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Bioconjugate Chemistry
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C NMR (101 MHz, CDCl ) δ 170.27, 169.75 and 169.58
andH2O (1 mL) was added to quench the reaction. After removal
of the solvent, the residue was suspended in brine and extracted
three times with ethyl acetate. The combined organic layers were
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(C(O)CH3), 88.19 (Cꢀ1), 71.29 (Cꢀ5), 69.47 (Cꢀ2), 67.05 (Cꢀ4),
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0.95 (Cꢀ6), 58.32 (Cꢀ3), 20.69, 20.60, 20.50 (C(O)CH3).
Preparation of compound 4 (Triꢀisopropylsilyl 3ꢀazidoꢀ2,4,6ꢀ
triꢀOꢀacetylꢀ1ꢀthioꢀβꢀ ꢀgalactopyranoside). To a solution of 3
770 mg, 1.95 mmol) in dry CH CN (10 mL) N gas was purged
dried over Na SO before the solvent was removed. The resulting
2 4
D
oil was purified by silica chromatography (hexanes: EtOAc = 1: 1
(
→ 1: 2) to give compound 7 (120 mg, 39 %) as a colorless oil.
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for 10 mins, then K CO (809 mg, 5.86 mmol) was added folꢀ
H NMR (400 MHz, Chloroformꢀd) δ 4.17 (d, J = 2.4 Hz, 2H),
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lowed by triꢀisoꢀpropylsilylthiol (TIPSSH, 628 ꢁL, 2.93 mmol),
and the reaction was stirred for 3 h at room temperature. After
complete conversion of the starting material according to TLC
monitoring, the solvent was evaporated and the residue was disꢀ
solved in CH Cl (20 mL), washed with H O (2 x 20 mL). The
3.73 – 3.53 (m, 18H), 2.46 (t, J = 6.6 Hz, 2H), 2.40 (t, J = 2.4 Hz,
1H), 1.41 (s, 9H).
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C NMR (101 MHz, Chloroformꢀd) δ 170.82 , 80.42 , 74.43 ,
70.57 , 70.56 , 70.53 , 70.46 , 70.36 , 70.33 , 69.07 , 69.07 , 66.85
, 58.34 , 36.24 , 28.08 , 28.05 .
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organic layer was dried over Na SO , filtered and concentrated in
vacuo. Silica chromatography (hexanes: EtOAc = 4: 1) yielded 4
Preparation of compound 8. The compound 7 (33 mg, 0.092
mmol) and compound 6 crude (48 mg) were dissolved into
CH3CN (2.0 mL) and then CuI (18 mg, 0.093 mmol) was added
into the solution. The resulting mixture was heated under microꢀ
wave irradiation to 80 °C for 90 mins. After complete conversion
of the starting material according to TLC monitoring, the mixture
2
4
as a white solid (300 mg, 30%).
1
H NMR (400 MHz, CDCl3) δ 5.42 (dd, J = 3.5, 1.0 Hz, 1H, H4),
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.20 (t, J = 9.8 Hz, 1H, H2), 4.60 (d, J = 9.5 Hz, 1H, H1), 4.11
(dd, J = 11.5, 5.6 Hz, 1H, H6a), 4.00 (dd, J = 11.6, 7.1 Hz, 1H,
H6b), 3.79 (ddd, J = 6.9, 5.5, 1.1 Hz, 1H, H5), 3.53 (dd, J = 10.1,
was concentrated in vacuo, and then H O (1.0 mL) was added. A
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.4 Hz, 1H, H3), 2.15, 2.12, 2.02 (3S, 9H, 3 COCH3), 1.25 (m,
H, ꢀSiC3H3), 1.14 – 1.07 (m, 18H, ꢀSiC3H3C6H18).
clear solution was obtained after centrifuge, which was purified
by sizeꢀexclusion chromatography (BioꢀGel P2 fine; column 2.5
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C NMR (101 MHz, CDCl ) δ 170.57, 170.25, 169.39 (3
cm × 120 cm; flow rate 0.3 ml/min; elution with H O/nꢀButanol =
3
2
COCH ), 80.37 (Cꢀ1), 75.47 (Cꢀ5), 62.87 (Cꢀ3), 72.17 (Cꢀ2),
95/5). The fractions containing the product were pooled and
freezeꢀdried to give compound 8 (52 mg, 0.060 mmol, 64 %) as a
white fluffy solid.
