Stereospecific synthesis of methyl 2-amino-2-deoxy-(6S)-deuterio-α,β-D-glucopyranoside and methyl 2,6-diamino-2,6-dideoxy-(6R)-deuterio-α,β-D-glucopyranoside: Side chain conformations of the 2-amino-2-deoxy and 2,6-diamino-2,6-dideoxyglucopyranosides

Article abstract of DOI:10.1016/j.carres.2017.05.015

The stereospecifically labeled 6-monodeuterio methyl 2,6-diamino-2,6-dideoxy-α- and β- D-glucopyranosides were synthesized with a view to determining their side chain conformations. NMR studies in D₂O at pH 5 and pH 11 reveal both anomers to adopt very predominantly the gt conformation consistent with the gauche conformation of 2-aminoethanol and its acetate salt. In contrast, as also revealed with the help of stereospecifically-labelled monodeuterio isotopomers, the methyl 2-amino-2-deoxy-α- and β- D-glucopyranosides are an approximately 1:1 mixture of gg and gt conformers as is found in glucopyranose itself.

Full text of DOI:10.1016/j.carres.2017.05.015

Carbohydrate Research 448 (2017) 10e17
Contents lists available at ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Stereospecific synthesis of methyl 2-amino-2-deoxy-(6S)-deuterio-
a,
b-D
-glucopyranoside and methyl 2,6-diamino-2,6-dideoxy-(6R)-
deuterio- -glucopyranoside: Side chain conformations of the 2-
amino-2-deoxy and 2,6-diamino-2,6-dideoxyglucopyranosides
a
,b-
D
,
, **
Takayuki Kato a *, Andrea Vasella b, David Crich a
a Department of Chemistry, Wayne State University, 5101 Cass Avenue Detroit, MI 48202, USA
b Organic Chemistry Laboratory, ETH Zurich, 8093 Zurich, Switzerland
a r t i c l e i n f o
a b s t r a c t
Article history:
The stereospecifically labeled 6-monodeuterio methyl 2,6-diamino-2,6-dideoxy-a- and b- D-glucopyr-
Received 30 March 2017
Received in revised form
15 May 2017
Accepted 15 May 2017
Available online 19 May 2017
anosides were synthesized with a view to determining their side chain conformations. NMR studies in
D2O at pH 5 and pH 11 reveal both anomers to adopt very predominantly the gt conformation consistent
with the gauche conformation of 2-aminoethanol and its acetate salt. In contrast, as also revealed with
the help of stereospecifically-labelled monodeuterio isotopomers, the methyl 2-amino-2-deoxy-
-glucopyranosides are an approximately 1:1 mixture of gg and gt conformers as is found in gluco-
pyranose itself.
a- and
b- D
Keywords:
© 2017 Elsevier Ltd. All rights reserved.
Side chain conformation
Deuterium labelling
Conformational analysis
Aminosugars
1. Introduction
solution (pH < 1 and 4.75), and as a 70:30 mixture of gg, and gt
conformers in aqueous base (pH > 12) [1]. However, in the absence
of the crucial rigorous assignment of the H6R and H6S resonances
the coupling constants are also consistent with the tg conformer
being the most populated state of 1 [1]. This ambiguity precludes
the use of 1 as model for the side chain conformation of the 2,6-
diamino-2,6-dideoxyglucopyranosides and prompted the synthe-
sis and conformational analysis of both anomers, 2 and 3, of methyl
Ongoing projects in our laboratories necessitated knowledge of
the side chain conformation of the 2-amino-2-deoxy- and espe-
cially the 2,6-diamino-2,6-dideoxyglucopyranosides. On the basis
of numerous X-ray crystallographic and NMR spectroscopic studies
the hydroxymethyl side chain of glucopyranose and its glycosides is
considered to populate an approximately 60:40 equilibrium
mixture of the staggered gg and gt conformations; the tg conformer
being destabilized by dipolar repulsions between the C6-O6 and
C4-O4 bonds (Fig. 1) [1e3].
2,6-diamino-2,6-dideoxy-
D
-glucopyranoside
stereospecifically
deuterium labeled at the pro-6R site that we report here. Also re-
ported are the syntheses and conformational analysis of both
Analysis of the H5-H6R and H5-H6S coupling constants of ~9.2
and 3.1 Hz, respectively, led Bock and Duus to conclude that the side
anomers, 4 and 5, of 6S-monodeuterio-2-amino-2-deoxy-D-gluco-
pyranoside.
chain of methyl 6-amino-6-deoxy-b-D-glucopyranoside 1 exists as
an 80:16:4 mixture of the gg, gt, and tg conformers in acidic
* Corresponding author.
** Corresponding author.
E-mail addresses: tkato@chem.wayne.edu (T. Kato), dcrich@chem.wayne.edu (D. Crich).
http://dx.doi.org/10.1016/j.carres.2017.05.015
0008-6215/© 2017 Elsevier Ltd. All rights reserved.

T. Kato et al. / Carbohydrate Research 448 (2017) 10e17
11
anhydroglucose 8 in 82% yield (Scheme 1). Zemplen deacetylation
then gave a triol which on reaction with toluenesulfonyl chloride in
pyridine gave the 2,4-di-O-tosylate 9 in 82% yield for the two steps.
Subsequent exposure to sodium methoxide in methanol afforded
77% of the galacto-configured epoxide 10 that on treatment with
benzyl alcohol under Lewis acidic conditions gave the 4-O-benzyl
derivative 11 in 78% yield. Treatment of 11 with sodium methoxide
resulted in the manno-epoxide 12 in 88% yield, which on heating
with sodium azide in hot aqueous DMF, followed by benzylation of
the intermediate alcohol gave the 1,6-anhydroglucosamine deriv-
ative 13 in 83% yield. Opening of the 1,6-anhydro ring with tri-
fluoroacetic acid in acetic anhydride then gave the stereoselectively
mono-deuteriated glucosamine derivative 14 in 83% yield.
Subsequent treatment of 14 with HCl in hot methanol gave the
methyl
a- and b-glycosides 15a and 15b in 42 and 25% yield,
respectively, which, on hydrogenolysis over Pearlman's catalyst
gave the target compounds 6S-2H-4 and 6S-2H-5 in good yield
(Scheme 2).
Sulfonylation of anomers 15a and 15b gave the 6-O-sulfonate
esters 16 and 18 in excellent yield, which, on exposure to sodium
azide in hot aqueous DMF gave the respective 6-azido-6-deoxy
derivatives 17 and 19 in excellent yield with clean inversion of
configuration (Scheme 3). Finally, hydrogenolysis of 17 and 19
afforded the methyl 2,6-dideoxy-2,6-diamino-a- and b-glucosides
6R-2H-2 and 6R-2H-3 in good yield (Scheme 3). Authentic samples
of non-deuteriated 2, 4 and 5 were either commercial or were
obtained by literature methods, while an authentic sample of 3 was
prepared by routine methods as described in Scheme 4 from the
Fig. 1. Staggered Conformations of the Side Chain in the Hexopyranoses Illustrated for
Methyl -D-glucopyranoside.
known [12] N-benzyloxycarbonyl derivative 20 of methyl
b-D-
a
glucosaminide.
1H-NMR spectra of 2e5, of 6R-2H-2 and 6R-2H-3, and of 6S-2H-4
and 6S-2H-5 were obtained in D2O at pH 5 and pH 11 leading to the
complete assignment of the H5, H6R and H6S resonances and their
2. Results and discussion
associated coupling constants laid out in Tables
respectively.
