10.1002/ejoc.201601534
European Journal of Organic Chemistry
FULL PAPER
(CH2Cl2/MeOH, 100/090/10, v/v) to afford the fluorescent glycoside as
an oil (10 mg, 70%). Rf = 0.61 (CH2Cl2/MeOH, 4/1, v/v); αD20.3 = -10 (c =
0.1, MeOH); IR (film) 3035, 2931, 2986, 2040, 1719 cm-1; UV-VIS: (MeOH),
Hydrolytic stability experiment. A series of citric acid-Na2HPO4 buffers
at various pH (pH 3, 4, 5, 6) and buffer concentrations (25, 50, 100, 200
mM), containing 0.1% NaN3, were prepared. To the dansylated glycosides
12 and 13 (in separate flasks), a stock solution of internal standard 16
(0.25 equiv. compared to 12 or 13) in H2O was added and the resulting
solutions were divided into separate reaction vessels and lyophilized to
obtain 2 μmol of dansyl-glycoside 12 or 13 per reaction vessel. To each
reaction vessel, the previously prepared buffers (50-800 µL, depending on
experiment) were added. The hydrolysis reactions were monitored over
time using HPLC-UV analysis (column: BEH300 C18 5 μm, 4.0 x 150 mm,
35% MeCN in H2O). The integrated chromatograms of glycosides 12 or 13
were measured against the integral of the internal standard. A line was
fitted to the data according to the least-squares method, and the half-lives
were calculated based on the fitted line. In control experiments, an excess
of internal standard and NaN3 were added. In addition, reactions were
performed in PBS at various glycan concentrations (12.5, 25, 100, 200 µM).
max = 331 nm; 1H NMR (500 MHz, D2O) δ 8.52 (d, 1H, J2,3 = 8.6 Hz, H-2),
8.33 (d, 1H, J7,8 = 8.8 Hz, H-8), 8.28 (d, 1H, J3,4 = 7.4 Hz, H-4), 7.74 (t, 1H,
J3,4 = J2,3 = 7.7 Hz, H-3), 7.71 (t, 1H, J6,7 = J7,8 = 7.7 Hz, H-7), 7.44 (d, 1H,
J6,7 = 7.7 Hz, H-6), 4.07 (d, 1H, J1′′,2′′ = 9.7 Hz, H-1′), 3.88 (d, 1H, J6a′′,6b′′
=
12.0 Hz, H-6b′′), 3.81 (t, 1H, J1′′,2′′ = J2′′,3′′ = 9.8 Hz, H-2′′), 3.71 (dd, 1H,
J6a′′,5′′ = 5.7 Hz, J6a′′,6b′′ = 12.3 Hz, H-6a′′), 3.51 – 3.40 (m, 3H, NO-CH2-1′,
H-3′′), 3.36 (t, 1H, J4′′,5′′ = J3′′,4′′ = 9.3 Hz, H-4′′), 3.32 – 3.25 (m, 1H, H-5′′),
3.18 – 3.09 (m, 2H, H-2′), 2.88 (s, 6H, N(CH3)2), 2.30 (s, 3H, CH3 Ac), 1.97
(s, 3H, N-CH3); 13C NMR (125 MHz, D2O) δ 173.9 (C=O), 150.7 (C-4a),
134.0 (C-5), 130.1 (C-2), 129.5 (C-4), 128.9 (C-7), 128.8 (C-8a, C-1),
124.0 (C-3), 119.1 (C-8), 116.0 (C-6), 91.4 (C-1′′), 77.3 (C-5′′), 75.2 (C-3′′),
69.8, 69.6 (C-4′′, C-1′), 60.8 (C-6′′), 52.1 (C-2′′), 44.9 (N(CH3)2), 41.3 (C-
2′), 37.6 (CH3 Ac), 22.1 (N-CH3); HRMS(ESI) m/z calcd. for
[C23H35N4O8S]+: 527.2170, obsd.: 527.2178.
5-(Dimethylamino)-N-(3-[(methoxy)(2-acetamido-2-deoxy-β-D-
Acknowledgements
glucopyranosyl)amino]-propyl)-naphthalene-1-sulfonamide (13). To a
solution of amine 12 (52 mg, 0.17 mmol) in H2O (4 mL), NaHCO3 (84 mg,
1.0 mmol) was added. Dansyl chloride (70 mg, 0.26 mmol) was dissolved
in distilled THF (2 mL) and added drop wise to the reaction mixture. After
2 h the THF was concentrated in vacuo and the remaining aqueous
reaction mixture was purified by reverse phase column chromatography
(C18, H2O/MeOH, 100/0 → 25/75, v/v) to give fluorescent glycoside 13 (72
mg, 79%); Rf = 0.34 (CH2Cl2/MeOH, 8/1, v/v); αD17.8 = -8.2 (c = 0.1, MeOH);
IR (film) 3501, 3334, 3242, 3084, 2942, 2873, 2839, 2791, 1645, 1571,
1504, 1454, 1440, 1407, 1376, 1355, 1309, 1231, 1201, 1182, 1140, 1076,
The authors would like to thank the Wellington Medical Research
Foundation for financial support.
