13300 J. Am. Chem. Soc., Vol. 120, No. 51, 1998
Hanessian et al.
EtOAc-Et3N (100:50:1) as an eluant to provide the title compound
reaction mixture was warmed to room temperature within 10 min and
stirred for 3 h until 3 was completely consumed. The reaction mixture
was cooled to 0 °C, diluted with cold water (30 mL) and 0.5 M NH4-
HCO3 (5 mL), and passed through a bed of Celite. The filtrate and
washings were combined and lyophilized. The residue was dissolved
in deionized water (5 mL), then treated with alkaline phosphatase
(Boehringer Mannheim, 500 µL, 1 unit/µL), and the mixture was kept
at room temperature until the unreacted UDP was no longer detected
by TLC. The mixture was diluted with water (20 mL) and loaded onto
a Dowex-1×2-200 column (bicarbonate form, 2.5 × 12 cm). The
column was first eluted with water (400 mL) to remove the neutral
compounds, then eluted with a linear gradient of 0.05 M ammonium
bicarbonate (900 mL) and 0.5 M NH4HCO3 (200 mL). Fractions which
contained UDP-Gal were pooled and concentrated to 10 mL at 25 °C.
The concentrated solution was neutralized to pH 7 with Dowex 50W×8
resin. After removal of the resin by filtration, and lyophilization, the
desired UDP-Gal was obtained as a white powder27 (∼60%, R/â ratio
4:1). A portion was converted to the sodium salt by passage over
Dowex-50W×8 (Na+ form), elution with water, and lyophilization: 1H
NMR (D2O, 400 MHz) δ 7.94 (d, J ) 8.1 Hz, 1H, H-6”), 5.96 (d, J )
8.1 Hz, 1H, H-5”), 5.94 (d, J ) 4.2 Hz, 1H, H-1′), 5.62 (dd, J ) 7.0,
3.0 Hz, H-1 of R-Gal), 4.94 (dd, J ) 7.4, 7.2 Hz, H-1 of â-Gal), 4.20
(d, J ) 2.5 Hz, H-4), 3.62 (dd, J ) 9.6, 7.3 Hz, H-2 of â-Gal); 13C
NMR (D2O) δ 166.95, 152.54, 142.33, 103.37, 96.55 (d, J ) 6.9 Hz,
C1), 89.07, 83.91, 76.56, 74.48, 72.61, 70.36, 70.02, 69.12 (d, J ) 8.5
Hz, C2), 65.64 (d, J ) 5.8 Hz, C5′), 61.91, 61.71; 31P NMR (D2O) δ
-10.49 and -12.03; ESI MS m/z 633 (M + Na)+, 611 (M + H)+. The
ammonium salt of 13 was not suitable for MS analysis.
1
(132 mg, 90%) as a white foam: [R]D -191.6 (c 0.80, CH2Cl2); H
NMR (CDCl3, 400 MHz) δ 8.15, 7.88, 7.82 (ddd, J ) 8.0, 1.5, 1.0 Hz,
each 2H, benzoyl C2-H, C6-H), 7.75 (dd, J ) 4.9, 1.5 Hz, 1H, MOP
H-4), 7.61 (dddd, J ) 8.0, 7.8, 1.2, 1.0 Hz, 1H, Ar-H), 7.49 (dddd, J
) 8.0, 7.8, 1.2, 1.0 Hz, 2H, Ar-H), 7.48-7.41 (m, 2H, Ar-H), 7.29,
7.26 (ddd, J ) 8.0, 7.8, 0.8 Hz, each 2H, Ar-H), 7.01 (dd, J ) 7.8, 1.5
Hz, 1H, MOP H-6), 6.90 (dd, J ) 7.8, 4.9 Hz, 1H, MOP H-5), 6.53
(d, J1,2 ) 8.3 Hz, 1H, H-1), 6.12 (dd, J2,3 ) 10.4 Hz, 1H, H-2), 5.81
(dd, J3,4 3.5 Hz ) J4,5 1.0 Hz, 1H, H-4), 5.71 (dd, 1H, H-3), 4.35 (dq,
J5,6 ) 6.4 Hz, 1H, H-5), 3.70 (s, 3H, OCH3), 1.38 (d, 3H, H-6); 13C
NMR (100.6 MHz, CDCl3) δ 166.0, 165.6, 165.1 (CdO), 152.0 (MOP
C2), 144.2, 136.7, 133.3, 133.1, 132.9, 130.01, 129.99, 129.97, 129.95,
129.72, 129.70, 129.68, 129.66, 129.65, 129.60, 129.59, 129.57, 129.43,
129.18, 128.87, 128.47, 128.45, 128.43, 128.21, 128.19, 128.17, 128.16,
128.14, 128.13, 128.11, 128.10, 128.08, 119.13, 118.82 (aromatic C,
CH), 94.55 (C-1), 72.4, 71.0, 70.5, 69.4 (C2, C3, C-4, C5), 55.7 (OCH3),
16.3 (C6); FAB MS (m/z) 584 (M + H) +; exact FAB MS calcd for
C33H30NO9 (M + H)+ 584.1920, found 584.1921.
Dibenzyl 2,3,4-Tri-O-benzoyl-â-L-fucopyranosyl 1-Phosphate (11).