The H5-H6R coupling constants for 2 and 3 of 8.8 and 9.2 Hz and
the corresponding and H5-H6S coupling constants of 2.8 and 2.9 Hz
in aqueous acetic acid (Table 1), are comparable to those (~9.2 and
1 and 2,
Although shorter methods for the stereospecific introduction of
deuterium into the hydroxymethyl group of the hexopyranosides
based on asymmetric reduction of the corresponding aldehydes
exist [4e6], we have preferred to adapt the earlier Meguro route
based on the photobromination and deuteriodebromination of 1,6-
anhydrohexose derivatives because of its unambiguous nature
[7e10]. Accordingly, white light stimulated bromination of 1,6-
anhydro-D-glucopyranose triacetate 6 in a,a,a-trifluorotoluene
[11] gave 78% of the 6-exo-bromide 7, which was reduced with
tributyltin deuteride in hot toluene to give the 6-exo-deuterio 1,6-
3.1 Hz) recorded for methyl 6-amino-6-deoxy-b-D-glucopyranoside
1 by Bock and Duus at pH 4.75 [1]. Accordingly, we conclude that i)
the H6R and H6S resonances of 1 were correctly assigned and ii)
that the side chains of 1, 2 and 3 populate very similar equilibrium
mixtures of conformers, calculated on the basis of the Haasnoot-
Altona equation [13] to be a 16:80:4 gg:gt:tg ratio for 1 by Bock
Scheme 1. Synthesis of the 6S-deuterio-2-azido-2-deoxyglucose derivative 14.

12
T. Kato et al. / Carbohydrate Research 448 (2017) 10e17
Scheme 2. Synthesis of the 6S-deuteriated methyl a- and b-glucosaminides 4 and 5.
Scheme 3. Synthesis of the 6R-deuteriated methyl a- and b-glucosaminides 2 and 3.
Scheme 4. Preparation of Unlabeled 3.
Table 1
constants for 2 and 3, indicating that the predominance of the gt
conformation for these glycosides is maintained in basic solution.
Similarly, the H5-H6R and H5-H6S coupling constants for 4 and 5 in
both aqueous acetic acid (Table 1) and basic D2O (Table 2) deviate
Diagnostic chemical shifts and coupling constants for compounds 2e5 and their
selectively monodeuteriated analogs in D2O at pH 5.
2/6R-2H-2
3/6R-2H-3
4/6S-2H-4
5/6S-2H-5
little from those reported [1] for the methyl a-D- and b-D-gluco-
pyranosides in water indicating that similar side chain conforma-
tional preferences are adopted and approximate the 53:47:0 and
d
d
d
H5
H6R
H6S
3.84/3.84
3.13/-
3.42/3.41
8.8/-
2.8/2.9
13.2/-
3.65/3.65
3.12/-
3.44/3.42
9.2/-
2.9/2.8
13.3/-
3.53/3.53
3.62/3.60
3.71/-
4.9/5.1
2.2/-
3.34/3.34
3.59/3.57
3.77/-
5.5/5.5
1.8/-
J5,6R (Hz)
J5,6S (Hz)
J6S,6R (Hz)
47:53:0 gg:gt:tg ratios computed by Bock and Duus for the
glucosides [1]. We further conclude that replacement of the 2-
hydroxy group in the - and -glucopyranosides and their 6-
a- and b-
13.2/-
12.1/-
a
b-D
amino derivatives by a protonated amino group has only a mini-
mal effect on the side chain conformation. The increased popula-
tion of the gt conformer in the 6-amino series in acidic media
compared to the corresponding 6-hydroxy compounds is consis-
tent with the predominantly gauche conformation (~92%) adopted
by ethanol amine hydrochloride in aqueous solution determined by
Roberts and coworkers [14]. The major gt conformation of 2 and 3
in aqueous base is also consistent with the predominant gauche
conformation of neutral ethanol amine as determined by NMR
spectroscopy [14], and computationally [15], and as determined by
vibrational spectroscopy in the gas phase [16].
Table 2
Diagnostic chemical shifts and coupling constants for compounds 2e5 and their
selectively monodeuteriated analogs in D2O at pH 11.
2/6R-2H-2
3/6R-2H-3
4/6S-2H-4
5/6S-2H-5
d
d
d
H5
H6R
H6S
3.55/3.55
2.58/-
3.00/3.01
9.1/-
2.6/2.7
12.5/-
3.38/3.38
2.60/-
3.03/3.01
9.0/-
2.7/2.7
12.7/-
3.32/3.33
3.45/3.44
3.57/-
5.3/5.3
2.2/-
3.18/3.17
3.39/3.39
3.60/-
5.8/5.7
2.1/-
J5,6R (Hz)
J5,6S (Hz)
J6S,6R (Hz)
12.5/-
12.4/-
Regarding the influence of anomeric configuration on side chain
conformation, comparison of the coupling constant data for 2 and 3
in both acidic and basic media (Tables 1 and 2) reveals only minor
differences suggesting that the side chain amine is the determining
and Duus [1]. On going from pH 5 (Table 1) to pH 11 (Table 2) there
is only minimal change in the H5-H6R and H5-H6S coupling

T. Kato et al. / Carbohydrate Research 448 (2017) 10e17
13
factor. For 3 and 4, which lack the 6-amino group, the changes,
while still modest, are more pronounced in both acidic and basic
media (Tables 1 and 2) and are consistent with a slightly higher
population of the gt conformer for the equatorial anomer. This
pattern follows that observed for the anomers of glucopyranosides
by multiple groups [1,17,18].
cooled to ambient temperature then was concentrated under
reduced pressure. The residue was dissolved in a minimum of
toluene and dichloromethane (1:1 mixture) and was purified by
column chromatography (hexane:ethyl acetate 3:1 to 1:1) to give
the white solid 6-(S)-deuterio
8 (5.5 g, 0.019 mol, 82%).
½
a 2D3
ꢀ
¼ ꢁ39.1 (c ¼ 0.15), 1H NMR (600 MHz, CDCl3):
d
¼ 5.36 (br s,
1H, H1); 4.75 (m, 1H, H3); 4.55 (br s, 1H, H4); 4.52 (br s, 1H, H5);
4.49 (br s, 1H, H2): 3.99 (s, 1H, H6); 2.07 (s, 3H, CH3); 2.05 (s, 3H,
3. Conclusion
CH3); 2.02 (s, 3H, CH3). 13C NMR (150 MHz, CDCl3):
d
¼ 169.87,
The side chains of both anomers of the methyl glycosides of 2,6-
diamino-2,6-dideoxy-D-glucopyranose very predominantly adopt
the gt conformation in aqueous solution at pH 5 and pH 11 as
revealed by the stereospecific synthesis of 6-monodeuterio iso-
topomers. In contrast, anomers of the methyl glycosides of 2-
169.51, 168.95 (-COCH3 x3); 99.04 (C1); 73.54 (C5); 70.27 (C4);
69.59 (C3); 69.03 (C2); 64.95 (t, J C-D ¼ 23.6 Hz, C6); 20.86 (-COCH3);
20.76 (-COCH3 x2). ESI-HRMS: m/z calcd for C12H15DO8Na
[M þ Na]þ 312.0806, found 312.0815.
amino-2-deoxy-D-glucopyranoside populate, like the methyl gly-
4.1.3. 1,6-Anhydro-2,4-di-O-p-toluenesulfonyl-6-(S)-deuterio-b-D-
cosides of glucopyranose itself, approximately 1:1 mixtures of the
gg and gt conformations. The preference for the gt conformation in
the 2,6-diamino series is therefore a consequence of the 6-amino
group, and is consistent with the gauche conformation adopted
by 2-aminoethanol under similar conditions.
glucopyranose (9)
Compound 8 (6.0 g 0.021 mol) was dissolved in dry methanol
(60 mL) and treated with sodium methoxide (200 mg, 0.37 mmol)
at ambient temperature, with monitoring by TLC (chlor-
oform:methanol 5:1, Rf ¼ 0.4). After 5 h the reaction mixture was
neutralized with Amberlite® IR120, then filtered, concentrated, and
dried under reduced pressure. The residue was dissolved in pyri-
dine (18 mL) and treated with a solution of p-toluenesulfonyl
chloride (8.7 g, 0.046 mol) in 1:1 pyridine:chloroform (55 mL) in an
ice bath. The reaction mixture was allowed to warm to ambient
temperature, and stirred for 12 h, with monitoring by TLC (hex-
ane:ethyl acetate 1:1, Rf ¼ 0.6). The reaction mixture was quenched
with methanol (5 mL), then concentrated under reduced pressure.
