Inhibitors of â1,4-Galactosyltransferase
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 19 6063
3.86 (t, 1 H, J5,6 ) 6.2 Hz, H-5), 3.62-3.70 (m, 8 H, [OCH2-
CH2]), 3.48-3.61 (m, 6 H, H-6a, H-6b, [OCH2CH2]), 2.10 (s, 6
H, Ac × 2), 2.00 (s, 3 H, Ac); 13C NMR (150 MHz, CDCl3, 27
°C) δ 170.12, 169.99, 169.50, 135.82, 133.29, 132.99, 132.08,
128.90, 128.14, 127.88, 127.69, 126.47, 126.07, 125.81, 86.62,
76.02, 73.33, 72.21, 71.13, 70.69, 70.66, 70.52, 69.48, 69.20,
67.81, 67.56, 20.85, 20.67, 20.61; HRMS calcd for [M + H]+
C35H43O11S 671.2526, found 671.2548.
CHCH2-, H-2), 4.72 (s, 2 H, OCH2-naphthalene), 4.55-4.60
(m, 4 H, CH2dCHCH2- × 2), 4.35-4.37 (t, 1 H, J5,6a ) 6.3
Hz, H-5), 3.56-3.69 (m, 13 H, [OCH2CH2], H-6b), 3.46-49 (dd,
1 H, J6a,b ) 10.2 Hz, H-6a), 2.12 (s, 3 H, Ac), 2.08 (s, 3 H, Ac),
1.99 (s, 3 H, Ac); 13C NMR (150 MHz, CDCl3, 27 °C) δ 170.03,
169.94, 169.87, 135.82, 133.28, 132.98, 132.24, 132.19, 132.13,
128.11, 127.85, 127.67, 126.43, 126.05, 125.80, 125.79, 118.66,
118.53, 94.56, 73.30, 71.02, 70.66, 70.44, 69.57, 69.47, 68.93,
68.54, 68.51, 68.45, 68.41, 68.02, 67.27, 67.22, 67.17, 20.69,
20.63, 20.58; HRMS calcd for [M + H]+ C35H48O15P 739.2731,
found 739.2701.
2,3,4-Tri-O-acetyl-6-O-(10-[2-naphthyl]-3,6,9-trioxade-
canyl)-D-galactopyranose (24). To a solution of 23 (500 mg,
745 µmol) in mixed solvent (acetone/H2O ) 20:1, 10 mL) was
added N-bromosuccinimide (398 mg, 2.23 mmol) at room
temperature. After being stirred for 90 min, the mixture was
concentrated and dissolved in CHCl3. The solution was washed
with saturated NaHCO3 and brine, and the organic layer was
dried by MgSO4, filtered, and concentrated. The residue
obtained was purified by silica gel column chromatography
(hexane/EtOAc ) 3:1 to 1:2) to give 24 (333 mg, 77%) as a
syrup. 1H NMR (600 MHz, CDCl3, 27 °C) δ 7.83 (m, 4 H,
naphthalene), 7.45-7.48 (m, 3 H, naphthalene), 5.49 (m, 0.8
H, H-3R), 5.41-5.43 (m, 1 H, H-1R, H-4â), 5.16 (dd, 0.8 H,
H-2R), 5.09 (dd, 0.2 H, H-2â), 5.05 (dd, 0.2 H, H-3â), 4.74 (s, 2
H, OCH2-naphthalene), 4.67 (t, 0.2 H, H-1â), 4.48 (dd, 0.8 H,
H-5R), 4.24 (s, 0.8 H, H-4R), 3.86 (t, 0.2 H, H-5â), 3.57-3.72
(m, 12 H, [OCH2CH2] × 3), 3.43-3.55 (m, 2 H, H-6a, H-6b),
1.92-2.13 (m, 9 H, Ac × 3); 13C NMR (150 MHz, CDCl3, 27
°C) δ 170.75, 170.70, 170.53, 170.45, 170.25, 169.84, 135.95,
135.90, 133.51, 133.23, 128.40, 128.11, 127.93, 126.74, 126.32,
126.10, 126.04, 95.88, 90.73, 73.52, 72.39, 71.26, 71.21, 71.15,
71.05, 70.90, 70.87, 70.82, 70.73, 70.67, 70.62, 70.14, 70.05,
69.62, 69.34, 68.89, 67.94, 67.32, 21.05, 20.92, 20.89, 20.83;
HRMS calcd for [M + Na]+ C29H38O12Na 601.2261, found
601.2237.
