114
D. Hernandez Armada et al./Carbohydrate Research 417 (2015) 109–116
of galactose over 5 min, and reacted for 70 min. The mixture was
concentrated to a final volume of approximately 0.5 mL and loaded
4.6. Octyl- -galactopyranoside (5)
D
onto a silica gel column and eluted with 25% CH
fractions containing the product were concentrated in vacuo to afford
3
OH in DCM. The
Compound 4 (35 mg, 0.10 mmol) was dissolved in 1 mL of DMF,
and octanol (150 μL, 0.124 mmol) was added to the solution.
N-Bromosuccinimide (36 mg, 0.2 mmol) was then added to the so-
lution, and after 25 min, Amberlite resin (OH ) was added to quench
the reaction and the solution was stirred until the yellow color dis-
a yellow oil in 60% yield. The product was a complex mixture of di-,
1
–
tri-, and tetra-DVS substituted galactose. H NMR (500 MHz, CDCl
3
):
), 6.24–6.14 (m,
), 5.14–4.91 (m, H-1α), 4.47–3.18 (m, H-1β, H-2, H-3, H-4,
δ = 6.91–6.71 (m, CH
CH CH
H-5, H-6, H-6a, CH
=
CH
2
), 6.52–6.42 (m, CH
=CH
2
=
2
3
appeared. The resin was filtered and washed with CH OH, and the
2
—SO
2
, CH
2
-O). Masses of the di-substituted
solvent was removed under reduced pressure. The residue was pu-
rified by flash chromatography to afford a white solid (18 mg,
+
(C H O
14 24
C
10
S
2
Na M + Na ) calcd. 439.0811 found 439.1, tri-substituted
+
1
(
18
H
32
O
13
S
3
Na M + Na ) calcd. 575.1005, found 575.1 (C22
H
40
O
16
S
4
0.062 mmol) in 62% isolated yield. (α:β, 1:1.1). H NMR (500 MHz,
+
3
M + Na ) calcd.: 711.1199, found: 711.1.
CD
J
OCH
OCH
3
OD): δ = 4.79 (d, 1H;
J
H-1,H-2 = 3.3 Hz, H-1α), 4.19 (d, 1H;
3
H-1,H-2 = 7.6 Hz, H-1β), 3.91–3.65 (m, 11H, H-2α, H-3, H-4, H-6, H-6a,
), 3.56–3.39 (m, 5H, H-2β, H-5, OCH ) 1.69–1.54 (m, 4H;
CH (CH CH ), 0.89 (t,
OD):
2
2
4
.3. Coupling of DVS with N-acetyl-
D
-glucosamine (2)
2
3
CH
(CH ) CH
2 2 5 3
), 1.43–1.23 (m, 20H; OCH
2
2
)
2 5
3
13
6
H; J = 7.0 Hz; O(CH
2
)
7
CH
3
) ppm. C NMR (125 MHz, CD
3
N-Acetyl-d-glucosamine (40 mg, 0.18 mmol) was dissolved in
δ = 105.0, 100.3, 76.6, 75.1, 72.6, 72.4, 71.6, 71.1, 70.9, 70.3, 69.2, 62.8,
1
mL of carbonate buffer (pH 11) and stirred until dissolved. In a
62.5, 33.0, 30.9, 30.6(2), 30.6(0), 30.4, 27.4, 27.2, 23.7, 14.4 ppm. ESI-
+
glass vial, 4.47 mg (0.036 mmol) of divinyl sulfone was dissolved
in 500 μL of carbonate buffer and then added to the glucosamine
solution drop-wise over 5 min, and reacted for an additional 70 min.
The mixture was concentrated to a final volume of approximately
HRMS m/z calcd. for C14
H
28NaO
6
(M + Na ) 315.1778, found 315.1774.
4.7. N′-(β-
D
-lactopyranosyl)-p-toluenesulfono-hydrazide (6)
0
2
.5 mL and loaded onto a Sep-Pak C18 cartridge and eluted with
5% CH OH in H O. The fractions containing the product were con-
The N′-(β-d-lactopyranosyl)-p-toluenesulfono-hydrazide (6) was
3
2
1
5
prepared as previously reported, and obtained in 97% yield.
centrated in vacuo to afford an off-white solid with an isolated yield
1
5
Spectral data match those reported. ESI-HRMS m/z calcd. for
1
of 57%. H NMR (400 MHz, CD
3
OD): δ = 6.98–6.78 (m, CH
), 6.24–6.10 (m, CH CH ), 4.88–4.84 (m, H-1α,
overlaps with residual HOD peak), 4.42 (d, 1H; JH-1,H-2 = 8.4 Hz, H-1β),
.29–4.05 (m, CH —SO , CH —O), 4.02–3.22 (H-3β, H-4, H-5, H-6,
H-6a), 3.93–3.87 (m, H-2α, from COSY), 3.68 (m, H-2β, from COSY),
2
=CH ),
+
C
19
H
30NaO12S (M + Na ) 533.1412, found 533.1401.
6.41–6.26 (m, CH
=CH
2
=
2
3
4
2
2
2
4.8. Octyl- -lactopyranoside (7)
D
3
.41 (m, H-3β, from COSY), 2.01–1.96 (m, Ac). ESI HRMS m/z
Compound 6 (51 mg, 0.10 mmol) was dissolved in 1 mL of dry
DMF, and octanol (150 μL, 0.124 mmol) was added to the solution.
N-Bromosuccinimide (36 mg, 0.2 mmol) was added, and after 25
+
(C
12
H
21NO
8
SNa M + Na ) calcd. 362.1, found 362.1.
