ArH), 5.58 (1H, dt, J 6.56, 3.20, H-1), 5.13–5.06 (4H, m,
2 × CH2Ph), 4.73 (2H, br s, CH2Ph-6), 4.62 (1H, ddd, J 8.70,
9.92, 50.96, H-4), 4.52 (1H, d, J 12.06, CH2Ph-3), 4.47 (1H, d,
J 12.06, CH2Ph-3), 4.15–3.87 (3H, m, H-2, H-3, H-5), 3.66–3.55
(2H, m, H-6a, H-6b), 1.81 (3H, s, COCH3).
3.12 (1H, dt, J 6.09, 9.73, H-3), 1.82 (3H, s, COCH3);
FAB-HRMS, m/z found MϩHϩ 262.1311, C11H20NO6 requires
262.1291.
Allyl 2-acetamido-2-deoxy-4,6-O-benzylidene-ꢀ-D-gluco-
pyranoside 13
Disodium 2-acetamido-2,4-dideoxy-4-fluoro-ꢀ-D-glucopyranosyl
phosphate 11
To a cooled solution (0 ЊC) of 12 (10.4 g, 40 mmol) in DMF
(130 cm3) was added ( )-camphor-10-sulfonic acid (929 mg,
4 mmol) and benzaldehyde dimethyl acetal (7.8 cm3, 52 mmol)
dropwise. The solution was stirred for 12 h at rt, then the
solvent was evaporated under reduced pressure to yield a pale
yellow oil. CHCl3–MeOH (6 : 1) was poured into the flask to
dissolve the crude product, and 400 cm3 water was added to the
flask, and stirred for 5 min. Then 2000 cm3 hexane was added
slowly to precipitate the product 13 (11.3 g, 81%) as a white
To a solution of 10 (35 mg, 0.053 mmol) in EtOH (1.6 cm3) and
10% aq. NaHCO3 (1 cm3) was added 25 mg 5% Pd/C. After
stirring for 1 d at rt under a hydrogen atmosphere, the mixture
was filtrated through a Celite pad. The solvent was evaporated
under reduced pressure to yield the crude product, which was
purified by G-10 gel chromatography (H2O) to give 11 (15 mg,
82%) as a colorless foamy solid (freeze-dried); [α]25 ϩ36.2Њ
D
(c 0.1 in H2O); δH (400 MHz, D2O) 5.35 (1H, dt, J 7.63, 3.20,
H-1), 4.42 (1H, ddd, J 8.70, 9.92, 50.96, H-4), 4.03–4.14 (2H, m,
H-2, H-3), 3.96 (1H, dt, J 9.92, 1.98, H-5), 3.87 (1H, dt, J 12.66,
1.98, H-6a), 3.78 (1H, ddd, J 1.98, 4.27, 12.66, H-6b), 2.05
solid; [α]25 ϩ67.3Њ (c 0.1 in MeOH); mp 208–210 ЊC (EtOH);
D
δ (400 MHz, DMSO-d6) 7.91 (1H, d, J 8.34, NH), 7.46–7.36
(5H, m, ArH), 5.94–5.84 (1H, m, CH᎐C), 5.61 (1H, s, PhCH),
᎐
5.33 (1H, dd, J 1.71, 17.32, C᎐CH ), 5.17 (1H, dd, J 1.71, 10.37,
᎐
2
(3H, s, COCH ); δ (100 MHz, D O) 175.6 (C᎐O), 93.4 (C-1),
C᎐CH ), 5.15 (1H, d, J 6.09, OH-3), 4.75 (1H, d, J 3.74, H-1),
᎐
2
᎐
3
C
2
90.1 (d, J 179.3, C-4), 70.7 (C-3), 70.3 (C-5), 60.9 (C-6), 54.5
(C-2), 22.9 (CH3); FAB-HRMS, m/z found MϩHϩ 348.0215,
C8H14NO8Na2PF requires 348.0236.
4.17 –4.11 (2H, m, H-6a, CH -C᎐C), 3.95 (1H, dd, J 6.41, 13.68,
᎐
2
CH -C᎐C), 3.84 (1H, ddd,
J 3.74, 8.34, 10.25, H-2),
᎐
2
3.75–3.61 (3H, m, H-3, H-5, H-6), 3.48 (1H, t, J 9.73, H-4), 1.84
(3H, s, COCH3); FAB-HRMS, m/z found MϩHϩ 350.1589,
C18H24NO6 requires 350.1604.