The residue was filtered through silica gel (hexane:ethyl acetate
2:1) and concentrated under vacuum to give 9 as a light yellow oil
(8.1 g, 0.017 mol, 82%), whose spectra were consistent with the
literature [22e24] for the non-deuteriated isotopomer and used in
the next reaction without further purification. ESI-HRMS: m/z calcd
for C20H21DO9S2Na [M þ Na]þ 494.0666, found 494.0659.
4. Experimental section
4.1. General
All reactions were conducted under an atmosphere of nitrogen
or argon. Extracts were dried over sodium sulfate and chromatog-
raphy was carried out over silica gel. Sephadex chromatography
was carried out over Sephadex C25. Specific rotations were
measured on an automatic polarimeter with a path length of 10 cm
in chloroform solution unless otherwise stated. High-resolution
(HRMS) mass spectra were recorded in the electrospray mode us-
ing a time of flight mass analyzer (ESI-TOF).
4.1.1. 1,6-Anhydro-2,3,4-tri-O-acetyl-6-(S)-bromo-b-D-
glucopyranose (7)
1,6-Anhydro-2,3,4-tri-O-acetyl-
(10.0 g, 0.035 mol) was dissolved in
4.1.4. 1,6; 3,4-bisanhydro-2-O-p-toluenesulfonyl-6-(S)-deuterio-
-galactopyranose (10)
A solution of 9 (7.0 g, 0.015 mol) in dry methanol (140 mL) and
b-
b
-
D
-glucopyranose
6
[19]
D
a,a,a
-trifluoromethylbenzene
(600 mL) and N-bromosuccinimide (24.9 g, 0.14 mol) was added at
ambient temperature. The reaction mixture was heated to reflux
with irradiation by a 300 W incandescent bulb for 6 h, monitoring
by TLC (Hexane:ethyl acetate 1:1, Rf ¼ 0.55). After completion, the
reaction mixture was concentrated under reduced pressure, diluted
with ethyl acetate (200 mL) and was washed with saturated
aqueous sodium bicarbonate. The aqueous layer was extracted with
ethyl acetate twice and the combined organic layer was washed
with brine, dried, and concentrated to dryness. The residual syrup
was purified by silica gel column chromatography (hexane:ethyl
acetate 5:1 to 2:1) to give the 6-(S)-bromo compound 7 [20] as a
was treated with 0.6 M methanolic sodium methoxide (94 mL) in
an ice bath. The reaction mixture was allowed to warm to ambient
temperature and stirred for 10 h, with monitoring by TLC (hex-
ane:ethyl acetate 1:1, Rf ¼ 0.65). After completion the reaction
mixture was concentrated and the residue taken up in ethyl acetate
(150 mL) and washed with water (100 mL). The aqueous layer was
extracted with ethyl acetate and the combined organic layer
washed with brine, dried, and concentrated. The residue was pu-
rified by column chromatography (hexane:ethyl acetate 3:1 to 1:1)
to give 10 as a white amorphous solid (3.6 g, 0.012 mol, 77%), whose
spectra were consistent with the literature for the non-deuteriated
colorless syrup (9.9 g, 0.027 mol, 78%). ½a D23
ꢀ
¼ ꢁ101.1 (c ¼ 1.0), 1H
isotopomer [23,24]. ½a D23
ꢀ
¼ ꢁ43.1 (c ¼ 1.2). 1H NMR (400 MHz,
NMR (600 MHz, CDCl3):
d
¼ 6.41 (s, 1H, H6); 5.77 (br s,1H, H1); 4.83
CDCl3):
d
¼ 7.84 (d, J ¼ 8.3 Hz, 2H, arom.); 7.38 (d, J ¼ 8.3 Hz, 2H,
(br s, 1H, H3); 4.79 (br s, 1H, H5); 4.66 (br s, 1H, H4); 4.50 (br s, 1H,
arom.); 5.16 (br s, 1H, H1); 4.83 (d, J5-4 ¼ 4.8 Hz, 1H, H5); 4.39 (br s,
1H, H2); 3.92 (s, 1H, H6); 3.60 (t, J4-3 ¼ 4.4 Hz, J4-5 ¼ 4.4 Hz, 1H, H4);
H2); 2.11 (s, 3H, CH3); 2.09 (s, 3H, CH3); 2.05 (s, 3H, CH3). 13C NMR
(150 MHz, CDCl3):
d
¼ 169.76, 169.50, 168.87 (-COCH3 x3); 102.08
3.13 (dd, J3-4 ¼ 4.0 Hz, J3-N.D. ¼ 1.5 Hz, 1H, H3); 2.46 (s, 3H, CH3). 13
C
(C1); 84.19 (C5); 79.36 (C6); 69.29 (C3); 67.93 (C2); 67.67 (C4);
20.77 (CH3 x2); 20.71 (CH3). ESI-HRMS: m/z calcd for C12H15O8BrNa
[M þ Na]þ 388.9848, found 388.9853.
NMR (100 MHz, CDCl3):
d
¼ 146.63, 132.80, 138.19, 127.98 (arom.);
98.05 (C1); 71.78 (C2); 71.53 (C5); 64.44 (t, JC-D ¼ 24.4 Hz, C6); 52.82
(C4); 47.63 (C3); 21.71 (CH3). ESI-HRMS: m/z calcd for
C
13H13DO6SNa [M þ Na]þ 322.0472, found 322.0481.
4.1.2. 1,6-Anhydro-2,3,4-tri-O-acetyl-6-(S)-deuterio-b-D-
glucopyranose (8) [7]
4.1.5. 1,6-Anhydro-4-O-benzyl-2-O-p-toluenesulfonyl-6-(S)-
deuterio- -glucopyranose (11)
A solution of epoxide 10 (4.0 g, 0.013 mol) in benzene (80 mL),
was treated with benzyl alcohol (7 mL, 0.04 mol) and 35% boron
trifluoride diethyl etherate (1 mL, 0.002 mol) at ambient temper-
ature. The solution was heated to 50 ꢂC with stirring for 12 h and
Compound 7 (8.3 g, 0.023 mol), tributyltin deuteride prepared
according to Neumann [21] (21.0 g, 0.069 mol) and azobisisobu-
tyronitrile (365 mg, 0.0023 mol) were dissolved in toluene
(830 mL) and heated to reflux, with monitoring by TLC (hex-
ane:ethyl acetate 1:1, Rf ¼ 0.65). After 1 h, the reaction mixture was
b-D

14
T. Kato et al. / Carbohydrate Research 448 (2017) 10e17
monitored by TLC (hexane:ethyl acetate 1:1, Rf ¼ 0.55). After
completion the reaction mixture was poured into saturated
aqueous sodium bicarbonate (100 mL), the aqueous layer was
separated and extracted with ethyl acetate. The combined organic
layer was washed with brine, dried, and concentrated under
reduced pressure. The residue was purified by column chroma-
tography (hexane:ethyl acetate 3:1 to 1:1) to give 11 (4.1 g,
0.010 mol, 78%) as a white solid, whose spectra were consistent
with the literature for the non-deuteriated isotopomer [23,24].
