate for the two classes of substrate. Subsequent introduction of
the methylumbelliferyl group at the anomeric centre was generally achieved via the
anomeric trichloroacetimidate while the glycosyl fluorides were synthesised from the
per-O-acetate using HF/pyridine. Reduction of the deprotected azido-glycosyl fluorides
was achieved via catalytic hydrogenation, while reduction of the deprotected
methylumbelliferyl azido-sugars required use of triphenylphosphine to avoid partial
reduction of the methyumbelliferyl group.
4-Azido-4-deoxy and 4-amino-4-deoxy glucose derivatives were synthesised from the
readily available 1,2,3,6-tetra-O-benzoyl-D-galactose as shown in Scheme 1.
Conversion to the 4-azido-glucose derivative proceeded well via the triflate, with silica
gel clean-up prior to Zemplen deprotection and per-O-acetylation in a total yield (over 4
steps) of 52%.
6-Azido-6-deoxy and 6-amino-deoxy glucose derivatives were generated by selective
triflation of the 6-hydroxyl of 1,2,3-tri-O-acetyl-D-glucopyranose at low temperature,
followed by azide displacement and then acetylation of the 4-hydroxyl using acetic
anhydride in pyridine (Scheme 2). This yielded the per-O-acetate directly in a combined
yield of 57%.
3-Azido-3-deoxy and 3-amino-deoxy glucose derivatives were generated from 1,2:5,6-
di-O-cyclohexylidene-D-allofuranose [10, 11] by sequential triflation and azide

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displacement (Scheme 3). Deprotection using aqueous trifluoroacetic acid (TFA)
followed by acetylation using acetic anhydride and pyridine yielded the per-O-acetate of
the 3-azido-sugar in a combined yield of 75%.
4-Azido-4-deoxy and 4-amino-deoxy galactose derivatives were synthesised from 1,2,3-
tri-O-acetyl-D-glucopyranose by selective acetylation of the 6-hydroxyl using 1-
(acetoxy)-benzotriazole followed by triflation of the free 4-hydroxyl and displacement
with azide (Scheme 4). This yielded the per-O-acetate of 4-azido-4-deoxy-D-galactose
directly in a combined yield of 72%.
6-Azido-6-deoxy and 6-amino-deoxy galactose derivatives were generated from 1,2:3,4-
di-O-isopropylidene-α- -galactopyranose by triflation of the 6 hydroxyl at 0 °C followed
D
by azide displacement in dry DMF (Scheme 5). Deprotection with aqueous TFA
followed by acetylation with acetic anhydride and pyridine yielded the per-O-acetate of
6-azido-6-deoxy-D-galactose in a total yield of 69%.
4-Azido-4-deoxy and 4-amino-deoxy xylose derivatives were synthesised from
L
-
arabinose by an initial selective benzoylation of the 1,2 and 3-hydroxyls using benzoyl
chloride in pyridine at 0 °C, followed by triflation of the axial 4 hydroxyl and sodium
azide displacement (Scheme 6). Deprotection under Zemplen conditions was followed
by acetylation with acetic anhydride and pyridine to yield the per-O-acetate of 4-azido-4-
deoxy-D-xylose in a total yield of 33% from L-arabinose.
3. Experimental
General Information
All chemicals were of analytical grade purchased from the Sigma-Aldrich company,
unless otherwise stated. All solvents were BOC standard grade and distilled before use.
Dichloromethane and pyridine were distilled from calcium hydride. Methanol was
distilled from magnesium. THF was distilled from sodium. DMF was dried and stored
over 4 Å molecular sieves. Analytical thin-layer chromatography (TLC) was performed
on aluminium-backed sheets of Silica Gel 60F254 (E. Merck) of thickness 0.2 mm. The

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plates were visualised using UV light (254 nm) and/or by exposure to 10% ammonium
molybdate (2 M in H2SO4) followed by charring. Flash column chromatography was
carried out using Silicycle silica gel (230-400 mesh). Proton and carbon NMR spectra
were recorded on Bruker Advance 400inv, 400dir and 300 Fourier Transform
spectrometer fitted with 5 mm BBI-Z probe. All spectra were recorded using an internal
deuterium lock and are referenced internally using the residual solvent peak. Carbon
and proton chemical shifts are quoted in parts per million (ppm) downfield of
tetramethylsilane, fluorine chemical shifts are quoted downfield of trifluoroacetic acid
(TFA). Coupling constants (J) are given in Hertz (Hz). Carbon NMR spectra were
performed with broadband proton decoupling and were recorded with DEPT. Mass
spectra were recorded on a Waters/Micromass LCT using electrospray ionisation (ESI)
and recorded using Time-Of-Flight (TOF) method using methanol as solvent.
General Procedures
A. Fluorination [12]
A solution of acetylated azido-sugar (5 mmol) in dry CH2Cl2 (10 mL) was transferred to
a 100 mL plastic bottle, cooled to 0 °C, then HF/pyridine (15 mL, 70% HF) was added.
The plastic bottle was capped and stirred overnight at 0 °C. The reaction mixture was
slowly poured into a plastic beaker with ice, saturated NaHCO3 and solid NaHCO3
cooled with an ice bath with stirring. The crude product was extracted with EtOAc (2 x
100 mL), washed with saturated NaHCO3 (2 x 100 mL) and brine (100 mL), dried over
MgSO4, filtered and concentrated. The resulting residue was purified by flash column
chromatography (4:1, petroleum ether-EtOAc).
B. Deacetylation with NH3 [12]
To a solution of acetylated azido-glycosyl fluoride (2 mmol) in dry methanol (50 mL) was
slowly bubbled ammonia for 5 minutes at 0 °C, and the reaction mixture was stirred at
0 °C until completion. The solvent was evaporated, and the resulting residue was
purified by flash column chromatography (9:1, CH2Cl2-MeOH).

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C. Reduction of azide with Pd/C [12]
A mixture of azido-glycosyl fluoride (3 mmol), Pd/C (10% on activated charcoal, 200
mg) and methanol (50 mL) was evacuated and refilled with H2 three times, and stirred
overnight at room temperature under an atmosphere of hydrogen. The reaction mixture
was filtered through a short pad of Celite and washed with methanol, and then the
solvent was evaporated. The resulting residue was purified by flash column
chromatography (10:2:1, EtOAc-MeOH-1 M NH4OH or 5:1, CH2Cl2-MeOH with 1%
Et3N).
D. Anomeric deacetylation [13, 14]
A solution of azido-D-glucose peracetate (3.0 mmol) and hydrazine acetate (360 mg, 3.9
mmol, 1.3 equiv) in DMF (11 mL) was stirred for 3 h at room temperature, diluted with
EtOAc (100 mL), washed with H2O (2 x 50 mL), saturated NaHCO3 (50 mL) and brine
(50 mL), dried over MgSO4, filtered and concentrated. The product was purified by flash
column chromatography.
E. Glycosylation [15, 16]
To a solution of acetylated azido-sugar hemiacetal (2 mmol) and trichloroacetonitrile
(TCA, 2 mL) in dry CH2Cl2 (20 mL) was added 1,8-diazabicyclo[5.4.0]-undec-7-ene
(DBU, 20 µL) at -20 °C with stirring under N2, and the mixture was gradually warmed to
0 °C. When TLC showed the reaction was complete, the solvent was evaporated under
reduced pressure. The resulting residue was purified by passage through a silica gel
plug (3:1, petroleum ether-EtOAc) to afford a colorless syrup. To the syrup were added
dry CH2Cl2 (25 mL), 4-methyl-7-hydroxylcoumarin (Mu, 2 equiv) and 4 Å molecular
sieves, and the mixture was stirred for 10 minutes at room temperature under N2. The
mixture was then cooled to -40 °C, BF3 Et2O (20 µL) was added dropwise and the
reaction mixture was stirred for 5 h under the same conditions until completion. After
quenching by addition of Et3N, the mixture was filtered through a short pad of Celite and
washed with CH2Cl2. The filtrate was washed with 1 M NaOH (3 x 50 mL), saturated
NaHCO3 (50 mL) and brine (50 mL), dried over MgSO4, filtered and concentrated. The

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product was purified by flash column chromatography (2:1, petroleum ether-EtOAc) to
afford a white solid.
F. Deacetylation with sodium [17, 18]
A solution of acetylated 4-methylumbelliferyl azido-glycoside (1 mmol) in dry methanol
(60 mL) was treated with a catalytic amount of sodium under N2 at room temperature.
The white precipitate was filtered and washed with methanol. The mother liquor was
neutralized with Amberlite 120 H, filtered and concentrated. The product was
precipitated out with methanol and diethyl ether. The combined white solid was dried
under vacuum.
G. Reduction of azide with triphenylphosphine [19, 20]
A mixture of acetylated 4-methylumbelliferyl azido-glycoside (0.3 mmol), Ph3P (110 mg,
0.4 mmol, 1.4 equiv), silica gel (230~400 mesh, 300 mg) and THF/H2O (4/1, 14 mL) was
stirred overnight at 50 °C. After completion, the solvents were removed under reduced
pressure. The product was purified by flash column chromatography (17:2:1 and 10:2:1,
EtOAc/MeOH/1 M NH4OH).