To a mixture of 10 (58.4 mg, 0.10 mmol), dibenzyl phosphate (55.7
mg, 0.20 mmol), crushed 4-Å molecular sieves (584 mg), and anhydrous
CH2Cl2 (10 mL) was added copper triflate (29 mg, 0.08 mmol) in five
portions within 40 min under argon. The reaction was monitored by
TLC and neutralized with pyridine. The suspension was filtered through
a Celite pad, and the filtrate was concentrated. The residue was
subjected to silica gel chromatography using CH2Cl2-acetone-Et3N
(98:2:1) as an eluant to give the title compound (38 mg, 51%);8,25 the
R-anomer (7.4 mg, 10%), and unreacted starting material (6 mg, 10%):
1H NMR (400 MHz, CDCl3) δ 8.12, 7.96, 7.80 (dd, J ) 8.0, 1.2 Hz,
each 2H, benzoyl C2-H, C6-H), 7.68-6.99 (m, 19H, Ar-H), 5.89 (dd,
Uridine 5′-Diphospho-D-glucose (UDP-Glc, 14). UDP-Glc was
prepared from UDP free acid and GlcOMOP using the same procedure
as described above for the preparation of UDP-Gal except that the time
of reaction was extended to 6 h. UDP-Glc was obtained as a white
powder (∼50%, R/â ratio 3:1): 1H NMR (D2O, 400 MHz) δ 7.92 (d,
J ) 8.0 Hz, 1H, H-6′′), 5.99 (d, 1H, J ) 3.5 Hz, 1H, H-1′), 5.94 (d, J
) 8.0 Hz, 1H, H-5′′), 5.60 (dd, J ) 7.3, 3.3 Hz, H-1 of R-Glc), 5.01
(dd, J ) 8.0, 7.9 Hz, H-1 of â-Gal), 3.78 (dd, J ) 9.8, 9.8 Hz, H-3),
3.53 (ddd, J ) 9.8, 3.3, 3.0 Hz, H-2 of R-Glc), 3.47 (dd, J ) 9.8, 9.8
Hz, H-4), 3.39 (ddd, J ) 9.8, 8.0, 3.0 Hz, H-2 of â-Gal); 13C NMR
(D2O) δ 166.94, 152.52, 142.28, 103.39, 103.33, 98.55 (C1 of â-Glc),
96.27 (d, J ) 6.7 Hz, C1 of R-Glc), 89.02, 88.92, 83.94, 83.85, 77.17,
74.44, 74.25, 73.49, 70.40, 70.30, 70.10, 69.82, 65.62, 65.56, 61.46,
60.92; 31P NMR (D2O) δ -10.62 and -12.26; ESI MS m/z 633 (M +
Na)+, 611 (M + H)+.
J2,3 ) 10.4 Hz, J1,2 ) 8.0 Hz, 1H, H-2), 5.76 (dd, J3,4 ) 3.2 Hz, J4,5
)
1.0 Hz, 1H, H-4), 5.67 (dd, J1,P ) 7.2 Hz, 1H, H-1), 5.57 (dd, 1H,
H-3), 5.14, 5.12 (ABM, J ) 11.8 Hz, 7.4 Hz, each 1H, PhCH2), 4.86,
4.77 (ABM, J ) 11.6, 6.4 Hz, each 1H, PhCH2), 4.22 (dq, J5,6 ) 6.4
Hz, 1H, H-5), 1.35 (d, 1H, H-6); 13C NMR (CDCl3) δ 165.7, 165.4,
165.2 (CdO), 135.5, 135.4, 135.04, 134.96, 133.45, 133.35, 133.21,
129.89, 129.73, 129.68, 129.03, 128.80, 128.63, 128.53, 128.39, 128.26,
128.21, 127.83, 127.35 (aromatic C, CH), 96.94 (d, J1,P ) 4.7 Hz, C-1),
71.7, 70.8, 70.5 (C3, C4, C5), 69.6 (d, J2,P ) 9.3 Hz, C-2), 69.5 (d,
JC,P ) 5.6 Hz, PhCH2), 69.2 (d, JC,P ) 5.6 Hz, PhCH2), 16.1 (C6); 31
P
NMR (CDCl3) δ -2.41; FAB MS (m/z) 737 (M + H)+; exact FAB
MS calcd for C41H38O11P (M + H)+ 737.2156, found 737.2154.
Uridine 5′-Diphospho-D-galactose (UDP-Gal, 13). To a suspension
of UDP trisodium salt dihydrate (500 mg, 1 mmol) in DMF (16 mL)
at 4 °C, was added Amberlite IR-120 (H+ form) resin until the UDP
was completely dissolved. The resin was filtered and washed with DMF.
The combined filtrate and washing were concentrated in vacuo, the
residue was dissolved in anhydrous DMF (8 mL), and the solution was
concentrated. This process was repeated three times. The residue was
then dissolved in anhydrous DMF (4 mL), and crushed 4-Å molecular
sieves (500 mg) were added to the solution. The resulting mixture was
stirred at room temperature for 2 h, cooled to 0 °C, and treated with a
solution of 3 (152 mg, 0.5 mmol) in DMF (2 mL) dropwise. The
Acknowledgment. We thank NSERCC for generous finan-
cial support through the Medicinal Chemistry Chair Program.
Supporting Information Available: 1H, 13C, and 31P NMR
spectra for all compounds (24 pages, print/PDF). See any current
masthead page for ordering information and Internet access
instructions.
JA982783I