2:1) to give the colorless oil 13 (2.2 g, 0.0060 mol, 83%), whose
spectra were consistent with the literature for the non-deuteriated
isotopomer [25]. ½a D23
ꢀ
¼ þ41.2 (c ¼ 0.5). 1H NMR (600 MHz, CDCl3):
d
¼ 7.40e7.26 (m,10H, arom.); 5.49 (s,1H, H1); 4.62 (d, J5-4 ¼ 2.2 Hz,
1H, H5); 4.61e4.48 (m, 4H, CH2 x2); 3.99 (s, 1H, H6); 3.65 (br s, 1H,
H3); 3.38 (br s, 1H, H4); 3.28 (br s, 1H, H2). 13C NMR (150 MHz,
CDCl3):
d
¼ 137.39, 137.24, 128.57, 128.56, 128.07, 128.02, 127.91,
127.79 (arom.); 100.53 (C1); 76.18 (C3); 75.84 (C4); 74.31 (C5);
72.35 (CH2); 71.33 (CH2); 65.01 (t, JC-D ¼ 23.6 Hz, C6); 59.88 (C2).
ESI-HRMS: m/z calcd for C20H20DN3O4Na [M þ Na]þ 391.1493,
found 391.1488.
½
a 2D3
ꢀ
¼ ꢁ22.0 (c ¼ 2.0). 1H NMR (600 MHz, CDCl3):
d
¼ 7.81 (d,
J ¼ 8.4 Hz, 2H, arom.); 7.37e7.25 (m, 7H, arom.); 5.31 (br s, 1H, H1);
4.67 (d, JCH2a-CH2b ¼ 12.1 Hz, 1H, CH2a); 4.59 (d, JCH2b-CH2a ¼ 12.1 Hz,
1H, CH2b); 4.55 (s, 1H, H5); 4.21 (d, J2-3 ¼ 3.4 Hz, 1H, H2); 3.96 (t, J3-
4.1.8. 1,6-Di-O-acetyl-2-azido-2-deoxy-3,4-di-O-benzyl-6-(S)-
deuterio-a-D-glucopyranose (14)
¼ 3.6 Hz, J3-4 ¼ 3.6 Hz, 1H, H3); 3.87 (s, 1H, H6); 3.31 (d, J4-
2
¼ 3.6 Hz, 1H, H4); 2.43 (s, 3H, CH3). 13C NMR (150 MHz, CDCl3):
Compound 13 (2.1 g, 0.0057 mol) was dissolved in acetic an-
hydride (33 mL), treated with trifluoroacetic acid (3.3 mL) at
ambient temperature, and stirred for 1.5 h, with monitoring by TLC
(hexane:ethyl acetate 2:1, Rf ¼ 0.40). The reaction mixture was
poured into saturated aqueous sodium carbonate (300 mL), and
extracted with ethyl acetate. The combined organic layer was
washed with brine, dried, and concentrated. The residue was pu-
rified by column chromatography (hexane:ethyl acetate 4:1 to 2:1)
to give the colorless oil 14 (2.2 g, 4.73 mmol, 83%), whose spectra
were consistent with the literature for the non-deuteriated iso-
3
d
¼ 145.29, 137.49, 133.06, 129.98, 128.52, 127.96, 127.84 (arom.);
99.88 (C1); 79.18 (C2); 78.40 (C4); 75.13 (C5); 71.73 (CH2); 70.12
(C3); 66.10 (t, JC-D ¼ 23.5 Hz, C6); 21.69 (CH3). ESI-HRMS: m/z calcd
for C20H21DO7SNa [M þ Na]þ 430.1047, found 430.1039.
4.1.6. 1,6; 2,3-Bisanhydro-4-O-benzyl-6-(S)-deuterio-b-D-
mannopyranose (12)
A solution of 11 (4.3 g, 0.011 mol) in chloroform (25 mL) was
treated with 1.0 M methanolic sodium methoxide (25 mL) in an ice
bath, then was allowed to warm to ambient temperature, and
stirred for 5 h with monitoring by TLC (hexane:ethyl acetate 1:1,
Rf ¼ 0.60). The reaction mixture was poured into a mixture of water
(150 mL) and chloroform (150 mL). The aqueous layer was sepa-
rated and extracted with chloroform and the combined organic
layer was washed with brine, dried and concentrated. The residue
was purified by column chromatography (hexane:ethyl acetate 3:1
to 1:1) to give 12 (2.3 g, 8.14 mmol, 88%) as a colorless oil, whose
spectra were consistent with the literature for the non-deuteriated
topomer [26]. ½a D23
ꢀ
¼ þ58.1 (c ¼ 1.2). 1H NMR (600 MHz, CDCl3):
d
¼ 7.45e7.26 (m, 10H, arom.); 6.23 (d, J1-2 ¼ 3.7 Hz, 1H, H1); 4.94
(d, JCHa-CHb ¼ 10.3 Hz, 1H, -O-CH2aPh); 4.91; (d, JCHb-CHa ¼ 10.3 Hz,
1H, -O-CH2bPh); 4.88 (d, JCHa’-CHb’ ¼ 10.6 Hz, 1H, -O-CH2a’Ph); 4.59
(d, JCHb’-CHa’ ¼ 10.6 Hz, 1H, -O-CH2b’Ph); 4.25 (d, J6-5 ¼ 4.0 Hz, 1H,
H6); 3.98 (t, J3-2 ¼ J3-4 ¼ 9.2 Hz, 1H, H3); 3.93 (dd, J5-4 ¼ 9.4 Hz, J5-
¼ 4.1 Hz, 1H, H5); 3.64 (t, J4-3 ¼ J4-5 ¼ 9.2 Hz, 1H, H4); 3.60 (dd, J2-
6
¼ 3.7 Hz, J2-3 ¼ 9.6 Hz, 1H, H2); 2.15 (s, 3H, CH3); 2.03 (s, 3H, CH3).
1
13C NMR (150 MHz, CDCl3):
d
¼ 170.53, 168.75 (2C, C¼O x2); 137.44,
isotopomer [23]. ½a D23
ꢀ
¼ ꢁ28.3 (c ¼ 1.3). 1H NMR (600 MHz, CDCl3):
137.22 (2C, arom.); 128.62, 128.57, 128.22, 128.11, 128.08 (10C,
arom.); 90.37 (C1); 80.55 (C3); 76.87 (C4); 75.68, 75.29 (2C, -CH2-
x2); 71.22 (C5); 62.71 (C2); 62.04 (t, JC-D ¼ 23.0 Hz, C6); 20.94, 20.78
(2C, -COCH3). ESI-HRMS: m/z calcd for C24H26DN3O7Na [M þ Na]þ
493.1809, found 493.1798.
d
¼ 7.40e7.20 (m, 5H, arom.); 5.69 (d, J1-2 ¼ 3.1 Hz, 1H, H1); 4.72 (s,
2H, CH2); 4.49 (br s,1H, H5); 3.64 (br s,1H, H6); 3.43 (t, J2-1 ¼3.4 Hz,
J2-3 ¼ 3.3 Hz, 1H, H2); 3.18 (dd, J3-2 ¼ 3.6 Hz, J3-N.D. ¼ 1.5 Hz, 1H, H3).