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Scheme 1. Reagents and conditions: a) CH2Cl2, pyridine, Tf2O, 0 °C; DMF, NaN3, rt;
MeOH, NaOMe, rt; pyridine, Ac2O, rt; 52% (four steps). b) CH2Cl2, HF/pyridine, 0 °C,
73%. c) MeOH, NH3 (g), 0 °C, 95%. d) MeOH, Pd/C, H2, rt, 98%. e) DMF, hydrazine
acetate, rt; CH2Cl2, TCA, DBU, -20~0 °C; CH2Cl2, 4 Å molecular sieves, Mu, BF3 Et2O, -
40~-30 °C; 52%. f) MeOH, Na, rt, 91%. g) THF/H2O (4/1), Ph3P, silica gel, 50 °C, 98%.
4-Azido-4-deoxy-1,2,3,6-tetra-O-acetyl-D-glucopyranose (2): To a solution of 1 [21,
22] (16 g, 26.9 mmol) in dry CH2Cl2 (160 mL) and dry pyridine (20 mL), stirred at 0 °C
under N2, was added dropwise triflic anhydride (Tf2O, 9.6 mL, 57.1 mmol, 2.1 equiv)
[23]. After addition, the reaction mixture was stirred for 1.5 h under the same conditions,
diluted with CH2Cl2 (150 mL), washed with cold 1 M HCl (3 x 100 mL), cold saturated
NaHCO3 (2 x 100 mL) and brine (100 mL), dried over MgSO4, filtered and concentrated
to afford a yellow foam. To the yellow foam were added dry DMF (100 mL) and NaN3
(15 g, 230.8 mmol, 8.6 equiv) at room temperature under N2, and the suspension was
stirred vigorously for 4 h at the same condition. The reaction mixture was diluted with
EtOAc (600 mL), washed with water (2 x 300 mL) and brine (300 mL), dried over

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MgSO4, filtered and concentrated. The resulting residue was purified over a silica gel
plug (4:1 petroleum ether-EtOAc) to afford 4-azido-4-deoxy-1,2,3,6-tetra-O-benzoyl-D-
glucose [24] as a white foam. To a solution of the white foam in dry methanol (360 mL)
was added sodium methylate solution in MeOH (3 mL, 5.4 M), and the reaction mixture
was stirred overnight at room temperature, neutralized with Amberlite 120 H, filtered,
washed with methanol and concentrated to give crude 4-azido-4-deoxy-D-glucose.
This material was acetylated with pyridine (75 mL) and acetic anhydride (50 mL) at
room temperature overnight. After evaporation under reduced pressure, water (50 mL)
was added to the residue, and the mixture was stirred for 0.5 h at room temperature,
extracted with EtOAc (2 x 200 mL). The organic phase was washed with 1 M HCl (2 x
200 mL), saturated NaHCO3 (3 x 200 mL) and brine (200 mL), dried over MgSO4,
filtered and concentrated. The product was purified by flash column chromatography
(3:1 petroleum ether-EtOAc) to afford 2 as a colorless syrup (5.21 g, 52% overall yield).
1H NMR showed the ratio of α/β is about 1/1. ESI-MS: Calcd for [C14H19N3O9 + Na]+:
396.1; Found m/z: 396.3.
4-Azido-4-deoxy-2,3,6-tri-O-acetyl-α-
D
-glucopyranosyl fluoride (3) [12]: See general
1
procedure A, 3 was obtained as a colorless syrup in 79% yield. H NMR (CDCl3, 400
MHz):
δ 5.75 (dd, 1 H, J1,2 2.4 Hz, JH,F 52.4 Hz, H-1), 5.54 (t, 1 H, H-3), 4.96 (dddd, 1 H,
J2,3 10.0 Hz, JH,F 23.7 Hz, H-2), 4.47 (dd, 1 H, J5,6a 2.4 Hz, J6a,6b 12.4 Hz, H-6a), 4.34
(dd, 1 H, J5,6b 4.0 Hz, H-6b), 4.00 (m, 1 H, H-5), 3.74 (t, 1 H, J3,4 = J4,5 10.4 Hz, H-4),
2.18 (s, 3 H, CH3CO), 2.17 (s, 3 H, CH3CO), 2.15 (s, 3 H, CH3CO). 13C NMR (CDCl3, 75
MHz):
δ 170.5, 170.3, 169.7, 104.1 (d, JC,F 228.4 Hz), 70.33 (d, JC,F 24.3 Hz), 70.31 (d,
JC,F 3.8 Hz), 70.1, 62.2, 59.5, 20.9, 20.8, 20.7. 19F-NMR (CDCl3, 282 MHz):
δ -149.4.
ESI-HRMS: Calcd for [C12H16N3FO7 + Na]+: 356.0870; Found m/z: 356.0861.
4-Azido-4-deoxy-α-D-glucopyranosyl fluoride (4): See general procedure B, the
mixture was stirred for 2 days at 0 °C. The resulting residue was purified by flash
column chromatography to afford 4 as a colorless syrup in 94% yield. 1H NMR (CD3OD,
400 MHz):
6), 3.50 (dddd, 1 H, J2,3 10.0 Hz, JH,F 28.8 Hz, H-2). 13C NMR (CD3OD, 100 MHz):
107.7 (d, JC,F 223.0 Hz), 73.1 (d, JC,F 3.0 Hz), 72.3, 71.8 (d, JC,F 24.0 Hz), 61.3, 60.6.
δ 5.58 (dd, 1 H, J1,2 2.8 Hz, JH,F 53.6 Hz, H-1), 3.80~3.49 (m, 5 H, H-3, 4, 5 &
δ

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19F-NMR (CD3OD, 282 MHz):
206.0577; Found m/z: 206.0580.
δ
-152.1. ESI-HRMS: Calcd for [C6H10N3FO4 - H]-:
4-Amino-4-deoxy-α-
obtained as a white foam in 98% yield. 1H NMR (CD3OD, 400 MHz):
D
-glucopyranosyl fluoride (5): See general procedure C, 5 was
5.61 (dd, 1 H, J1,2
δ
2.4 Hz, JH,F 53.2 Hz, H-1), 3.52 (m, 1 H, H-5), 3.38 (m, 3 H, H-3 & H-6), 3.11 (dddd, 1 H,
13
J2,3 9.2 Hz, JH,F 26.0 Hz, H-2), 2.71 (t, J3,4 = J4,5 10.0 Hz, H-4). C NMR (CD3OD, 100
MHz):
δ 109.8 (d, JC,F 224.0 Hz), 74.0 (d, JC,F 24.0 Hz), 73.8 (d, JC,F 4.0 Hz), 71.8, 63.0,
54.8. 19F-NMR (CD3OD, 282 MHz):
δ
-151.9. ESI-HRMS: Calcd for [C6H12NFO4 + H]+:
182.0829; Found m/z: 182.0826.
4-Methylumbelliferyl 4-azido-4-deoxy-2,3,6-tri-O-acetyl-β-
See general procedure D, eluent (3:1 & 2:1 petroleum ether-ethyl acetate), 4-azido-4-
deoxy-2,3,6-tri-O-acetyl- -glucopyranose was obtained as a colorless syrup in 74%
D-glucopyranoside (6):
D
yield. See general procedure E, eluent (2:1 petroleum ether-EtOAc), 6 was obtained as
1
a white solid in 70% yield (overall yield: 52%). H NMR (d6-acetone, 400 MHz):
δ 7.71
(d, 1 H, J 8.4 Hz, Ar-H), 7.01 (m, 2 H, Ar-H), 6.19 (d, 1 H, J 1.2 Hz, Ar-H), 5.59 (d, 1 H,
J1,2 7.6 Hz, H-1), 5.42 (t, 1 H, J2,3 = J3,4 9.6 Hz, H-3), 5.19 (dd, 1 H, H-2), 4.48 (dd, 1 H,
J5,6a 2.4 Hz, J6a,6b 12.0 Hz, H-6a), 4.36 (dd, 1 H, J5,6b 5.6 Hz, H-6b), 4.14 (dddd, 1 H, H-
5), 3.99 (t, 1 H, J4,5 10.0 Hz, H-4), 2.44 (d, J 1.2 Hz, 3 H, Ar-CH3), 2.10 (s, 3 H, CH3CO),
2.09 (s, 3 H, CH3CO), 2.03 (s, 3 H, CH3CO). 13C NMR (d6-acetone, 100 MHz):
169.5, 169.2, 159.8, 159.6, 155.1, 152.7, 126.5, 115.4, 113.5, 112.7, 103.7, 97.8, 73.3,
δ 170.0,
72.4, 71.2, 62.6, 60.0, 19.9, 19.88, 19.8, 17.8. ESI-HRMS: Calcd for [C22H23N3O10
Na]+: 512.1281; Found m/z: 512.1277.
+
4-Methylumbelliferyl 4-azido-4-deoxy-β-
procedure F, 7 was obtained as a white solid in 91% yield. H NMR (d6-DMSO + D2O,
400 MHz): 7.72 (d, 1 H, J 9.6 Hz, Ar-H), 7.04 (m, 2 H, Ar-H), 6.26 (d, 1 H, J 1.2 Hz, Ar-
D-glucopyranoside (7): See general
1
δ
H), 5.13 (d, 1 H, J1,2 8.0 Hz, H-1), 3.64~3.42 (m, 5 H, H-2, 3, 5 & 6), 3.35 (t, 1 H, J 3,4
=
13
J4,5 8.4 Hz, H-4), 2.41 (s, 3 H, Ar-CH3). C NMR (d6-DMSO + D2O, 100 MHz):
δ 160.7,
160.5, 155.0, 153.9, 127.1, 114.8, 113.9, 112.4, 103.7, 100.0, 75.8, 75.0, 73.8, 62.1,
61.0, 18.7. ESI-HRMS: Calcd for [C16H17N3O7 + Na]+: 386.0964; Found m/z: 386.0956.