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8.12 (Cꢀ4), 62.13 (Cꢀ6), 20.84, 20.76, 20.51 (3COCH ), 18.52,
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18.22 (6 –SiCHCH ), 12.76 (3 –SiCH).
HRMS (EI, m/z): calculated for C21H37N3O7SSiNa+
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H NMR (500 MHz, D2O) δ 8.55 (s, 1H), 8.26 (s, 1H), 7.87 (d, J
(
[M+Na]+): 526.2014, found 526.2011.
Preparation of compound 5 (3ꢀazidoꢀ3’ꢀphenylꢀ2,2’,4,4’,6,6’ꢀ
hexaꢀOꢀacetyl βꢀ ꢀthiodigalactoside). The solution of 2 (190
mg, 0.38 mmol) in dry CH CN (10 mL) was added by 4 (193 mg,
= 7.7 Hz, 2H), 7.55 (t, J = 7.6 Hz, 2H), 7.48 (t, J = 7.5 Hz, 1H),
6.81 – 6.69 (m, 2H), 5.14 (dd, J = 9.8, 7.4 Hz, 2H), 5.02 (td, J =
13.7, 10.5, 2.9 Hz, 2H), 4.80 – 4.73 (m, 2H), 4.74 (s, 2H), 4.46
(dt, J = 27.5, 10.3 Hz, 3H), 4.26 (dd, J = 22.4, 3.0 Hz, 2H), 4.06
(td, J = 8.5, 4.4 Hz, 3H), 3.89 – 3.76 (m, 4H), 3.75 (t, J = 2.4 Hz,
2H), 3.66 (dq, J = 7.8, 4.3, 3.5 Hz, 16H), 3.40 – 3.26 (m, 2H),
3.00 – 2.88 (m, 4H), 2.56 (t, J = 6.0 Hz, 2H), 1.45 (s, 9H).
D
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.38 mmol), N2 gas was purged for 10 mins through the solution,
and tetraꢀnꢀbutylammoniumfluoride (TBAF, 1M in THF, 460 ꢁL)
was added. Following complete conversion of the starting materiꢀ
al after 5 mins according to TLC analysis, the solvent was evapoꢀ
rated and silica chromatography (hexanes:EtOAc = 1: 1 → 1: 2)
gave compound 5 (160 mg, 54 %) as white solid.
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C NMR (126 MHz, D2O, extracted from HSQC) δ 121.43,
124.35, 125.65, 126.36, 125.65, 129.20, 129.23, 128.75, 84.13,
84.12, 66.86, 66.97, 63.11, 66.85, 67.89, 79.54, 79.55, 61.03,
66.44, 61.05, 61.06, 69.53, 69.52, 69.50, 70.21, 42.99, 42.99,
35.06, 27.18, 27.17, 27.18.
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H NMR (400 MHz, CDCl3) δ = 7.80 (s, 1H, triazole), 7.77ꢀ7.71
(
(
m, 2H, ar), 7.43ꢀ7.28 (m, 3H, ar), 5.75ꢀ5.68 (m, 1H, Hꢀ2’), 5.61
d, 1H, J =3.2 Hz, Hꢀ4’), 5.47 (d,1H, J4,3 =3.4 Hz, J4,5 = 1.1 Hz,
+
HRMS (EI, m/z) calculated for C38H58N6O15SNa+ ([M+Na] ):
Hꢀ4), 5.21 (dd, 1H, J3’,2’= 8.6 Hz, J3’,4’= 2.5 Hz, Hꢀ3’), 5.17 (m,
1H, Hꢀ2), 4.98 (1H, d, J1’,2’= 9.8 Hz, Hꢀ1’), 4.84 (1H, J1,2 =
893.3579, found 893.3595.