13C NMR (150 MHz, CDCl3):
d
¼ 137.27, 128.61, 128.12, 127.82
(arom.); 97.57 (C1); 73.31 (C4); 72.13 (CH2); 71.53 (C5); 65.46 (t, JC-
¼ 23.6 Hz, C6); 54.37 (C2); 47.80 (CH3). ESI-HRMS: m/z calcd for
4.1.9. Methyl 2-azido-2-deoxy-3,4-di-O-benzyl-6-(S)-deuterio-
a-
D
C
13H13DO4Na [M þ Na]þ 258.0853, found 258.0862.
and -glucopyranose (15 ) and (15
b
-D
a
b)
The 1,6-di-O-acatate 14 (520 mg, 0.318 mmol) was dissolved in
10% w/w methanolic hydrochloric acid (20 mL, prepared by addi-
tion of acetyl chloride (3.0 mL) to dry methanol (17 mL) in an ice
bath) at ambient temperature and stirred for 8 h at 75 ꢂC, with
monitoring by TLC (hexane:ethyl acetate 2:1, Rf ¼ 0.35 for 15ab and
Rf ¼ 0.15 for the intermediate 1,6-diol) and ESI mass spectrometry.
After conversion of the intermediate pyranose to the methyl gly-
cosides, the reaction mixture was concentrated under reduced
pressure, taken up in ethyl acetate and washed with saturated
aqueous sodium bicarbonate. The aqueous layer was extracted with
ethyl acetate, and the combined organic layer was washed with
brine, dried, and concentrated. The residue was dissolved in the
minimum volume of toluene for purification by column chroma-
tography (hexane:ethyl acetate 4:1 to 2:1) which gave the colorless
4.1.7. 1,6-Anhydro-2-azido-2-deoxy-3,4-di-O-benzyl-6-(S)-
deuterio- -glucopyranose (13)
b-D
A solution of 12 (2.0 g, 0.0085 mol) 9:1 water:DMF (40 mL), was
treated with sodium azide (5.5 g, 0.085 mol) and ammonium
chloride (3.4 g, 0.064 mol), and heated to reflux with stirring for
10 h, and monitoring by TLC (hexane:ethyl acetate 1:1, Rf ¼ 0.50).
The reaction mixture was concentrated under reduced pressure
and the residue taken up in ethyl acetate (100 mL) and washed with
water. The aqueous layer was extracted with ethyl acetate, and the
combined organic layer was washed with brine, dried, and
concentrated. The concentrate (2.0 g, 0.0072 mol, ESI-HRMS: m/z
calcd for C13H14DN3O4Na [M þ Na]þ 301.1023, found 301.1015) was
dissolved in tetrahydrofuran (20 mL) and treated with sodium
hydride (60% dispersion in oil, 430 mg, 0.011 mol) at ambient
temperature. After stirring for 0.5 h benzyl bromide (1.8 mL,
0.014 mol) was added at ambient temperature and the reaction
mixture stirred for 3 h with monitoring by TLC (hexane:ethyl ace-
tate 2:1, Rf ¼ 0.45). After completion methanol (1 mL) was added
and the volatiles were removed under reduced pressure. The res-
idue was dissolved in ethyl acetate and washed with water. The
aqueous layer was extracted with ethyl acetate, and the combined
organic layer washed, dried, and concentrated. The residue was
purified by column chromatography (hexane:ethyl acetate 5:1 to
oils 15a (53.4 mg, 0.134 mmol, 42%) and 15b (32.0 mg, 0.080 mmol,
25%), whose spectra were consistent with the literature for the
non-deuteriated isotopomers [27,28]. 15
a
: ½a 2D3
¼ þ78.3 (c ¼ 1.0).
ꢀ
1H NMR (600 MHz, CDCl3):
d
¼ 7.43e7.27 (m, 10H, arom.);
4.93e4.85 (m, 3H, -CH2a,bPh, -CH2a’Ph); 4.78 (d, J1-2 ¼ 3.7 Hz, 1H,
H1); 4.68 (d, JCH2b’-CH2a’ ¼ 11.0 Hz, 1H, -CH2b’Ph); 4.01 (dd, J3-
¼ 9.9 Hz, J3-4 ¼ 9.0 Hz, 1H, H3); 3.72 (d, J6-5 ¼ 3.9 Hz, 1H, H6); 3.70
2
(dd, J5-4 ¼ 9.5 Hz, J5-6 ¼ 4.0 Hz, 1H, H5); 3.62 (t, J4-3 ¼ J4-5 ¼ 9.3 Hz,
1H, H4); 3.41 (s, 3H, -OCH3); 3.60 (dd, J2-1 ¼ 3.7 Hz, J2-3 ¼ 10.1 Hz,
1H, H2). 13C NMR (150 MHz, CDCl3)
d
¼ 137.83, 137.79 (2C, arom.);

T. Kato et al. / Carbohydrate Research 448 (2017) 10e17
15
128.72e127.72 (10C, arom.); 98.69 (C1); 80.35 (C3); 77.86 (C4);
75.52, 75.04 (2C, -CH2- x2); 71.19 (C5); 63.67 (C2); 61.16 (t, JC-
4.1.12. Methyl 2-azido-2-deoxy-3,4-di-O-benzy-6-O-p-
toluenesulfonyl-6-(S)-deuterio- -glucopyranoside (16)
A solution of 15 (55.0 mg, 0.137 mmol) in pyridine (0.5 mL) and
a-D
¼ 22.4 Hz, C6); 55.24 (-OCH3). ESI-HRMS: m/z calcd for
a
D
C
½
21H24DN3O5Na [M
ꢀ
þ
Na]þ 423.1755, found 423.1761.15
b:
p-toluenesulfonyl chloride (39.2 mg, 0.206 mmol) was stirred at
ambient temperature for 2 h, with monitoring by TLC (hexane:ethyl
acetate 2:1, Rf ¼ 0.40). The reaction mixture was diluted with ethyl
acetate and washed with 1 N hydrochloric acid. The aqueous layer
was extracted with ethyl acetate and the combined extracts were
washed with saturated aqueous sodium bicarbonate and brine,
dried, concentrated and purified by silica gel column chromatog-
raphy (hexane:ethyl acetate 4:1 to 2:1) to give the tosylate 16 as a
a 2D3
¼ ꢁ26.9 (c ¼ 0.3). 1H NMR (600 MHz, CDCl3):
¼ 7.39e7.27 (m,
d
10H, arom.); 4.89 (d, JCHa-CHb ¼ 10.6 Hz, 1H, -CH2aPh); 4.85; (d, JCHa’-
¼ 11.0 Hz, 1H, -CH2a’Ph); 4.83 (d, JCHb-CHa ¼ 10.6 Hz, 1H,
CHb’
-CHbPh); 4.66 (d, JCHb’-CHa’ ¼ 11.0 Hz, 1H, -CH2b’Ph); 4.21 (d, J1-
¼ 8.1 Hz, 1H, H1); 3.71 (d, J6-5 ¼ 4.4 Hz, 1H, H6); 3.59 (t, J4-3 ¼ J4-
2
¼ 9.4 Hz, 1H, H4); 3.57 (s, 3H, -OCH3); 3.46 (t, J3-2 ¼ J3-4 ¼ 9.5 Hz,
5
1H, H3); 3.37 (dd, J2-1 ¼ 8.1 Hz, J2-3 ¼ 9.7 Hz, 1H, H2); 3.33 (dd, J5-
¼ 9.6 Hz, J5-6 ¼ 4.4 Hz, 1H, H5). 13C NMR (150 MHz, CDCl3):
colorless oil (58.4 mg, 0.105 mmol, 77%). ½a D23
ꢀ
¼ þ68.1 (c ¼ 0.5). 1H
4
d
¼ 137.78, 137.69 (2C, arom.); 128.60e127.87 (10C, arom.); 103.01
NMR (600 MHz, CDCl3):
d
¼ 7.78 (d, J ¼ 8.4 Hz, 2H, arom); 7.38e7.27
(C1); 82.93 (C3); 77.35 (C4); 75.57(-CH2PH); 75.17 (C5); 75.12
(-CH2PH); 66.32 (C2); 61.33 (t, JC-D ¼ 23.6 Hz, C6); 57.31 (-OCH3).
ESI-HRMS: m/z calcd for C21H24DN3O5Na [M þ Na]þ 423.1755,
found 423.1747.