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4-Methylumbelliferyl 4-amino-4-deoxy-β-
procedure G, 8 was obtained as a white solid in 98% yield. H NMR (d6-DMSO + D2O,
400 MHz): 7.68 (d, 1 H, J 8.4 Hz, Ar-H), 7.03 (m, 2 H, Ar-H), 6.21 (s, 1 H, Ar-H), 5.00
D-glucopyranoside (8) [25]: See general
1
δ
(d, 1 H, J1,2 7.6 Hz, H-1), 3.67 (dd, 1 H, J5,6a 2.4 Hz, J6a,6b 12.0 Hz, H-6a), 3.50 (dd, 1 H,
J5,6b 5.2 Hz, H-6b), 3.40 (m, 1 H, H-5), 3.29 (t, 1 H, H-2), 3.21 (t, 1 H, J2,3 = J3,4 9.2 Hz,
13
H-3), 2.54 (t, 1 H, J4,5 9.6 Hz, H-4), 2.38 (s, 3 H, Ar-CH3). C NMR (d6-DMSO + D2O,
100 MHz):
δ 160.8, 160.7, 154.9, 154.1, 127.0, 114.7, 114.0, 112.2, 103.7, 100.7, 77.8,
76.6, 73.7, 61.6, 53.8, 18.7. ESI-HRMS: Calcd for [C16H19NO7 + H]+: 338.1240; Found
m/z: 338.1234.
Scheme 2. Reagents and conditions: a) CH2Cl2, pyridine, Tf2O, -78~-30 °C; DMF,
NaN3, rt; pyridine, Ac2O, rt;.57%. b) CH2Cl2, HF/pyridine, 0 °C, 75%. c) MeOH, NH3 (g),
0 °C, 95%. d) MeOH, Pd/C, H2, rt, 98%. e) DMF, hydrazine acetate, rt; CH2Cl2, TCA,
DBU, -20~0 °C; DCM, 4 Å molecular sieves, Mu, BF3 Et2O, -40~-30 °C; 51%. f) MeOH,
Na, rt, 96%. g) THF/H2O (4/1), Ph3P, silica gel, 50 °C, 93%.

ACCEPTED MANUSCRIPT
13
6-Azido-6-deoxy-1,2,3,4-tetra-O-acetyl-
D
-glucopyranose (10): To a solution of 1,2,3-
tri-O-acetyl-
D
-glucopyranose [26, 27] (9, 0.58 g, 1.9 mmol) in dry CH2Cl2 (36 mL) and
dry pyridine (2 mL), stirred at -78 °C under N2, was added triflic anhydride (0.5 mL, 3.0
mmol, 1.6 equiv) dropwise. After addition, the dry-ice cooling bath was gradually
warmed up to -30 °C, and TLC showed completion of reaction. The reaction mixture
was diluted with CH2Cl2 (30 mL), washed with cold 1 M HCl (30 mL), cold saturated
NaHCO3 (30 mL) and cold brine (30 mL), dried over MgSO4, filtered and evaporated,
dried under vacuum to afford a brownish foam. To the brownish foam were added dry
DMF (10 mL) and NaN3 (1.24 g, 19.1 mmol, 10 equiv), and the mixture was stirred
vigorously overnight at room temperature under N2. The reaction mixture was diluted
with EtOAc (100 mL), washed with water (50 mL) and brine (50 mL), dried over MgSO4,
filtered and evaporated to afford crude 6-Azido-6-deoxy-1,2,3-tri-O-acetyl-D-
glucopyranose. To the residue were added pyridine (5 mL) and acetic anhydride (2.5
mL), and the mixture was stirred overnight at room temperature. After evaporation
under reduced pressure, the residue was dissolved in EtOAc (60 mL), washed with 1 M
HCl (30 mL), saturated NaHCO3 (2 x 30 mL) and brine (30 mL), dried over MgSO4,
filtered and concentrated. The residue was purified by flash column chromatography
(2:1 petroleum ether-EtOAc) to afford 10 [28] as a colorless syrup (57%; α/β = 1/1.4). 1H
NMR (CDCl3, 400 MHz, α-anomer): δ 6.38 (d, 1 H, J1,2 3.6 Hz, H-1), 5.49 (t, 1 H, H-3),
5.13 (t, 1 H, J3,4 = J4,5 9.8 Hz, H-4), 5.12 (dd, 1 H, J2,3 10.4 Hz, H-2), 4.11 (dddd, 1 H, H-
5), 3.43 (dd, 1 H, J5,6a 2.8 Hz, J6a,6b 13.6 Hz, H-6a), 3.33 (dd, 1 H, J5,6b 5.2 Hz, H-6b),
2.22 (s, 3 H, CH3CO), 2.09 (s, 3 H, CH3CO), 2.06 (s, 3 H, CH3CO), 2.05 (s, 3 H,
CH3CO). ESI-MS: Calcd for [C14H19N3O9 + Na]+: 396.1; Found m/z: 396.5.
6-Azido-6-deoxy-2,3,4-tri-O-acetyl-α-
D
-glucopyranosyl fluoride (11): See general
1
procedure A, 11 was obtained as a white solid in 75%. H NMR (CDCl3, 400 MHz):
δ
5.78 (dd, 1 H, J1,2 2.8 Hz, JH,F 52.8 Hz, H-1), 5.49 (t, 1 H, H-3), 5.14 (t, 1 H, J3,4 = J4,5 9.8
Hz, H-4), 4.95 (dddd, 1 H, J2,3 10.4 Hz, JH,F 24.0 Hz, H-2), 4.15 (dddd, 1 H, H-5), 3.46
(dd, 1 H, J5,6a 2.4 Hz, J6a,6b 13.6 Hz, H-6a), 3.33 (dd, 1 H, J5,6b 5.2 Hz, H-6b), 2.11 (s, 3
H, CH3CO), 2.05 (s, 3 H, CH3CO), 2.03 (s, 3 H, CH3CO). 13C NMR (CDCl3, 100 MHz):
170.1, 170.06, 170.5, 103.7 (d, JC,F 228.0 Hz), 70.9 (d, JC,F 4.0 Hz), 70.3 (d, JC,F 24.0
δ

ACCEPTED MANUSCRIPT
14
19
Hz), 69.3, 68.5, 50.7, 20.73, 20.69, 20.65. F-NMR (CDCl3, 282 MHz):
δ -149.5. ESI-
HRMS: Calcd for [C12H16FN3O7 + Na]+: 356.0870; Found m/z: 356.0865.
6-Azido-6-deoxy-α-
obtained as a white solid in 95% yield. 1H NMR (D2O, 400 MHz):
Hz, JH,F 53.2 Hz, H-1), 3.86 (dddd, 1 H, J4,5 9.6 Hz, J5,6a 2.4 Hz, J5,6b 5.2 Hz, H-5), 3.70 ~
3.42 (m, 5 H, H-2, 3, 5 & 6). 13C NMR (D2O, 100 MHz):
107.3 (d, JC,F 222.0 Hz), 73.0
D
-glucopyranosyl fluoride (12): See general procedure B, 12 was
δ
5.60 (dd, 1 H, J1,2 2.8
δ
19
(d, JC,F 3.0 Hz), 72.3, 71.0 (d, JC,F 25.0 Hz), 69.4, 50.7. F-NMR (D2O, 282 MHz):
δ -
150.4. ESI-HRMS: Calcd for [C6H10N3FO4 - H]-: 206.0577; Found m/z: 206.0585.
6-Amino-6-deoxy-α-D-glucopyranosyl fluoride (13): See general procedure C, 13
1
was obtained as a white solid in 98% yield. H NMR (D2O, 400 MHz):
δ
5.59 (dd, 1 H,
J1,2 2.8 Hz, JH,F 53.2 Hz, H-1), 3.72 (m, 1 H, H-5), 3.62 (t, 1 H, H-3), 3.52 (dddd, 1 H, J2,3
9.6 Hz, JH,F 26.0 Hz, H-2), 3.32 (t, 1 H, J3,4 = J4,5 9.6 Hz, H-4), 3.00 (dd, 1 H, J5,6a 2.6 Hz,
H-6a), 2.78 (dd, 1 H, J5,6b 7.2 Hz, J6a,6b 13.6 Hz, H-6b). 13C NMR (D2O, 100 MHz):
δ
107.3 (d, JC,F 221.0 Hz), 73.6 (d, JC,F 3.0 Hz), 72.4, 71.2 (d, JC,F 25.0 Hz), 70.2, 41.0.
19F-NMR (D2O, 282 MHz):
Found m/z: 182.0824.
δ
-150.0. ESI-HRMS: Calcd for [C6H12NFO4 + H]+: 182.0829;
4-Methylumbelliferyl 6-azido-6-deoxy-2,3,4-tri-O-acetyl-β-
See general procedure D, eluent (3:1 & 2:1 petroleum ether-EtOAc), 6-azido-6-deoxy-
2,3,4-tri-O-acetyl- -glucopyranose was obtained as a colorless syrup in 85% yield.
D-glucopyranoside (14):
D
See general procedure E, eluent (2:1 & 3:2 petroleum ether-EtOAc), 14 was obtained
1
as a white solid in 60% yield (overall yield: 51%). H NMR (d6-acetone, 400 MHz):
δ
7.73 (d, 1 H, J 9.2 Hz, Ar-H), 7.05 (m, 2 H, Ar-H), 6.20 (d, 1 H, J 1.2 Hz, Ar-H), 5.67 (d,
1 H, J1,2 8.0 Hz, H-1), 5.42 (t, 1 H, J3,4 9.6 Hz, H-3), 5.26 (dd, 1 H, J2,3 9.6 Hz, H-2), 5.13
(t, 1 H, J4,5 10.0 Hz, H-4), 4.29 (dddd, 1 H, H-5), 3.62 (dd, 1 H, J5,6a 2.8 Hz, J6a,6b 13.6
Hz, H-6a), 3.53 (dd, 1 H, J5,6b 6.8 Hz, H-6b), 2.45 (d, J 1.2 Hz, 3 H, Ar-CH3), 2.04 (s, 3
H, CH3CO), 2.03 (s, 3 H, CH3CO), 1.99 (s, 3 H, CH3CO). 13C NMR (d6-acetone, 100
MHz):
δ 169.6, 169.3, 169.0, 159.8, 159.6, 155.1, 152.7, 126.6, 115.5, 113.5, 112.8,
103.7, 97.9, 73.3, 72.4, 70.9, 69.2, 51.0, 19.9, 19.8 (2 C), 17.8. ESI-HRMS: Calcd for
[C22H23N3O10 + Na]+: 512.1281; Found m/z: 512.1288.