Keywords: Carbohydrates • Conjugates • Oxyamine • Linker •
Organic Synthesis
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1060, 1027, 944, 790 cm-1; 1H NMR (500 MHz, D2O) 8.50 (d, 1H, J2,3
=
8.6 Hz, H-2), 8.30 (d, 1H, J3,4 = 8.7 Hz, H-4), 8.28 (d, 1H, J7,8 = 7.3 Hz, H-
8), 7.75 – 7.67 (m, 2H, H-3, H-7), 7.43 (d, 1H, J6,7 = 7.6 Hz, H-6), 4.12 (d,
1H, J1′′,2′′ = 9.8 Hz, H-1′′), 3.84 (dd, 1H, J5′′,6a′′ = 1.4 Hz, J6a′′,6b′′ = 12.0 Hz, H-
6a′′), 3.67 (dd, 1H, J1′′,2′′ = J2′′,3′′ = 9.8 Hz, H-2′′), 3.65 (dd, 1H, J5′′,6b′′ = 5.6
Hz, J6a′′,6b′′ = 12.0 Hz, H-6b′′), 3.41 (dd, 1H, J2′′,3′′ = J3′′,4′′ = 9.6 Hz, H-3′′),
3.31 (t, 1H, J3′′,4′′ = J4′′,5′′ = 9.0 Hz, H-4′′), 3.27 – 3.22 (m, 1H, H-5′′), 3.03 (s,
3H, OCH3), 2.99 – 2.86 (m, 2H, CH2-3′), 2.87 (s, 6H, 2 × N-CH3), 2.70 –
2.59 (m, 2H, CH2-1′), 1.84 (s, 3H, CH3 Ac), 1.48 (p, 2H, J1′,2′ = J2′,3′ = 6.8
Hz, CH2-2′); 13C NMR (125 MHz, D2O) 173.5 (C=O Ac), 150.7, 133.5, (C-
4a, C-8a), 130.1 (C-2), 129.9 (C-8), 128.9, 128.81, 128.77 (C-1, C-5, C3/7),
124.0 (C-3/7), 118.9 (C-4), 116.02 (C-6), 89.9 (C-1′′), 77.3 (C-5′′), 75.3
(C-3′′), 69.5 (C-4′′), 60.8 (C-6′′), 60.6 (OCH3), 52.1 (C-2′′), 48.0 (C-1′), 44.9
(N(CH3)2), 40.4 (C-3′), 26.0 (C-2′), 22.0 (CH3 Ac); HRMS(ESI) m/z calcd.
for [C24H37N4O8S]+: 541.2327, obsd.: 541.2333.
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N-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-N-methyl-O-
methylhydroxylamine (17). N-Acetylglucosamine (1.0 g, 4.5 mmol) and
N,O-dimethylhydroxylamine hydrochloride (1.3 g, 13 mmol) were
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NH4HCO3 was added to the reaction mixture, which was then lyophilised.
The crude material was purified by silica gel column chromatography
(CH2Cl2/MeOH, 85/15, v/v), followed by ion exchange (Dowex-OH-) to
afford the title compound as a white solid (0.77 g, 65%). Rf = 0.26
(MeOH/CH2Cl2, 1:4, v/v); αD20.1 = -10 (c = 0.1, MeOH); IR (film) 3290, 2938,
2895, 1647, 1561, 1035 cm-1; 1H NMR (500 MHz, D2O) δ 4.19 (d, 1H, J1,2
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= 9.6 Hz, H-1), 3.99 (t, 1H, J1,2 = J2,3 = 9.8 Hz, H-2), 3.93 (dd, 1H, J6a,6b
=
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12.3 Hz, J5,6a = 2.2 Hz, H-6a), 3.75 (dd, 1H, J6b,6a = 12.3 Hz, J5,6b = 5.5 Hz,
H-6b), 3.52 (dd, 1H, J2,3 = J3,4 = 10.0 Hz, H-3), 3.46 (s, 3H, OCH3), 3.45 –
3.40 (m, 2H, H-4, H-5), 2.74 (s, 3H, NCH3), 2.05 (s, 3H, CH3 Ac); 13C NMR
(125 MHz, D2O) δ 173.9 (C=O Ac), 91.2 (C-1), 77.4 (C-5), 75.3 (C-3), 69.6
(C-4), 60.8 (C-6), 49.3 (OCH3), 52.1 (C-2), 38.1 (NCH3), 22.2 (CH3 Ac);
HRMS(ESI) m/z calcd. for [C10H21N2O6]+: 265.1394, obsd.: 265.1400.
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