(m, 10 H, arom); 7.18 (dd, J ¼ 2.2 Hz, 7.7 Hz, 2H, arom); 4.88 (d, JCH2a-
¼ 10.6 Hz, 1H, -CH2aPh); 4.82 (d, JCH2b-CH2a ¼ 10.6 Hz, 1H,
CH2b
-CH2bPh); 4.81 (d, JCH2a’-CH2b’ ¼ 10.6 Hz, 1H, -CH2a’Ph); 4.47 (d, J1-
¼ 3.7 Hz, 1H, H1); 4.47 (d, JCH2b’-CH2a’ ¼ 10.6 Hz, 1H, -CH2b’Ph); 4.21
2
(d, J6-5 ¼ 4.4 Hz, 1H, H6); 3.94 (dd, J3-2 ¼ 9.9 Hz, J3-4 ¼ 9.0 Hz, 1H,
H3); 3.80 (dd, J5-4 ¼ 10.1 Hz, J5-6 ¼ 4.4 Hz, 1H, H5); 3.51 (dd, J4-
4.1.10. Methyl 6-(S)-deuterio-a-D-glucosaminide monoacetate
¼ 9.1 Hz, J4-5 ¼ 10.1, 1H, H4); 3.36 (s, 3H, -OCH3); 3.34 (dd, J2-
3
(6S-2H-4)
¼ 3.7 Hz, J2-3 ¼ 10.1 Hz, 1H, H2); 2.41 (s, 3H, -SO2PhCH3). 13C NMR
1
To a solution of 15a (30 mg, 0.075 mmol) in 1,4-dioxane (1 mL)
(150 MHz, CDCl3)
d
¼ 144.97, 137.60, 137.42, 132.81 (4C, arom);
was added the same weight of 10% palladium hydroxide on acti-
vated charcoal. The reaction mixture was stirred 6 H under 1 atm of
hydrogen atmosphere. Water (0.1 mL) was then added and the
pressure adjusted to 3 atm of hydrogen. After stirring for 12 h the
catalyst was filtered off and washed with water until the washings
gave a negative ninhydrin test. The filtrate was concentrated and
dried under reduced pressure. The residue was dissolved in
methanol (0.5 mL) and slurried with silica gel. The methanol was
removed in vacuo and the powder applied to a silica gel column
that was eluted with chloroform:methanol:ammonium hydroxide
3:1:0.5, Rf ¼ 0.25. The concentrated fractions were taken up in the
minimum volume of 10% aqueous acetic acid and purified by
Sephadex C-25 column (eluent: water then 2% ammonium hy-
130.00e127.75 (14C, ar0om); 98.53 (C1); 80.30 (C3); 77.43 (C4);
75.48 (CH2); 75.02 (CH2 ); 68.90 (C5); 67.97 (t, JC-D ¼ 22.4 Hz, C6);
63.40 (C2); 55.40 (-OCH3); 21.64 (PhCH3). ESI-HRMS: m/z calcd for
C
28H30DN3O7SNa [M þ Na]þ 577.1843, found 577.1851.
4.1.13. Methyl 2,6-diazido-2,6-dideoxy-3,4-di-O-benzyl-6-(R)-
deuterio- -glucopyranoside (17)
a-D
A solution of 16 (85.0 mg, 0.153 mmol) in 9:1 N,N-dime-
thylformamide:water (1.2 mL) mixture was treated sodium azide
(119 mg, 1.84 mol) and heated to 85 ꢂC for 8 h with stirring, and
monitoring by TLC (hexane:ethyl acetate 3:1, Rf ¼ 0.45). On
completion the reaction mixture was diluted with ethyl acetate and
washed with water. The aqueous layer was extracted with ethyl
acetate and the combined organic layer was washed with brine,
dried, concentrated, and purified by column chromatography
(hexane:ethyl acetate 6:1 to 3:1) to give 17 as white foam (52.2 mg,
droxide). Glacial acetic (50
mL) acid was added to the product-
containing fractions, followed by drying under vacuum to give
the title compound 6S-2H-4 as colorless oil (10.4 mg, 0.041 mmol,
55%), whose non-deuteriated isotopomer was previously reported
0.123 mmol, 80%). ½a D23
ꢀ
¼ þ59.3 (c ¼ 0.5). 1H NMR (600 MHz,
as the free base [29]. ½a D23
ꢀ
¼ þ152.3 (c ¼ 0.1, water). 1H NMR
CDCl3):
d
¼ 7.42e7.22 (m, 10H, arom); 4.89 (d, JCH2a-CH2b ¼ 10.6 Hz,
(600 MHz, D2O):
d
¼ 4.85 (d, J1-2 ¼ 3.7 Hz, 1H, H1); 3.86 (t, J3-2 ¼ J3-
1H, -CH2aPh); 4.88 (d, JCH2a’-CH2b’ ¼ 11.0 Hz, 1H, -CH2a’Ph); 4.84 (d,
JCH2b-CH2a ¼ 10.6 Hz, 1H, -CH2bPh); 4.80 (d, J1-2 ¼ 3.4 Hz, 1H, H1);
4.59 (d, JCH2b’-CH2a’ ¼ 11.0 Hz, 1H, -CH2b’Ph); 3.97 (dd, J3-2 ¼ 10.3 Hz,
J3-4 ¼ 9.2 Hz, 1H, H3); 3.81 (dd, J5-4 ¼ 9.8 Hz, J5-6 ¼ 2.2 Hz, 1H, H5);
3.52 (t, J4-3 ¼ J4-5 ¼ 9.5, 1H, H4); 3.46 (d, J6-5 ¼ 2.2 Hz, 1H, H6); 3.44
¼ 9.7 Hz, 1H, H3); 3.60 (d, J6-5 ¼ 5.1 Hz, 1H, H6); 3.53 (dd, J5-
¼ 9.8 Hz, J5-6 ¼ 5.1 Hz, 1H, H5); 3.31 (t, J4-3 ¼ J4-5 ¼ 9.7 Hz, 1H,
4
4
H4); 3.28 (s, 3H, -OCH3); 3.16 (dd, J2-1 ¼ 3.7 Hz, J2-3 ¼ 9.9 Hz, 1H,
H2). 13C NMR (150 MHz, D2O):
d
¼ 176.63 (-C(O)OH); 96.03 (C1);
71.72 (C6); 69.82 (C3); 69.35 (C4); 59.84 (t, JC-D ¼ 22.0 Hz, C6); 55.13
(-OCH3); 53.87 (C2); 20.35 (CH3C(O)OH). ESI-HRMS: m/z calcd for
C7H15DNO5 [MþH]þ 195.1091, found 195.1098.
(s, 3H, -OCH3); 3.42 (dd, J2-1 ¼ 3.5 Hz, J2-3 ¼ 10.3 Hz, 1H, H2). 13
C
NMR (150 MHz, CDCl3):
d
¼
135.05, 135.01 (2C, arom);
126.08e125.14 (10C, arom); 96.00 (C1); 77.74 (C3); 76.21 (C4);
0
72.97(CH2); 72.60 (CH2 ); 67.82 (C5); 61.03 (C2); 52.84 (-OCH3);
4.1.11. Methyl 6-(S)-deuterio-
b
-
D
-glucosaminide monoacetate
48.19 (t, JC-D ¼ 21.9 Hz, C6). ESI-HRMS: m/z calcd for C21H23DN6O4Na
(6S-2H-5)
[M þ Na]þ 448.1819, found 448.1811.