ACCEPTED MANUSCRIPT
15
4-Methylumbelliferyl 6-azido-6-deoxy-β-
procedure F, 15 was obtained as a white solid in 96% yield. H NMR (d6-DMSO + D2O,
400 MHz): 7.70 (d, 1 H, J 8.8 Hz, Ar-H), 7.05 (m, 2 H, Ar-H), 6.24 (d, 1 H, J 0.8 Hz, Ar-
D-glucopyranoside (15): See general
1
δ
H), 5.16 (d, 1 H, J1,2 7.2 Hz, H-1), 3.72 (m, 1 H, H-5), 3.50~3.40 (m, 2 H, H-6),
3.34 3.27 (m, 2 H, H-2 & H-3), 3.14 (t, 1 H, J3,4 = J4,5 9.0 Hz, H-4), 2.40 (s, 3 H, Ar-CH3).
13C NMR (d6-DMSO + D2O, 100 MHz):
δ 160.8, 160.4, 155.0, 154.0, 127.0, 114.8,
114.0, 112.3, 103.7, 100.1, 76.3, 75.8, 73.5, 71.0, 51.9, 18.7. ESI-HRMS: Calcd for
[C16H17N3O7 + Na]+: 386.0964; Found m/z: 386.0953.
4-Methylumbelliferyl 6-amino-6-deoxy-β-
D
-glucopyranoside (16): See general
1
procedure G, 16 was obtained as a white solid in 93% yield. H NMR (d6-DMSO + D2O,
400 MHz):
δ 7.69 (d, 1 H, J 8.8 Hz, Ar-H), 7.03 (m, 2 H, Ar-H), 6.23(d, 1 H, J 0.8 Hz, Ar-
H), 5.04 (d, 1 H, J1,2 7.2 Hz, H-1), 3.42 (m, 1 H, H-5), 3.29 (m, 2 H, H-2 & H-3), 3.12 (t, 1
H, J3,4 = J4,5 9.2Hz, H-4), 2.91 (dd, 1 H, J5,6a 2.4 Hz, J6a,6b 13.2 Hz, H-6a), 2.64 (dd, 1 H,
J5,6b 7.2 Hz, H-6b), 2.39 (d, 3 H, J 1.4 Hz, Ar-CH3). 13C NMR (d6-DMSO + D2O, 100
MHz):
δ 160.7, 160.6, 155.0, 153.9, 127.1, 114.7, 113.8, 112.3, 103.7, 100.5, 77.2,
76.9, 73.8, 71.8, 43.2, 18.7. ESI-HRMS: Calcd for [C16H19NO7 + Na]+: 360.1059; Found
m/z: 360.1050.

ACCEPTED MANUSCRIPT
16
Scheme 3. Reagents and conditions: a) CH2Cl2, DMP, 0 °C~rt; EtOH, H2O, NaBH4, 0
°C~rt; CH2Cl2, pyridine, Tf2O, 0 °C; DMF, NaN3, rt, 96%. b) TFA, H2O, rt; pyridine, Ac2O,
rt, 78%. c) CH2Cl2, Ac2O, HF/pyridine, 0 °C, 76%. d) MeOH, NH3, 0 °C, 97%. e) MeOH,
Pd/C, H2, rt, 96%. f) DMF, hydrazine acetate, rt; CH2Cl2, TCA, DBU, -20~0 °C; CH2Cl2,
4 Å molecular sieves, Mu, BF3 Et2O, -40~-30 °C; 34%. g) MeOH, Na, rt, 97 %. h)
THF/H2O (4/1), Ph3P, silica gel, 50 °C, 99%.
3-Azido-3-deoxy-1,2:5,6-di-O-cyclohexylidene-D-glucofuranose (18): To a solution
of 17 [10] (3 g, 8.8 mmol) in dry CH2Cl2 (15 mL), stirred at 0 °C under N2, was added
Dess-Martin Periodinane (DMP) (5.61 g, 13.2 mmol, 1.5 equiv) in one portion. After 5
minutes, the cooling bath was removed and the reaction mixture was stirred overnight at
room temperature. After dilution with CH2Cl2 (100 mL) and saturated NaHCO3
containing Na2S2O3 (0.12 g/mL, 100 mL), the mixture was stirred vigorously until the two
layers were clear. After separation of the two layers, the organic phase was washed
with NaHCO3 containing Na2S2O3 (100 mL). The aqueous phase was extracted with

ACCEPTED MANUSCRIPT
17
EtOAc (100 mL), the combined organic phase was dried over MgSO4, filtered and
concentrated to afford a colorless syrup. The syrup was re-dissolved in ethanol (60 mL),
cooled to 0 °C and a solution of NaBH4 (0.43 g, 11.3 mmol, 1.3 equiv) in EtOH/H2O (1/1,
14 mL) was added dropwise over 35 minutes, and stirred under the same conditions for
a further 30 minutes. The cooling bath was then removed and the reaction mixture was
stirred for 1 h at room temperature, quenched with saturated NH4Cl, and water was
added until all solids were dissolved. The ethanol was then evaporated under vacuum.
The product was extracted with EtOAc (3 x 100 mL), washed with brine (150 mL), dried
over MgSO4, filtered and evaporated to dryness, then dried overnight under vacuum to
1
afford a white foam (2.91 g) of 1,2:5,6-di-O-cyclohexylidene-
D
-allofuranose [11]. H
NMR (d6-acetone, 400 MHz):
δ
5.76 (d, 1 H, J 4.0 Hz, H-1), 4.58 (t, 1 H, J2,3 4.0 Hz, H-
2), 4.28 (dt, 1 H, J5,6 2.8 Hz, H-5), 4.00 (dd, 1 H, J4,5, 6.8 Hz, J3,4 8.0 Hz, H-4), 3.93 (m, 3
H, H-3 & H-6), 2.84 (s, 1 H, OH), 1.80~1.30 (m, 20 H). 13C NMR (d6-acetone, 100 MHz):
δ
112.5, 109.5, 103.7, 79.3, 79.1, 75.4, 72.5, 64.5, 36.2, 36.0, 35.8, 35.1, 25.2, 24.9,
23.9, 23.86, 23.8, 23.6. ESI-HRMS: Calcd for [C18H28O6 + Na]+: 363.1784; Found m/z:
363.1783. The white foam was dissolved in dry CH2Cl2 (85 mL) and dry pyridine (12
mL), stirred at 0 °C under N2, the triflic anhydride (3.1 mL, 2.2 equiv) was added
dropwise. The reaction mixture was stirred for 2 h under the same conditions, diluted
with CH2Cl2 (100 mL), washed with cold 1 M HCl (100 mL), cold saturated NaHCO3
(100 mL) and cold brine (100 mL), dried over MgSO4, filtered and concentrated, dried
under vacuum to afford a yellow foam. To the yellow foam were added dry DMF (30 mL)
and NaN3 (4.7 g), and the suspension was stirred vigorously overnight at room
temperature under N2. The reaction mixture was diluted with EtOAc (300 mL), washed
with water (2 x 100 mL) and brine (2 x 100 mL), dried over MgSO4, filtered and
concentrated. The resulting residue was purified by flash column chromatography (15:1
1
petroleum ether-EtOAc) to afford 18 as a colorless syrup (3.09 g, 96%). H NMR (d6-
acetone, 400 MHz):
δ 5.86 (d, 1 H, J1,2 3.6 Hz, H-1), 4.68 (d, 1 H, H-2), 4.22 (m, 1 H, H-
5), 4.19 (d, 1 H, J3,4, 3.6 Hz, H-3), 4.11 (dd, 1 H, J5,6a 6.0 Hz, J6a,6b 8.4 Hz, H-6a), 4.10
13
(dd, 1 H, J4,5 8.8 Hz, H-4), 3.87 (dd, 1 H, J5,6b 5.2 Hz, H-6b), 1.70~1.30 (m, 20 H). C
NMR (d6-acetone, 100 MHz):
δ 112.4, 109.7, 104.9, 83.2, 80.7, 73.1, 67.1, 67.0, 36.6,