–
minutes Amberlite resin (OH ) was added to quench the reaction
4.4. 2-Galactopyranosyl-6-hydroxy-4-methoxy-[1,3,5-triazine] and
and the solution was stirred until the yellow color disappeared. The
2,4-digalactopyranosyl-6-methoxy-[1,3,5-triazine] (3)
3
resin was filtered and washed with CH OH, and the solvent was
removed under reduced pressure. The residue was purified by flash
d-Galactose (80 mg, 0.44 mmol) was dissolved in 6 mL of water,
followed by addition of NaOH (26.4 mg, 0.66 mmol). 4,6-Dichloro-
-methoxy-[1,3,5-triazine] dissolved in 1 mL of acetone (80 mg,
.45 mmol) was added drop wise to the reaction mixture and stirred
chromatography to afford a white solid (14 mg, 0.031 mmol) in 31%
1
isolated yield (α:β, ~2:1). H NMR (500 MHz, CD
3
OD): δ = 4.75 (d,
3
6
0
3
2H, J H-1,H-2 = 3.8 Hz; H-1α, overlaps CD OD), 4.36–4.32 (m, 3H, H-1′),
3
4.27 (d, 1H, J H-1,H-2 = 8.0 Hz; H-1β), 3.90–3.63 (m, 41H, H-2α, H-3,
for 8 h. The mixture was purified by preparative TLC using 12:1:0.4
isopropanol:CH OH:H O as eluent. The product was recovered with
CH OH and dried under reduced pressure to afford a mixture of the
mono and di-substituted products. H NMR (400 MHz, CD
2
H-4, H-5, H-6, H-6a, H-2′, H-3′, H-4′, H-5′, H-6′, H-6a′, OCH ), 3.23
3
3
3
2
(apparent t, 1H, J H-1,H-2 = J H-2,H-3 = 8.0 Hz; H-2α, overlaps CD
3
OD),
3
1.68–1.55 (m, 6H; OCH
OCH CH (CH CH
NMR (125 MHz, CD
2
CH
2
(CH
2
)
5
CH
3
), 1.43–1.23 (m, 30H;
1
3
13
3
OD):
2
2
2
)
5
3
), 0.89 (t, 9H; J = 6.8 Hz; O(CH
OD): δ = 105.1, 99.9, 81.1, 77.1, 74.9, 73.5, 73.3,
2 7
) CH
3
) ppm.
C
δ = 5.23–5.07 (m, H-1α), 4.64–3.39 (m, H-1β, H-2, H-3, H-4, H-5, H-6,
3
+
H-6a, OCH
3
). Masses of the mono-substituted (C10
H
25
15
O
8
N
3
Na ) calcd.
72.6, 72.1, 70.3, 69.4, 62.5, 33.1, 30.6, 30.4, 27.4, 23.7, 14.4 ppm. ESI-
+
+
3
4
28.0757, found 328.0754, di-substituted (C16
90.1285, found 490.1283
H
O
13
N
3
Na ) calcd.
HRMS m/z calcd. for C20
H
38NaO11 (M + Na ) 477.2306, found 477.2306.
4
.9. N′-(2-Acetamido-2-deoxy-β-
D
-glucopyranosyl)-p-
toluenesulfono-hydrazide (8)
4.5. N′-(β-
D
-Galactopyranosyl)-p-toluenesulfono-hydrazide (4)
Compound 8 (1.24 g, 2.4 mmol) was prepared as previously
Compound 4 (1.95 g, 5.6 mmol) was prepared as previously
9
reported, with an isolated yield of 99%. Spectral data match those
1
5
reported. Isolated yield was 99%, and only the β-anomer was ob-
9
reported, and only the β-anomer was observed by NMR. ESI-
1
served. H NMR (500 MHz, CD
3
OD): δ = 7.79 (m, 2H, Ar), 7.37 (m,
+
HRMS m/z calcd. for C15
12.1147.
H
23
N
3
NaO
7
S (M + Na ) 412.1149, found
3
3
2
H, Ar), 3.75 (dd, 1H; JH-4,H-5 = 1.0 Hz, JH-3,H-4 = 3.5 Hz, H-4), 3.71 (dd,
4
3
J
H-5,H-6 = 7.6 Hz, 3JH-6,H-6a = 11.4 Hz, H-6), 3.71 (dd, 1H;
1
H;
3
H-5,H-6a = 4.4 Hz, 3
J
J
H-6,H-6a = 11.4 Hz, H-6a), 3.62–3.55 (m, 2H, H-1
[
3.61 ppm from HSQC], H-2 [3.58 ppm from HSQC]), 3.40 (dd, 1H;
4.10. Octyl-2-acetamido-2-deoxy-β- -glucopyranoside (9)
D
3
3
3
J
H-3,H-4 = 3.5 Hz, JH-2,H-3 = 9.0 Hz, H-3), 3.71 (ddd, 1H; JH-4,H-5 = 1.0 Hz,
3
3
J
H-5,H-6a = 4.4 Hz, JH-5,H-6 = 7.6 Hz, H-5), 2.42 (apparent s, 3H, PhCH
3
)
Compound 9 was prepared as previously reported, and iso-
9
ppm. A starting material impurity was detected at δ = 7.53, 7.22, and
lated as a white solid in 70% yield. Spectral data match those
13
9
2
9
C
.34 ppm. C NMR (125 MHz, CD
3
OD): δ = 145.1, 137.4, 130.6, 129.1,
reported, and only the β-anomer was observed by NMR. ESI-
+
2.3, 78.0, 75.2, 70.6, 69.4, 63.0, 21.5 ppm. ESI-HRMS m/z calcd. for
HRMS m/z calcd. for C16
356.2044.
H
31NO
6
Na (M + Na ) 356.2044, found
+
+
13
2 20 7
H N O SNa (M + Na ) 371.0883, found 371.0878.