Disodium uridine 5Ј-(2-acetamido-2,4-dideoxy-4-fluoro-ꢀ-D-
glucopyranosyl) diphosphate 1
Allyl 2-acetamido-2-deoxy-3-O-benzyl-4,6-O-benzylidene-ꢀ-D-
glucopyranoside 14
A reaction mixture (2.4 cm3) containing 50 mM Tris-HCl (pH
7.5), 5 mM UTP-3Na, 5 mM MgCl2, 0.01 cm3 UDP-GlcNAc
pyrophosphorylase (2.4 units) and 11 (4.2 mg, 0.012 mmol) was
incubated at 37 ЊC for 1 h and the reaction was stopped by
boiling for 3 min. Quantitative determination of nucleotides
and bases was carried out by HPLC with a YMC-packed
column Hydrosphere C-18 (4.5 × 150 mm, Eishin Chemical,
Japan) at 30 ЊC with detection at 260 nm. The mobile phase was
0.2 M triethylamine containing phosphoric acid (pH 6.0) and
the flow rate was 0.6 cm3 minϪ1. Further purification was
carried out by G-10 gel chromatography (H2O) to afford pure 1
(1.8 mg, 23%) as a white solid (freeze-dried); [α]25D ϩ47.9Њ (c 0.1
in H2O); δH (600 MHz, D2O) 7.60 (1H, d, J 8.14, uridine-HЉ-6),
5.67 (1H, d, J 8.14, uridine-HЉ-5), 5.66 (1H, d, J 5.00, rib-HЈ-1),
5.21 (1H, dt, J 7.33, 3.28, H-1), 4.18 (1H, ddd, J 7.78, 9.96,
50.58, H-4), 4.06 (1H, t, J 5.00, rib-HЈ-2), 4.01 (1H, t, J 5.00,
rib-HЈ-3), 4.00–3.97 (1H, m, rib-HЈ-4), 3.94 (1H, ddd, J 2.27,
4.77, 11.70, rib-HЈ-5a), 3.86 (1H, ddd, J 4.07, 5.33, 11.70,
rib-HЈ-5b), 3.82–3.73 (3H, m, H-5, H-3, H-2), 3.58–3.47 (2H,
m, H-6a, H-6b), 1.78 (3H, s, COCH3); δC (100 MHz, D2O) 175.6
To a cooled solution (0 ЊC) of 13 (1.74 g, 5 mmol) in DMF
(36 cm3) was added barium oxide (1.53 g, 10 mmol), barium
hydroxide (1.58 g, 5 mmol) and benzyl bromide (1.2 cm3,
10 mmol) dropwise. The solution was stirred for 12 h at rt. After
filtration through a Celite pad, the solvent was evaporated
under reduced pressure to yield a pale yellow oil. CHCl3–
MeOH (6 : 1) was poured into the flask to dissolve the crude
product and then 90 cm3 hexane was added slowly to precipitate
the product 14 (1.66 g, 76%) as a white solid; [α]25 ϩ 41.9Њ
D
(c 0.1 in MeOH); mp 224–225 ЊC (EtOH); δH (400 MHz,
CDCl ) 7.52–7.28 (10H, m, ArH), 5.91–5.81 (1H, m, CH᎐C),
᎐
3
5.60 (1H, s, PhCH), 5.34 (1H, d, J 9.16, NH), 5.28–5.19 (2H, m,
C᎐CH ), 4.92 (1H, d, J 12.36, PhCH ), 4.86 (1H, d, J 3.66,
᎐
2
2
H-1), 4.64 (1H, d, J 12.36, PhCH2), 4.32–4.26 (2H, m, H-2,
H-6a), 4.15 (1H, ddt, J 5.34, 12.97, 1.38, CH -C᎐C), 3.96 (1H,
᎐
2
ddt, J 6.26, 12.97, 1.22, CH -C᎐C), 3.89–3.71 (4H, m, H-3, H-4,
᎐
2
H-5, H-6), 1.91 (3H, s, COCH3); FAB-HRMS, m/z found
MϩHϩ 440.2066, C25H30NO6 requires 440.2073.