ACCEPTED MANUSCRIPT
18
36.3, 35.5, 34.7, 25.1, 24.8, 24.0, 23.9, 23.7, 23.6. ESI-HRMS: Calcd for [C18H27N3O5 +
Na]+: 388.1848; Found m/z: 388.1855.
3-Azido-3-deoxy-1,2,4,6-tetra-O-acetyl-D-glucopyranose (19): To compound 18 (2.98
g, 8.2 mmol) were added TFA (20 mL) and water (20 mL), and the mixture was stirred
for 2 days at room temperature. After evaporation and co-evaporation with toluene three
times, the resulting syrup was dried for 2 h under vacuum. To the residue were added
pyridine (50 mL) and acetic anhydride (25 mL), and the mixture was stirred overnight at
room temperature. After evaporation, water was added to the resulting residue and the
mixture was stirred for 0.5 h at room temperature and extracted with EtOAc (2 x 150
mL). The organic phase was washed with 1 M HCl (150 mL), saturated NaHCO3 (2 x
150 mL) and brine (150 mL), dried over MgSO4, filtered and concentrated. The resulting
residue was purified by flash column chromatography (3:1 & 7:3 petroleum ether-
EtOAc) to afford 19 as a colorless syrup (2.38 g, 78%; α/β = 1.2/1). ESI-MS: Calcd for
[C14H19N3O9 + Na]+: 396.1; Found m/z: 396.4.
3-Azido-3-deoxy-2,4,6-tri-O-acetyl-α-
general procedure A, the reaction mixture was stirred for 2 days at 0 °C, 20 was
obtained as a colorless syrup in 76% yield. 1H NMR (CDCl3, 400 MHz):
5.74 (dd, 1 H,
D-glucopyranosyl fluoride (20): See the
δ
J1,2 2.8 Hz, JH,F 53.2 Hz, H-1), 5.04 (t, 1 H, J3,4 = J4,5 10.2 Hz, H-4), 4.80 (dddd, 1 H, J2,3
10.8 Hz, JH,F 23.6 Hz, H-2), 4.24 (dd, 1 H, J5,6a 4.6 Hz, J6a,6b 12.6 Hz, H-6a), 4.13 (dd, 1
H, J5,6b 2.0 Hz, H-6b), 4.09 (m, 1 H, H-5), 3.99 (t, 1 H, H-3), 2.19 (s, 3 H, CH3CO), 2.14
(s, 3 H, CH3CO), 2.10 (s, 3 H, CH3CO). 13C NMR (CDCl3, 75 MHz):
169.2, 103.5 (d, JC,F 228.0 Hz), 71.0 (d, JC,F 24.0 Hz), 70.1 (d, JC,F 4.0 Hz), 67.3, 61.3,
δ 170.7, 169.9,
60.6, 20.8, 20.72, 20.7. 19F-NMR (CDCl3, 282 MHz):
[C12H16N3FO7 + Na]+: 356.0870; Found m/z: 356.0873.
δ -151.0. ESI-HRMS: Calcd for
3-Azido-3-deoxy-α-D-glucopyranosyl fluoride (21): See general procedure B, the
mixture was stirred for 30 h at 0 °C. The resulting residue was purified by flash column
1
chromatography to afford 21 as a colorless syrup in 97% yield. H NMR (CD3OD, 400
MHz):
δ 5.46 (dd, 1 H, J1,2 2.4 Hz, JH,F 53.6 Hz, H-1), 3.73 (m, 1 H, H-5), 3.63 (m, 2 H,
H-6), 3.50 (t, 1 H, J2,3 = J3,4 9.8 Hz, H-3), 3.45~3.31 (m, 2 H, H-2 & H-4). 13C NMR

ACCEPTED MANUSCRIPT
19
(CD3OD, 100 MHz):
δ
109.5 (d, JC,F 223.0 Hz), 77.1 (d, JC,F 3.0 Hz), 73.0 (d, JC,F 25.0
Hz), 70.3, 69.0, 62.9. 19F-NMR (CD3OD, 282 MHz):
[C6H10N3FO4 - H]-: 206.0577; Found m/z: 206.0581.
δ -152.7. ESI-HRMS: Calcd for
3-Amino-3-deoxy-α-
was obtained as a foam in 96% yield. 1H NMR (D2O, 400 MHz):
D
-glucopyranosyl fluoride (22): See general procedure C, 22
5.56 (dd, 1 H, J1,2 2.8
δ
Hz, JH,F 53.6 Hz, H-1), 3.78 (dd, 1 H, J5,6a 1.6 Hz, J6a,6b 11.6 Hz, H-6a), 3.73 (m, 1 H, H-
5), 3.68 (dd, 1 H, J5,6b 5.2 Hz, H-6b), 3.43 (dddd, 1 H, J2,3 10.4 Hz, JH,F 26.4 Hz, H-2),
3.31 (t, J3,4 = J4,5 9.2 Hz, H-4), 2.96 (t, 1 H, H-3). 13C NMR (CD3OD, 100 MHz):
(d, JC,F 223.0 Hz), 74.9 (d, JC,F 3.0 Hz), 71.7 (d, JC,F 24.0 Hz), 68.9, 60.9, 54.6. 19F-
δ 107.0
NMR (D2O, 282 MHz):
Found m/z: 182.0827.
δ
-150.8. ESI-HRMS: Calcd for [C6H12NFO4 + H]+: 182.0829;
4-Methylumbelliferyl 3-azido-3-deoxy-2,4,6-tri-O-acetyl-β-
See general procedure D, eluent (2:1 petroleum ether-EtOAc), 3-azido-3-deoxy-2,4,6-
tri-O-acetyl- -glucopyranose was obtained as a colorless syrup in 71% yield. See
general procedure E, eluent (2:1 & 3:2 petroleum ether-EtOAc), 23 was obtained as a
white solid in 48% yield (overall yield: 34%). 1H NMR (CDCl3, 400 MHz):
7.52 (d, 1 H,
D-glucopyranoside (23):
D
δ
J 8.4 Hz, Ar-H), 6.91 (m, 2 H, Ar-H), 6.19 (d, 1 H, J 0.8 Hz, Ar-H), 5.22 (dd, 1 H, J2,3
10.0 Hz, H-2), 5.20 (d, 1 H, J1,2 7.6 Hz, H-1), 5.06 (t, 1 H, J3,4 = J4,5 10.0 Hz, H-4), 4.25
(dd, 1 H, J5,6a 6.0 Hz, J6a,6b 12.4 Hz, H-6a), 4.17 (dd, 1 H, J5,6b 2.4 Hz, H-6b), 3.88
(dddd, 1 H, H-5), 3.77 (t, 1 H, H-3), 2.41 (d, J 1.2 Hz, 3 H, Ar-CH3), 2.16 (s, 3 H,
13
CH3CO), 2.15 (s, 3 H, CH3CO), 2.12 (s, 3 H, CH3CO). C NMR (CDCl3, 100 MHz):
δ
170.8, 169.3, 169.1, 160.9, 159.3, 155.0, 152.4, 125.9, 115.7, 114.3, 113.1, 103.9, 98.7,
73.4, 70.8, 68.4, 64.2, 62.0, 20.84, 20.79, 20.76, 18.8. ESI-HRMS: Calcd for
[C22H23N3O10 + Na]+: 512.1281; Found m/z: 512.1281.
4-Methylumbelliferyl 3-azido-3-deoxy-β-
D
-glucopyranoside (24): See general
1
procedure F, 24 was obtained as a white solid in 97% yield. H NMR (d6-DMSO + D2O,
400 MHz):
(d, 1 H, J1,2 7.6 Hz, H-1), 3.70~3.23 (m, 6 H, remaining protons of sugar ring), 2.31 (s, 3
H, Ar-CH3). 13C NMR (d6-DMSO + D2O, 100 MHz):
159.8, 159.4, 153.9, 153.0, 126.1,
δ 7.71 (d, 1 H, J 9.4 Hz, Ar-H), 7.04 (m, 2 H, Ar-H), 6.24 (s, 1 H, Ar-H), 5.15
δ

ACCEPTED MANUSCRIPT
20
113.9, 113.0, 111.4, 102.8, 99.2, 77.0, 70.9, 69.0, 67.5, 59.6, 17.7. ESI-HRMS: Calcd
for [C16H17N3O7 + Na]+: 386.0963; Found m/z: 386.0964.
4-Methylumbelliferyl 3-amino-3-deoxy-β-
D
-glucopyranoside (25): See general
1
procedure G, 25 was obtained as a white solid in 99% yield. H NMR (d6-DMSO + D2O,
400 MHz):
δ 7.70 (d, 1 H, J 9.2 Hz, Ar-H), 7.04 (m, 2 H, Ar-H), 6.24 (s, 1 H, Ar-H), 5.07
(d, 1 H, J1,2 7.6 Hz, H-1), 4.02 (d, 1 H, J6a,6b 7.6 Hz, H-6a), 3.45 (m, 2 H, H-5 & H-6b),
3.25 (dd, 1 H, H-2), 3.15 (t, 1 H, J4,5 9.0 Hz, H-4), 2.72 (t, 1 H, J2,3 = J3,4 9.6 Hz, H-3),
2.40 (s, 3 H, Ar-CH3). 13C NMR (d6-DMSO + D2O, 100 MHz):
δ 160.8, 160.7, 155.0,
154.0, 127.1, 114.7, 114.0, 112.3, 103.8, 100.8, 78.7, 72.8, 69.5, 61.0, 59.3, 18.7. ESI-
HRMS: Calcd for [C16H19NO7 + Na]+: 360.1053; Found m/z: 360.1059.
Scheme 4. Reagents and conditions: a). CH2Cl2, TEA, 1-(acetoxy)-benzotriazole, rt,
79%. b) CH2Cl2, pyridine, Tf2O, 0 °C; DMF, NaN3, rt; 91%. c) CH2Cl2, Ac2O,
HF/pyridine, 0 °C, 86%. d) MeOH, NH3, 0 °C, 98%. e) MeOH, Pd/C, H2, rt, 98%. f) DMF,