Preparation of compound 9. Compound 8 (52 mg, 0.060 mmol)
was added into TFA/CH Cl (10 mL, 1:1) and the solution was
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0.0 Hz, Hꢀ1), 4.11 (m, 5H, Hꢀ5’, Hꢀ6ab, Hꢀ6a’b’), 3.89 (1H, td,
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J5,4 = 1.2 Hz, J5,6ab = 6.4 Hz, Hꢀ5), 3.67 (1H, J3,2 = 10.1 Hz,
J3,4 = 3.4 Hz, Hꢀ3), 2.15, 2.13, 2.08, 2.05, 2.04, and 2.02 (6s,
stirred for 2 h at room temperature. After being fully evaporated,
the residue was purified by sizeꢀexclusion chromatography (Bioꢀ
Gel P2 fine; column 2.5 cm × 120 cm; flow rate 0.3 ml/min;
total 18H, C(O)CH ).
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C NMR (101 MHz, Chloroformꢀd) δ 170.34 , 170.25 , 169.86 ,
elution with H O/nꢀButanol = 95/5). The fractions containing the
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169.54 , 169.37 , 168.70 , 147.92 , 129.96 , 128.90 , 128.47 ,
125.68 , 118.26 , 82.11 , 81.51 , 77.34 , 77.02 , 76.70 , 75.53,
product were pooled and freezeꢀdried to give compound 9 (48 mg,
0.059 mmol, 98 %) as a white solid.
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,
8.71 , 68.43 , 67.66 , 66.33 , 62.94 , 62.78 , 61.56 , 61.38 , 20.76
20.68 , 20.62 , 20.61 , 20.47 , 20.38 .
H NMR (400 MHz, Deuterium Oxide) δ 8.40 (s, 1H), 8.11 (s,
1H), 7.75 – 7.68 (m, 2H), 7.44 – 7.37 (m, 2H), 7.36 – 7.28 (m,
1H), 4.99 (dd, J = 9.8, 6.1 Hz, 2H), 4.87 (ddd, J = 11.9, 10.7, 3.0
Hz, 2H), 4.59 (s, 2H), 4.31 (dt, J = 20.7, 10.2 Hz, 2H), 4.16 – 4.06
(m, 2H), 3.91 (td, J = 7.6, 4.4 Hz, 2H), 3.74 – 3.60 (m, 5H), 3.60 –
3.48 (m, 17H), 2.31 (t, J = 6.7 Hz, 2H).
HRMS (EI, m/z) calculated for C32H38N6O14SH+ ([M+H]+):
63.2239, found 763.2277.
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Preparation of compound 6 (3ꢀ(4ꢀphenylꢀ1Hꢀ1,2,3ꢀtriazol)ꢀ3’ꢀ
azidoꢀthiodigalactoside). NaOMe (40 mg, 2.5 mmol) was added
in the solution of compound 5 (120 mg, 0.16 mmol) in CH OH
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C NMR (101 MHz, Deuterium Oxide) δ 180.11, 163.09 , 162.74
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(5.0 mL) and the mixture was stirred for 6 h at room temperature.
, 147.43 , 143.78 , 129.19 , 128.82 , 125.65 , 124.36 , 121.38 ,
117.70 , 114.80 , 84.10 , 79.48 , 69.48 , 69.45 , 69.43 , 69.17 ,
68.85 , 67.98 , 67.93 , 67.84 , 66.94 , 66.83 , 66.77 , 66.74 , 63.04
, 61.00 , 37.52 .
The solution was neutralized with DOWEXꢀH+ resin, filtered,
and evaporated. Crude 6 was obtained as a white solid and used in
next step without further purification.
+
+
Preparation of compound 7. To
a
solution 3,6,9,12ꢀ
HRMS (EI, m/z): calculated for [C34H50N6O15S+H] ([M+H] ):
815.3128, found 815.3135.
tetraoxapentadecꢀ14ꢀynꢀ1ꢀol of (200 mg, 0.86 mmol) in 5 mL of
THF was added sodium (0.6 mg,0.025 mmol). When the sodium
was dissolved, tertꢀbutyl acrylate (0.125 mL, 0.86 mmol) was
added. The solution was stirred for 20h at room temperature
Preparation of compound 10. Compound 9 (22 mg, 0.027
mmol) was dissolved in anhydrous DMF (0.5 mL) and DiPEA
(4.69 µl, 0.027 mmol) was added, followed by addition of TSTU
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