(C᎐O), 167.0 (uridine-CЉ-2), 152.6 (uridine-CЉ-4), 142.4
᎐
Allyl 2-acetamido-2-deoxy-3,6-di-O-benzyl-ꢀ-D-glucopyranoside
15
(uridine-CЉ-6), 103.4 (uridine-CЉ-5), 92.2 (C-1), 90.6 (rib-CЈ-1),
90.0 (d, J 180.2, C-4), 83.9 (rib-CЈ-4), 74.6 (rib-CЈ-2), 73.9 (rib-
CЈ-3), 71.4 (C-3), 70.4 (C-5), 65.8 (C-6), 60.5 (rib-CЈ-5), 54.0
(C-2), 22.9 (CH3); ESI-HRMS, m/z found MϪ2NaϩϩHϩ
608.0718, C17H25N3O16P2F requires 608. 0833.
To a solution of 14 (878 mg, 2 mmol) in THF (24 cm3) was
added 4 Å MS (1 g) and NaBH3CN (2 g, 32 mmol), and then
the mixture was stirred for 20 min at rt. Cooled to 0 ЊC, 2.0 M
HCl–diethyl ether solution was added dropwise until the
mixture attained pH 3 and it was then stirred for 1 h at rt. After
filtration through a Celite pad, the solution was diluted with
diethyl ether, washed with H2O, aq. NaHCO3, brine and dried
(MgSO4). The solvent was evaporated under reduced pressure
to yield the product as a pale yellow oil, which was purified by
silica chromatography (25 : 1 chloroform–MeOH) to give 15
(611 mg, 69%) as a white solid; [α]D ϩ30.7Њ (c 0.1 in CHCl3); mp
102–103 ЊC (ethyl acetate–ether); δH (400 MHz, CDCl3) 7.37–
Allyl 2-acetamido-2-deoxy-ꢀ-D-glucopyranoside 12
To a solution of N-acetyl--glucosamine (22.1 g, 0.1 mol) in
allyl alcohol (400 cm3) was added a BF3 diethyl ether complex
(2 cm3, 0.2 mmol), and the mixture was refluxed for 2 h at
130 ЊC. Cooled to rt, the solvent was evaporated under reduced
pressure. EtOH (200 cm3) was added to dissolve the crude
product, then diisopropyl ether was added to the flask to
precipitate the product 12 (23 g, 88%) as a white powder; [α]25
D
ϩ175.8Њ (c 0.1 in MeOH); mp 169–171 ЊC (EtOH) (lit.,8 172–
174 ЊC); δH (400 MHz, DMSO-d6) 7.73 (1H, d, J 8.34, NH),
7.28 (10H, m, ArH), 5.92–5.83 (1H, m, CH᎐C), 5.41 (1H, d,
᎐
J 9.28, NH), 5.28–5.18 (2H, m, C᎐CH), 4.83 (1H, d, J 3.66,
᎐
5.91–5.82 (1H, m, CH᎐C), 5.30 (1H, dd, J 1.71, 17.32, C᎐CH ),
H-1), 4.76 (1H, d, J 11.96, PhCH2-3), 4.70 (1H, d, J 11.96,
PhCH2-6), 4.62 (1H, d, J 11.96, PhCH2-6), 4.55 (1H, d, J 11.96,
᎐
᎐
2
5.13 (1H, dd, J 1.71, 10.37, C᎐CH ), 4.97 (1H, d, J 6.09, OH-3),
᎐
2
4.69 (1H, d, J 5.54, OH-4), 4.66 (1H, d, J 3.74, H-1), 4.49 (1H, t,
PhCH -3), 4.29–4.14 (2H, m, H-2, CH -C᎐C), 3.98–3.93 (1H,
᎐
2 2
J 6.09, OH-6), 4.07 (1H, dd, J 5.24, 13.68, CH -C᎐C-), 3.90
m, CH -C᎐C), 3.79–3.58 (5H, m, H-3, H-4, H-5, H-6a, H-6b),
᎐
᎐
2
2
(1H, dd, J 6.41, 13.68, CH -C᎐C-), 3.68–3.60 (2H, m, H-2,
H-4), 3.51–3.43 (2H, m, H-6a, H-6b), 3.38–3.34 (1H, m, H-5),
2.70 (1H, br s, 4-OH), 1.91 (3H, s, COCH3); FAB-HRMS, m/z
᎐
2
found MϩHϩ 442.2226, C25H32NO6 requires 442.2230.
O r g . B i o m o l . C h e m . , 2 0 0 4 , 2, 1 6 1 7 – 1 6 2 3
1621