Compound 15
b (26.1 mg, 0.065 mmol) was subjected to
hydrogenolysis and purification analogously to 15
a
to give the title
4.1.14. Methyl 2,6-diamino-2,6-dideoxy-6-(R)-deuterio-a-D-
compound 6S-2H-5 as colorless oil (6.8 mg, 0.027 mmol, 41%),
glucopyranoside diacetate (6R-2H-2)
whose non-deuteriated isotopomer was previously reported as the
The diazide 17 (25.0 mg, 0.059 mmol) was subjected to hydro-
genolysis and purification as described for 15 to give the title
compound, whose non-deuteriated isotopomer was previously
reported as the HCl salt [31,32], as its diacetate salt in the form of a
free base [30]. ½a D23
ꢀ
¼ ꢁ32.1 (c ¼ 0.1, water). 1H NMR (600 MHz,
a
D2O):
d
¼ 4.49 (d, J1-2 ¼ 8.6 Hz, 1H, H1); 3.57 (d, J6-5 ¼ 5.5 Hz, 1H,
H5); 3.51 (dd, J3-2 ¼ 10.6 Hz, J3-4 ¼ 9.2 Hz, 1H, H3); 3.42 (s, 3H,
-OCH3); 3.34 (dd, J5-4 ¼ 9.4 Hz, J5-6 ¼ 5.5 Hz, 1H, H5); 3.30 (t, J4-
colorless oil (12.3 mg, 0.039 mmol, 67%). ½a D23
¼ þ51.3 (c ¼ 0.5,
ꢀ
¼ J4-5 ¼ 9.4 Hz, 1H, H4); 2.86 (dd, J2-1 ¼ 8.6, J2-3 ¼ 10.6 Hz, 1H, H2);
water). 1H NMR (600 MHz, D2O):
d
¼ 5.01 (d, J1-2 ¼ 3.0 Hz, 1H, H1);
3
1.91 (s, 3H, CH3C(O)-). 13C NMR (150 MHz, D2O):
d
¼ 176.90 (-C(O)
3.84 (d, J5-4 ¼ 9.5 Hz, 1H, H5); 3.81 (t, J3-2 ¼ J3-4 ¼ 10.0 Hz, 1H, H3);
3.41 (s, 1H, H6); 3.41 (s, 3H, -OCH3); 3.36 (t, J4-3 ¼ J4-5 ¼ 9.8 Hz, 1H,
H4); 3.32 (dd, J2-1 ¼ 3.0 Hz, J2-3 ¼ 10.1 Hz, 1H, H2); 1.91 (s, 6H,
OH); 99.75 (C1); 76.02 (C5); 71.90 (C3); 69.66 (C4); 59.99 (t, JC-
¼ 24.0 Hz, C6); 57.34 (-OCH3); 55.64 (C2); 20.52 (CH3C(O)OH).
D
ESI-HRMS: m/z calcd for C7H15DNO5 [MþH]þ 195.1091, found
CH3COOH). 13C NMR (150 MHz, D2O):
d
¼ 179.91 (-C¼O-); 96.16
195.1083.
(C1); 71.06 (C4); 69.36 (C3); 67.67 (C5); 55.49 (-OCH3); 53.59 (C2);

16
T. Kato et al. / Carbohydrate Research 448 (2017) 10e17
39.87 (t, JC-D ¼ 23.6 Hz, C6); 22.32 (CH3COOH). ESI-HRMS: m/z calcd
for C7H16DN2O4 [MþH]þ 194.1251, found 194.1248.
194.1259.
4.1.18. Methyl 6-azido-2,6-dideoxy-2-NH-benzyloxycarbonyl-b-D-
glucopyranoside (21)
4.1.15. Methyl 2-azido-2-deoxy-3,4-di-O-benzyl-6-O-
methanesulfonyl-6-(S)-deuterio-
A solution of 15 (50.0 mg, 0.125 mmol) in dichloromethane
(1 mL) was stirred with N,N-diisopropylethylamine (108 L,
0.625 mmol) and methanesulfonyl chloride (20 L, 0.250 mmol) at
b-D-glucopyranoside (18)
Methyl 2-NH-Cbz glucosaminide 20 [12] (40 mg, 0.12 mmol)
was dissolved in pyridine (0.5 mL) and treated with 2,4,6-
triisopropylbenzenesulfonyl chloride (145 mg, 0.49 mmol) at
ambient temperature and with monitoring by TLC (chlor-
oform:methanol 10:1, Rf ¼ 0.4). After 9 h, the reaction was
quenched with methanol (0.5 mL) and concentrated under reduced
pressure. The residue was purified by silica gel column chroma-
tography (chloroform: methanol 20:1 to 5:1) to give the 6-O-trisyl
derivative (50 mg, 0.083 mmol, 68%), which was dissolved in N,N-
dimethylformamide (1.5 mL), treated with sodium azide (65 mg,
0.99 mmol), and heated to 80 ꢂC with stirring for 6 h and moni-
toring by TLC (chloroform:methanol 10:1, Rf ¼ 0.7). The solvent was
removed under reduced pressure, and the residue purified by silica
gel column chromatography (chloroform: methanol 20:1 to 5:1) to
give the title compound 21 (22.8 mg, 0.065 mmol, 78%) as a
b
m
m
ambient temperature for 2 h, with monitoring by TLC (hexane:ethyl
acetate 2:1, Rf ¼ 0.40). Methanol (0.5 mL) was added, and the
mixture was concentrated under reduced pressure. The residue was
diluted with ethyl acetate and washed with 1 N aqueous hydro-
chloric acid, saturated sodium bicarbonate, and brine. The organic
layer was dried, concentrated, and purified by column chroma-
tography (hexane:ethyl acetate 4:1 to 2:1) to give 18 (52.0 mg,
0.109 mmol, 87%) as a colorless oil. ½a D23
ꢀ
¼ ꢁ29.3 (c ¼ 0.3). 1H NMR
(600 MHz, CDCl3):
d
¼ 7.40e7.26 (m, 10H, arom); 4.92 (d, JCH2a-
¼ 11.0 Hz, 1H, -CH2aPh); 4.88 (d, JCH2a’-CH2b’ ¼ 10.6 Hz, 1H,
CH2b
-CH2a’Ph); 4.82 (d, JCH2b-CH2a ¼ 11.0 Hz, 1H, -CH2bPh); 4.64 (d, JCH2b’-
colorless oil. ½a D23
ꢀ
¼ ꢁ32.9 (c ¼ 0.5 methanol). 1H NMR (600 MHz,
¼ 10.6 Hz, 1H, -CH2b’Ph); 4.32 (d, J6-5 ¼ 4.4 Hz, 1H, H6); 4.20 (d,
CH2a’
J1-2 ¼ 8.1 Hz, 1H, H1); 3.56 (s, 3H, -OCH3); 3.53 (dd, J4-3 ¼ 8.2 Hz, J4-
CD3OD):
d
¼ 7.48e7.28 (m, 5H, arom); 5.04 (s, 2H, -CH2Ph); 4.32 (d,
¼ 9.7 Hz, 1H, H4); 3.50 (dd, J5-4 ¼ 9.7 Hz, J5-6 ¼ 4.4 Hz, 1H, H5);
5
J1-2 ¼ 8.7 Hz, 1H, H1); 3.65 (dd, J6a-5 ¼ 1.9 Hz, J6a-6b ¼ 12.1 Hz, 1H,
3.46 (dd, J3-2 ¼ 9.7 Hz, J 3-4 ¼ 8.3 Hz, 1H, H3); 3.38 (dd, J2-1 ¼ 8.1 Hz,
J2-3 ¼ 9.7 Hz, 1H, H2); 3.02 (s, 3H, -SO2CH3). 13C NMR (150 MHz,
H6a); 3.53 (dd, J6b-5 ¼ 5.6 Hz, J6b-6a ¼ 11.9 Hz, 1H, H6b); 3.53 (s, 3H,
-OCH3); 3.41 (br.t, J3-2 ¼ J
¼ 8.9 Hz, 1H, H3); 3.32 (m, 1H, H2);
3-4
CDCl3):
d
¼ 137.56, 137.25 (2C, arom); 128.83e127.80 (10C, arom);
3.31 (m, 1H, H4); 3.30 (m, 1H, H5). 13C NMR (150 MHz, CD3OD):
102.91 (C1); 82.84 (C3); 76.79 (C4); 75.64 (-CH2-); 76.23 (-CH2’-);
72.88 (C5); 67.27 (t, JC-D ¼ 21.9 Hz, C6); 66.17 (C2); 57.34 (-OCH3);
37.70 (-SO2CH3). ESI-HRMS: m/z calcd for C22H26DN3O7SNa
[M þ Na]þ 501.1530, found 501.1526.