ACCEPTED MANUSCRIPT
21
hydrazine acetate, rt; CH2Cl2, TCA, DBU, -20~0 °C; DCM, 4 Å molecular sieves, Mu,
BF3 Et2O, -40~-30 °C; 32%. g) MeOH, Na, rt, 92%. h) THF/H2O (4/1), Ph3P, silica gel,
50 °C, 90%.
1,2,3,6-Tetra-O-acetyl-D-glucopyranose (26): To a solution of 9 (4.7 g, 15.4 mmol) in
anhydrous CH2Cl2 (100 mL) and Et3N (3.2 mL) was added 1-(acetoxy)-benzotriazole
[29] (3.26 g, 18.4 mmol, 1.2 equiv) in one portion [30, 31], and the mixture was stirred
for 5 h at room temperature under N2. After completion, the mixture was diluted with
CH2Cl2 (50 mL), washed with brine (2 x 100 mL), dried over MgSO4, filtered and
concentrated. The resulting residue was purified by flash column chromatography
(55:45, petroleum ether-EtOAc) to afford 26 as a syrup (4.27 g, 79%; α/β = 1/1.3). ESI-
MS: Calcd for [C14H20O10 + Na]+: 371.1; Found m/z: 371.2.
4-Azido-4-deoxy-1,2,3,6-tetra-O-acetyl-D-galactopyranose (27): To a solution of 26
(4.23 g, 12.2 mmol) in dry CH2Cl2 (120 mL) and dry pyridine (20 mL), stirred at 0 °C
under N2, was added triflic anhydride (4.4 mL, 26.2 mmol, 2.1 equiv) dropwise, and the
mixture stirred for 1 h under the same conditions. After completion, the reaction mixture
was diluted with CH2Cl2 (150 mL), washed with cold 1 M HCl (2 x 200 mL), cold
saturated NaHCO3 (200 mL) and cold brine (200 mL), dried over MgSO4, filtered and
concentrated to afford a brownish foam. To the brownish foam were added dry DMF (50
mL) and NaN3 (6.88 g, 105.8 mmol, 8.7 equiv), and the reaction mixture stirred
vigorously for 1.5 h at room temperature under N2. The reaction mixture was diluted with
EtOAc (500 mL), washed with water (2 x 250 mL) and brine (250 mL), dried over
MgSO4, filtered and concentrated. The resulting residue was purified by flash column
chromatography (65:35, petroleum ether-EtOAc) to afford 27 as a syrup (91%; α/β =
1/1.23). ESI-MS: Calcd for [C14H19N3O9 + Na]+: 396.1; Found m/z: 396.2.
4-Azido-4-deoxy-2,3,6-tri-O-acetyl-α-D-galactopyranosyl fluoride (28): See the
general procedure A, the reaction mixture was stirred overnight at 0 °C, 28 was
1
obtained as a colorless syrup in 86%. H NMR (CDCl3, 400 MHz):
δ 5.79 (dd, 1 H, J1,2
2.4 Hz, JH,F 53.2 Hz, H-1), 5.48 (dd, 1 H, J3,4 3.2 Hz, H-3), 5.31 (dddd, 1 H, J2,3 10.8 Hz,
JH,F 23.6 Hz, H-2), 4.36~4.25 (m, 4 H, H-4,5,6), 2.21 (s, 3 H, CH3CO), 2.18 (s, 3 H,

ACCEPTED MANUSCRIPT
22
CH3CO), 2.16 (s, 3 H, CH3CO). 13C NMR (CDCl3, 75 MHz):
δ 170.5, 170.2 (2 C), 104.3
(d, JC,F 227.6 Hz), 69.3, 68.7 (d, JC,F 3.5 Hz), 67.6 (d, JC,F 23.7 Hz), 62.6, 60.3, 20.9,
20.7, 20.6. 19F-NMR (CDCl3, 282 MHz):
Na]+: 356.0870; Found m/z: 356.0860.
δ -151.6. ESI-HRMS: Calcd for [C12H16N3FO7 +
4-Azido-4-deoxy-α-D-galactopyranosyl fluoride (29): See general procedure B, the
mixture was stirred overnight at 0 °C. The resulting residue was purified by flash column
1
chromatography to afford 29 as a colorless syrup in 98% yield. H NMR (CD3OD, 400
MHz):
(dddd, 1 H, J2,3 9.2 Hz, JH,F 26.0 Hz, H-2), 3.68(d, 2 H, J5,6 6.4 Hz, H-6). 13C NMR
(CD3OD, 100 MHz): 107.7 (d, JC,F 223.6 Hz), 71.7 (d, JC,F 2.7 Hz), 69.9, 68.7 (d, JC,F
δ 5.55 (dd, 1 H, J1,2 2.8 Hz, JH,F 54.0 Hz, H-1), 4.08~4.03 (m, 3 H, H-3,4,5), 3.79
δ
24.2 Hz), 63.0, 60.9. 19F-NMR (CD3OD, 282 MHz):
[C6H10N3FO4 - H]-: 206.0577; Found m/z: 206.0579.
δ -153.5. ESI-HRMS: Calcd for
4-Amino-4-deoxy-α-
D
-galactopyranosyl fluoride (30): See general procedure C, 30
1
was obtained as a white solid in 98% yield. H NMR (CD3OD, 400 MHz):
δ 5.60 (dd, 1
H, J1,2 3.2 Hz, JH,F 54.0 Hz, H-1), 4.08 (m, 1 H, H-5), 3.90 (dd, 1 H, J5,6a 4.4 Hz, J6a,6b
13
10.4 Hz H-6a), 3.80~3.69 (m, 3 H, H-2, 3, 6b), 3.38 (brs, 1 H, H-4). C NMR (CD3OD,
100 MHz):
δ 107.7 (d, JC,F 224.0 Hz), 71.8 (d, JC,F 3.0 Hz), 68.5, 68.2 (d, JC,F 24.0 Hz),
61.1, 52.5. 19F-NMR (D2O, 282 MHz):
δ
-151.9. ESI-HRMS: Calcd for [C6H12NFO4 + H]+:
182.0829; Found m/z: 182.0822.
4-Methylumbelliferyl 4-azido-4-deoxy-2,3,6-tri-O-acetyl-β-
See general procedure D, eluent (2:1 petroleum ether-EtOAc), 4-azido-4-deoxy-2,3,6-
tri-O-acetyl- -galactopyranose was obtained as a colorless syrup in 73% yield. See
general procedure E, eluent (2:1 & 3:2 petroleum ether-EtOAc), 23 was obtained as a
white solid in 43% yield (overall yield: 31%). 1H NMR (CDCl3, 400 MHz):
7.55 (d, 1 H,
D-galactopyranoside (31):
D
δ
J 9.1 Hz, Ar-H), 6.95 (m, 2 H, Ar-H), 6.22 (d, 1 H, J 0.9 Hz, Ar-H), 5.60 (dd, 1 H, J2,3
10.4 Hz, H-2), 5.25 (dd, 1 H, J3,4 3.7 Hz, H-3), 5.10 (d, 1 H, J1,2 7.9 Hz, H-1), 4.38 (dd, 1
H, J5,6a 7.0 Hz, J6a,6b 11.6 Hz, H-6a), 4.29 (dd, 1 H, J5,6b 5.8 Hz, H-6b), 4.18 (brd, 1 H, H-
4), 4.03 (m, 1 H, H-5), 2.44 (d, J 0.9 Hz, 3 H, Ar-CH3), 2.20 (s, 3 H, CH3CO), 2.19 (s, 3
H, CH3CO), 2.12 (s, 3 H, CH3CO). 13C NMR (d6-acetone, 100 MHz):
δ 170.0, 169.4,

ACCEPTED MANUSCRIPT
23
169.2, 159.9, 159.6, 155.0, 152.7, 126.4, 115.3, 113.6, 112.7, 103.8, 98.4, 72.6, 71.3,
68.8, 62.7, 60.4, 20.0, 19.9, 19.7, 17.8. ESI-HRMS: Calcd for [C22H23N3O10 + Na]+:
512.1281; Found m/z: 512.1296.
4-Methylumbelliferyl 4-azido-4-deoxy-β-
D
-galactopyranoside (32): See general
1
procedure F, 32 was obtained as a white solid in 92% yield. H NMR (d6-DMSO + D2O,
400 MHz):
δ 7.68 (d, 1 H, J 9.6 Hz, Ar-H), 7.01 (m, 2 H, Ar-H), 6.22 (d, 1 H, J 0.8 Hz, Ar-
H), 5.03 (d, 1 H, J1,2 7.6 Hz, H-1), 3.93 (brd, 1 H, H-4), 3.84 (m, 1 H, H-5), 3.80 (dd, 1 H,
J2,3 9.6 Hz, J3,4 4.0 Hz, H-3), 3.54 (m, 1 H, H-2), 3.45 (m, 2 H, H-6), 2.38 (s, 3 H, Ar-
CH3). 13C NMR (d6-DMSO + D2O, 100 MHz):
δ 159.8, 159.4, 153.9, 153.0, 126.0, 113.7,
113.0, 111.3, 102.6, 99.6, 72.7, 72.4, 69.8, 61.8, 59.6, 17.7. ESI-HRMS: Calcd for
[C16H17N3O7 + Na]+: 386.0964; Found m/z: 386.0961.
4-Methylumbelliferyl 4-amino-4-deoxy-β-
D
-galactopyranoside (33): See general
1
procedure G, 33 was obtained as a white solid in 90% yield. H NMR (d6-DMSO + D2O,
400 MHz):
δ 7.70 (d, 1 H, J 9.6 Hz, Ar-H), 7.04 (m, 2 H, Ar-H), 6.24 (s, 1 H, Ar-H), 4.99
(d, 1 H, J1,2 7.2 Hz, H-1), 3.78 (m, 1 H, H-5), 3.70~3.30 (m, 4 H, H-2, H-3 & H-6), 3.03
13
(brs, 1 H, H-4), 2.40 (s, 3 H, Ar-CH3). C NMR (d6-DMSO + D2O, 100 MHz):
δ 160.8,
160.7, 155.0, 154.1, 127.1, 114.7, 114.0, 112.3, 103.7, 101.3, 75.5, 72.8, 70.6, 61.1,
52.3, 18.7. ESI-HRMS: Calcd for [C16H19NO7 + H]+: 338.1240; Found m/z: 338.1238.

ACCEPTED MANUSCRIPT
24
AcO
HO
HO
N3
N3
NH2
O
d
e
O
O
AcO
HO
HO
AcO
HO
HO
39
F
F
F
38
37
c
O
O
N3
AcO
AcO
OH
N3
O
O
O
b
a
O
O
OAc
O
OAc
O
O
O
36
35
34
f
AcO
HO
N3
N3
O
O
g
O
O
O
O
O
O
AcO
HO
40
OAc
41
OH
h
HO
NH2
O
O
O
O
HO
42
OH
Scheme 5. Reagents and conditions: a) CH2Cl2, pyridine, Tf2O, 0 °C; DMF, NaN3, rt;
75%. b) AcOH/H2O (2/1), 80 °C; pyridine, Ac2O, rt; 92%. c) CH2Cl2, HF/pyridine, 0 °C;
78%. d) MeOH, NH3, 0 °C; 95%. e) MeOH, Pd/C, H2, rt, 96%. f) CH2Cl2, EtOAc, Ac2O,
TiBr4, rt; CH2Cl2, 1 M NaOH, TBAB, Mu, rt; 58%. g) MeOH, Na, rt; 89%. h) THF/H2O
(4/1), Ph3P, silica gel, 50 °C, 94%.
6-Azido-6-deoxy-1,2,3,4-di-O-isopropylidene-α-
D
-galactopyranose (35): To
a
solution of 34 (0.91 g, 3.5 mmol) in dry CH2Cl2 (15 mL) and dry pyridine (6 mL), stirred
at 0 °C under N2, was added triflic anhydride (1.2 mL, 7.1 mmol, 2 equiv) dropwise, and
the reaction mixture was stirred for 1 h under the same conditions. After completion, the
mixture was diluted with DCM (70 mL), washed with cold saturated NaHCO3 (40 mL)
and cold brine (40 mL), dried over MgSO4, filtered and evaporated, co-evaporated twice
with toluene, and dried under vacuum for 1 h to afford a brownish foam. To this foam
were added dry DMF (10 mL) and NaN3 (0.7 g, 10.8 mmol, 3 equiv), and the reaction
mixture stirred vigorously for 1.5 h at room temperature under N2. The reaction mixture

ACCEPTED MANUSCRIPT
25
was diluted with EtOAc (100 mL), washed with water (2 x 50 mL) and brine (50 mL),
dried over MgSO4, filtered and concentrated. The resulting residue was purified by flash
column chromatography (9:1, petroleum ether-EtOAc with 1% Et3N) to afford 35 as a
1
colorless syrup (748 mg, 75%). H NMR (CDCl3, 400 MHz):
δ 5.53 (d, 1 H, J1,2 5.2 Hz,
H-1), 4.62 (dd, 1 H, J3,4 7.6 Hz, H-3), 4.32 (dd, 1 H, J2,3 2.4 Hz, H-2), 4.18 (dd, 1 H, J4,5
2.0 Hz, H-4), 3.90 (m, 1 H, H-5), 3.50 (dd, 1 H, J5,6a 8.0 Hz, J6a,6b 12.8 Hz, H-6a), 3.35
(dd, 1 H, J5,6b 5.4 Hz, H-6b), 1.54 (s, 3 H, CH3C), 1.45 (s, 3 H, CH3C), 1.333 (s, 3 H,
CH3C), 1.327 (s, 3 H, CH3C). 13C NMR (CDCl3, 75 MHz):
δ 109.7, 108.9, 96.5, 71.3,
70.9, 70.5, 67.1, 50.8, 26.14, 26.05, 25.0, 24.5. ESI-HRMS: Calcd for [C12H19N3O5 +
Na]+: 308.1222; Found m/z: 308.1227.
6-Azido-6-deoxy-1,2,3,4-tetra-O-acetyl-D-galactopyranose (36): A mixture of 35 (9.05
g, 31.8 mmol), AcOH (120 mL) and H2O (60 mL) was stirred for 27 h at 80 °C,
evaporated and co-evaporated with toluene twice, then dried under vacuum for 2 h to
afford a syrup. To this syrup were added pyridine (120 mL) and Ac2O (60 mL), and the
mixture stirred overnight at room temperature. After evaporation, ice-water was added
and the mixture was stirred for 0.5 h, extracted with EtOAc (2 x 250 mL). The organic
phase was washed with 1 M HCl (2 x 250 mL), water (250 mL), saturated NaHCO3 (2 x
250 mL) and brine (250 mL), dried over MgSO4, filtered and concentrated. The resulting
residue was purified by flash column chromatography (3:1, petroleum ether-EtOAc) to
afford 36 as a syrup (10.84 g, 92%; α/β=1/1.38). ESI-MS: Calcd for [C14H19N3O9 + Na]+:
396.1; Found m/z: 396.0.
6-Azido-6-deoxy-2,3,4-tri-O-acetyl-α-
general procedure A, the reaction mixture was stirred overnight at 0 °C, 37 was
obtained as a colorless syrup in 78% yield. 1H NMR (CDCl3, 400 MHz):
5.82 (dd, 1 H,
D-galactopyranosyl fluoride (37): See the
δ
J1,2 2.8 Hz, JH,F 53.2 Hz, H-1), 5.50 (brd, 1 H, H-4), 5.36 (dd, 1 H, J3,4 2.8 Hz, H-3), 5.19
(dddd, 1 H, J2,3 10.8 Hz, JH,F 24.0 Hz, H-2), 4.31 (m, 1 H, H-5), 3.49 (dd, 1 H, J5,6a 7.2
Hz, J6a,6b 12.8 Hz, H-6a), 3.26 (dd, 1 H, J5,6b 5.2 Hz, H-6b), 2.18 (s, 3 H, CH3CO), 2.13
(s, 3 H, CH3CO), 2.05 (s, 3 H, CH3CO). 13C NMR (CDCl3, 75 MHz):
169.9, 104.3 (d, JC,F 227.0 Hz), 70.2 (d, JC,F 3.0 Hz), 68.1, 67.4 (d, JC,F 24.0 Hz), 67.1,
50.5, 20.8, 20.7 (2 C). 19F-NMR (CDCl3, 282 MHz):
-150.6. ESI-HRMS: Calcd for
δ 170.4, 170.1,
δ