d
¼ 160.33 (-C¼O-); 138.80, 129.82, 129.31 (6C, arom.); 104.47 (C1);
78.02 (C5); 76.53 (C3); 72.60 (C4); 69.48 (-CH2-); 60.71 (-OCH3);
57.59 (C2); 54.31 (C6). ESI-HRMS: m/z calcd for C15H20N4O6Na
[M þ Na]þ 375.1281, found 375.1292.
4.1.16. Methyl 2,6-diazido-2,6-dideoxy-3,4-di-O-benzyl-6-(R)-
deuterio-b-D-glucopyranoside (19)
4.1.19. Methyl 2,6-diamino-2,6-dideoxy-b-D-glucopyranoside
diacetate (3)
A stirred solution of 18 (50.0 mg, 0.105 mmol) in N,N-dime-
thylformamide:water (1.0 mL) was heated at 85 ꢂC for 14 h with
sodium azide (81.9 mg, 1.26 mmol), with monitoring by TLC (hex-
ane:ethyl acetate 3:1, Rf ¼ 0.45). The mixture was diluted with ethyl
acetate and was washed with water. The aqueous layer was
extracted with ethyl acetate and the combined organic layer was
washed with brine, dried, concentrated, and purified by column
chromatography (hexane:ethyl acetate 6:1 to 3:1) to give 19
A solution of 21 (15 mg, 0.046 mmol) in a mixture of 1,4-dioxane
(0.8 mL) and 10% v/v aqueous acetic acid (0.1 mL) was treated with
10% palladium hydroxide on activated charcoal (15 mg), then stir-
red
for
12
h
with
monitoring
by
TLC
(chlor-
oform:methanol:ammonium hydroxide 3:1:0.7, Rf ¼ 0.40) under a
hydrogen atmosphere (45 psi). The reaction mixture was filtered
through Celite®, the filter pad washed with water, and the com-
bined filtrate concentrated and dried under reduced pressure to
give 23 as the diacetate salt in the form of a colorless oil (12.6 mg,
(36.0 mg, 0.085 mmol, 81%) as a colorless oil ½a D23
¼ ꢁ41.0 (c ¼ 0.2).
ꢀ
1H NMR (600 MHz, CDCl3):
d
¼ 7.39e7.23 (m, 10H, arom); 4.91 (d,
0.040 mmol, 88%). ½a D23
ꢀ
¼ ꢁ30.1 (c ¼ 0.3 water). 1H NMR (600 MHz,
JCH2a-CH2b ¼ 10.6 Hz, 1H, -CH2aPh); 4.87 (d, JCH2a’-CH2b’ ¼ 11.0 Hz, 1H,
D2O):
d
¼ 4.59 (d, J1-2 ¼ 8.8 Hz, 1H, H1); 3.65 (dt, J5-4 ¼ 9.5 Hz, J5-
-CH2a’Ph); 4.80 (d, JCH2b-CH2a ¼ 10.6 Hz, 1H, -CH2bPh); 4.58 (d, JCH2b’-
¼ 2.9 Hz, J5e6b ¼ 9.2 Hz, 1H, H5); 3.60 (dd, J3-2 ¼ 10.6 Hz, J3-
6a
¼ 11.0 Hz, 1H, -CH2b’Ph); 4.22 (d, J1-2 ¼ 7.7 Hz, 1H, H1); 3.58 (s,
CH2a’
¼ 9.2 Hz, 1H, H3); 3.52 (s, 3H, -OCH3); 3.44 (dd, J6a-5 ¼ 2.9 Hz, J6a-
¼ 13.1 Hz, 1H, H6a); 3.34 (t, J4-3 ¼ J4-5 ¼ 9.4 Hz, 1H, H4); 3.12 (dd,
4
3H, -OCH3); 3.49e3.39 (m, 4H, H2, H3, H4 and H5); 3.39 (br s, 1H,
6b
H6). 13C NMR (150 MHz, CDCl3):
d
¼ 137.65, 137.41 (2C, arom);
J6b-5 ¼ 9.2 Hz, J6b-6a ¼ 13.1 Hz, 1H, H6b); 2.97 (dd, J2-1 ¼ 8.8 Hz, J2-
129.06e127.57 (10C, arom); 102.75 (C1); 82.96 (C3); 78.24 (C4);
75.61 (-CH2-); 75.19 (-CH2’-); 74.82 (C5); 66.29 (C2); 57.58 (-OCH3);
¼ 10.6 Hz, 1H, H2); 1.84 (s, 3H, CH3OOH). 13C NMR (150 MHz,
3
D2O):
d
¼ 181.21 (CH3COOH); 100.10 (C1); 71.81 (C5); 71.81 (C3);
50.83 (t, JC-D
C
¼
21.3 Hz, C6). ESI-HRMS: m/z calcd for
71.34 (C4); 57.39 (-OCH3); 55.41 (C2); 40.19 (C6); 23.11 (CH3COOH).
ESI-HRMS: m/z calcd for C7H17N2O4 [M þ Na]þ 193.1188, found
193.1197.
21H23DN6O4Na [M þ Na]þ 448.1819, found 448.1815.
4.1.17. Methyl 2,6-diamino-2,6-dideoxy-6-(R)-deuterio-b-D-
glucopyranoside diacetate (6R-2H-3)
Acknowledgement
Hydrogenolysis of 19 (36.0 mg, 0.085 mmol) and purification as
described for 15
a gave the title compound as a colorless oil
We thank the NIH (AI 123352) for support of this work, and
acknowledge support from the NSF (MRI-084043) for the purchase
of the 600 MHz NMR spectrometer in the Lumigen Instrument
Center at Wayne State University.
(19.1 mg, 0.061 mmol, 72%). ½a D23
ꢀ
¼ ꢁ28.6 (c ¼ 0.3, water). 1H NMR
(600 MHz, D2O):
d
¼ 4.62 (d, J1-2 ¼ 8.8 Hz, 1H, H1); 3.65 (dd, J5-
¼ 9.4 Hz, J5-6 ¼ 2.7, 1H, H5); 3.63 (dd, J3-2 ¼ 10.6 Hz, J3-4 ¼ 9.2 Hz,
4
1H, H3); 3.52 (s, 3H, -OCH3); 3.42 (d, J6-5 ¼ 2.8 Hz 1H, H6); 3.35 (t, J4-
¼ J4-5 ¼ 9.3 Hz, 1H, H4); 3.01 (dd, J2-1 ¼ 8.7 Hz, J2-3 ¼ 10.6 Hz, 1H,
3
H2); 1.87 (s, 6H, CH3COOH). 13C NMR (150 MHz, D2O):
d
¼ 180.31
References
(-C¼O-); 99.76 (C1); 71.74 (C5); 71.48 (C3); 71.31 (C4); 57.40
(-OCH3); 55.34 (C2); 39.89 (t, JC-D ¼ 21.3 Hz, C6); 22.32 (CH3COOH).
ESI-HRMS: m/z calcd for C7H16DN2O4 [MþH]þ 194.1251, found
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