ACCEPTED MANUSCRIPT
26
[C12H16N3FO7 + Na]+: 356.0870; Found m/z: 356.0869.
6-Azido-6-deoxy-α-D-galactopyranosyl fluoride (38): See general procedure B, the
mixture was stirred overnight at 0 °C. The resulting residue was purified by flash column
1
chromatography to afford 38 as a colorless syrup in 95% yield. H NMR (D2O, 400
MHz):
δ 5.62 (dd, 1 H, J1,2 2.8 Hz, JH,F 53.2 Hz, H-1), 4.10 (dd, 1 H, H-5), 3.93 (brd, 1 H,
J3,4 2.0 Hz, H-4), 3.83~3.70 (m, 2 H, J2,3 10.4 Hz, JH,F 25.2 Hz, H-2 & H-3), 3.52 (dd, 1
13
H, J5,6a 8.8 Hz, J6a,6b 12.8 Hz, H-6a), 3.41 (dd, 1 H, J5,6b 4.8 Hz, H-6b). C NMR (D2O,
100 MHz):
δ
107.5 (d, JC,F 223.0 Hz), 71.9 (d, JC,F 3.0 Hz), 69.1, 68.8, 67.8 (d, JC,F 24.0
19
Hz), 50.9. F-NMR (CD3OD, 282 MHz):
H]-: 206.0577; Found m/z: 206.0584.
δ -152.3. ESI-HRMS: Calcd for [C6H10N3FO4 -
6-Amino-6-deoxy-α-
D
-galactopyranosyl fluoride (39): See general procedure C, 39
1
was obtained as a white solid in 96% yield. H NMR (D2O, 400 MHz):
δ 5.61 (dd, 1 H,
J1,2 2.4 Hz, JH,F 53.6 Hz, H-1), 3.98 (dd, 1 H, H-5), 3.93 (brd, 1 H, J3,4 2.8 Hz, H-4),
3.82~3.69 (m, 2 H, J2,3 10.0 Hz, JH,F 25.0 Hz, H-2 & H-3), 2.92 (dd, 1 H, J5,6a 8.4 Hz,
J6a,6b 13.6 Hz, H-6a), 2.85 (dd, 1 H, J5,6b 3.6 Hz, H-6b). 13C NMR (D2O, 100 MHz):
δ
19
107.5 (d, JC,F 222.0 Hz), 72.5, 69.4, 69.0, 67.9 (d, JC,F 25.0 Hz), 40.8. F-NMR (D2O,
282 MHz):
182.0827.
δ
-152.3. ESI-HRMS: Calcd for [C6H12NFO4 + H]+: 182.0829; Found m/z:
4-Methylumbelliferyl 6-azido-6-deoxy-2,3,4-tri-O-acetyl-β-D-galactopyranoside (40):
To a solution of 36 (1.14 g, 3.1 mmol) in dry CH2Cl2 (17 mL), dry EtOAc (1.7 mL) and
Ac2O (0.3 mL), stirred at room temperature under N2, was added TiBr4 (2.25 g, 6.1
mmol, 2 equiv), and the reaction mixture stirred for 2 days under the same conditions
[32]. An additional 10% of TiBr4 was added each day until completion. The reaction
mixture was diluted with CH2Cl2 (125 mL), washed with cold water (2 x 80 mL), cold
brine (80 mL), dried over MgSO4, filtered and concentrated, and dried under vacuum for
2 h to afford a foam. To the foam were added CH2Cl2 (16 mL), 1 M NaOH (8 mL), Mu
(1.1 g, 6.2 mmol, 2 equiv) and tetra-n-butylammonium bromide (TBAB, 1 g), and the
reaction mixture stirred vigorously overnight at room temperature [33]. The reaction
mixture was diluted with CH2Cl2 (125 mL), washed with 1 M NaOH (2 x 60 mL),

ACCEPTED MANUSCRIPT
27
saturated NaHCO3 (60 mL) and brine (60 mL), dried over MgSO4, filtered and
concentrated. The resulting residue was purified by flash column chromatography (2:1
1
petroleum ether-EtOAc) to afford 40 as a white solid (0.86 g, 58%). H NMR (CDCl3,
400 MHz):
δ 7.54 (d, 1 H, J 9.2 Hz, Ar-H), 6.98 (m, 2 H, Ar-H), 6.20 (d, 1 H, J 1.2 Hz, Ar-
H), 5.50 (dd, 1 H, J2,3 10.8 Hz, H-2), 5.45 (brd, 1 H, H-4), 5.17 (d, 1 H, J1,2 7.6 Hz, H-1),
5.14 (dd, 1 H, J3,4 3.2 Hz, H-3), 3.99 (m, 1 H, H-5), 3.58 (dd, 1 H, J5,6a 8.0 Hz, J6a,6b 13.2
Hz, H-6a), 3.28 (dd, 1 H, J5,6b 4.8 Hz, H-6b), 2.41 (d, J 1.2 Hz, 3 H, Ar-CH3), 2.22 (s, 3
H, CH3CO), 2.08 (s, 3 H, CH3CO), 2.03 (s, 3 H, CH3CO). 13C NMR (CDCl3, 100 MHz):
δ
170.3, 170.1, 169.5, 160.0, 159.3, 155.0, 152.3, 126.0, 115.8, 113.7, 113.4, 104.6, 99.2,
73.3, 70.7, 68.4, 67.7, 50.7, 20.84, 20.8, 20.7, 18.8. ESI-HRMS: Calcd for [C22H23N3O10
+ Na]+: 512.1281; Found m/z: 512.1277.
4-Methylumbelliferyl 6-azido-6-deoxy-β-
D
-galactopyranoside (41): See general
1
procedure F, 41 was obtained as a white solid in 89% yield. H NMR (d6-DMSO + D2O,
400 MHz):
δ 7.70 (d, 1 H, J 8.8 Hz, Ar-H), 7.03 (m, 2 H, Ar-H), 6.23 (d, 1 H, J 0.8 Hz,
Ar-H), 5.08 (d, 1 H, J1,2 8.0 Hz, H-1), 3.94 (m, 1 H, H-5), 3.65 (brd, 1 H, H-4), 3.61 (dd, 1
H, J2,3 9.6 Hz, H-2), 3.54 (m, 1 H, H-6a), 3.47 (dd, 1 H, J3,4 3.2 Hz, H-3), 3.26 (dd, 1 H,
J5,6b 3.6 Hz, J6a,6b 12.8 Hz, H-6b), 2.39 (d, 3 H, J 0.8 Hz, Ar-CH3). 13C NMR (d6-DMSO +
D2O, 100 MHz):
δ 160.8, 160.5, 155.0, 154.1, 127.0, 114.7, 114.0, 112.2, 103.6, 100.7,
74.7, 73.1, 70.2, 69.2, 51.7, 18.7. ESI-HRMS: Calcd for [C16H17N3O7 + H]+: 364.1145;
Found m/z: 364.1146.
4-Methylumbelliferyl 6-amino-6-deoxy-β-
D
-galactopyranoside (42): See general
1
procedure G, 42 was obtained as a white solid in 94% yield. H NMR (d6-DMSO + D2O,
400 MHz):
δ 7.69 (d, 1 H, J 9.6 Hz, Ar-H), 7.05 (m, 2 H, Ar-H), 6.23 (s, 1 H, Ar-H), 4.97
(d, 1 H, J1,2 8.0 Hz, H-1), 3.73~3.57 (m, 3 H, H-2,4 & 5), 3.43 (dd, 1 H, J2,3 9.6 Hz, J3,4
13
3.2 Hz, H-3), 2.81 (d, 2 H, J5,6 6.0 Hz, H-6), 2.39 (d, 3 H, J 0.4 Hz, Ar-CH3). C NMR
(d6-DMSO + D2O, 100 MHz):
δ 160.8, 160.7, 155.1, 153.9, 127.0, 114.6, 113.9, 112.3,
103.6, 101.1, 75.9, 73.8, 70.7, 69.2, 42.1, 18.7. ESI-HRMS: Calcd for [C16H19NO7 + H]+:
338.1240; Found m/z: 338.1238.

ACCEPTED MANUSCRIPT
28
Scheme 6. Reagents and conditions: a) pyridine, BzCl, 0 °C; CH2Cl2, pyridine, Tf2O, 0
°C; DMF, NaN3, rt; MeOH, NaOMe, rt; pyridine, Ac2O, rt; 33%. b) CH2Cl2, Ac2O,
HF/pyridine, 0 °C, 87%; c) MeOH, NH3, 0 °C, 99%. d) MeOH, Pd/C, H2, rt. e) DMF,
hydrazine acetate, rt; CH2Cl2, TCA, DBU, -20 0 °C; CH2Cl2, Mu, BF3 Et2O, -40 -20 °C,
4Å molecular sieves; 41%. f) CH2Cl2, MeOH, Na, rt, 98%. g) THF/H2O (4/1), Ph3P, silica
gel, 50 °C, 94%
4-Azido-4-deoxy-2,3-di-O-acetyl-α-
D
-xylosyl fluoride (45): See the general procedure
-xylose [34, 35] in dry CH2Cl2 and
A, a solution of 4-azido-4-deoxy-1,2,3-tri-O-acetyl-
D
HF/Pyridine was stirred for 24 h at 0 °C. After flash column chromatography (9:1
1
petroleum ether-EtOAc), 45 was obtained as a white solid in 87% yield. H NMR
(CDCl3, 400 MHz):
δ 5.66 (dd, 1 H, J1,2 2.8 Hz, JH,F 52.8 Hz, H-1), 5.41 (t, 1 H, J3,4 9.6
Hz, H-3), 4.87 (dddd, 1 H, J2,3 10.0 Hz, JH,F 24.0 Hz, H-2), 3.92 (dd, 1 H, J4,5a 2.4 Hz,
J5a,5b 5.6 Hz, H-5a), 3.76~3.73 (m, 2 H, H-4 & H-5b), 2.12 (s, 3 H, CH3CO), 2.10 (s, 3 H,
13
CH3CO). C NMR (CDCl3, 75 MHz):
δ 170.3, 169.8, 104.3 (d, JC,F 228.0 Hz), 70.6 (d,
JC,F 24.0 Hz), 70.1, 61.5 (d, JC,F 4.0 Hz), 58.8, 20.8, 20.7. 19F-NMR (CDCl3, 282 MHz):
δ
-150.9. ESI-HRMS: Calcd for [C9H12N3FO5 + Na]+: 284.0659; Found m/z: 284.0655.