2,3,4-Tri-O-acetyl-6-O-(10-[2-naphthyl]-3,6,9-trioxade-
canyl)-r-D-galactopyranosyl Phosphate Mono-Et3N Salt
(27). To a solution of 26 (60 mg, 83 µmol) in THF (2 mL) was
added Et2NH (172 µL, 1.66 mmol) and tetrakis(triphenylphos-
phine)palladium (19 mg, 16.6 µmol) at room temperature.37
After being stirred for 2 h at room temperature, the solution
was concentrated and the residue obtained was purified by
Iatrobeads column chromatography (CHCl3/MeOH ) 2:1,
containing 0.5 vol % Et3N) to give 27 (57 mg, 91%). [R]20
D
1
+40.9° (c 1.00, CHCl3). H NMR (500 MHz, CDCl3, 27 °C) δ
7.82-7.86 (m, 4 H, naphthalene), 7.31-7.52 (m, 3 H, naph-
thalene), 5.79 (dd, 1 H, J1,P ) 7.7 Hz, J1,2 ) 3.3 Hz, H-1), 5.53
(d, 1 H, H-4), 5.44 (dd, 1 H, J3,4 ) 3.3 Hz, H-3), 5.16 (dt, 1 H,
J2,3 ) 10.7 Hz, H-2), 4.76 (s, 2 H, OCH2-naphthalene), 4.49 (t,
1 H, J5,6a ) 6.6 Hz, H-5), 3.53-3.73 (m, 13 H, [OCH2CH2],
H-6a), 3.48 (dd, 1 H, J5,6b ) 7.8 Hz, J6a,b ) 9.5 Hz, H-6b), 3.03
(dd, 6 H, CH3CH2N × 3), 2.15 (s, 3 H, Ac), 2.10 (s, 3 H, Ac),
1.99 (s, 3 H, Ac), 1.30 (t, 9 H, CH3CH2N × 3); 13C NMR (150
MHz, CDCl3, 27 °C) δ 170.79, 170.20, 169.96, 135.86, 133.29,
132.99, 128.13, 127.87, 127.68, 126.45, 126.07, 125.83, 92.04,
91.99, 73.30, 70.77, 70.62, 70.59, 70.54, 70.34, 69.50, 68.88,
68.65, 68.20, 68.16, 67.81, 67.50, 45.44, 41.44, 20.95, 20.72,
11.17, 8.95; HRMS calcd for [M + Na]+ C29H39O15PNa 681.1924,
found 681.1937.
Diallyl 2,3,4-Tri-O-acetyl-6-O-(10-[2-naphthyl]-3,6,9-tri-
oxadecanyl)-r-D-galactopyranosyl Phosphite (25). To a
solution of 24 (106 mg, 183 µmol) in CH3CN (2 mL) was added
1H-tetrazole (23 mg, 329 µmol), and the mixture was cooled
to -40 °C followed by the addition of N,N-diallyl-diisopropyl
phosphoramidite (73 µL, 274 µmol).36 After the mixture was
stirred for 7 h at -40 °C, EtOAc was added and the mixture
was washed with saturated NaHCO3 and brine. The organic
layer was dried by MgSO4, filtered, and concentrated. The
residue obtained was purified by silica gel column chroma-
tography (hexane/EtOAc ) 4:1 to 3:1, containing 0.5 vol %
6-O-(10-[2-Naphthyl]-3,6,9-trioxadecanyl)-r-D-galacto-
pyranosyl Phosphate Mono-Et3N Salt (28). To a solution
of 27 (57 mg, 75 µmol) in MeOH (2 mL) was added Et2NH
(500 µL), and the mixture was stirred for 3 days at room
temperature. The reaction mixture was concentrated, and the
residue obtained was purified by ion exchange column chro-
matography (DEAE Sephasell, 10-50 mM NH4HCO3 as a
eluent). The obtained residue redissolved in H2O was passed
through a cation-exchange resin column (Dowex 50W-X8, H+
Et3N) to give 25 (65 mg, 49%). [R]20 +17.0° (c 1.00, CHCl3).
form followed by Et3NH+ form) to give 28 (44 mg, 88%). [R]20
D
D
1H NMR (500 MHz, CDCl3, 27 °C) δ 7.78-7.83 (m, 4 H,
naphthalene), 7.26-7.48 (m, 3 H, naphthalene), 5.88-5.96 (m,
+41.6° (c 1.00, D2O). 1H NMR (500 MHz, D2O, 27 °C) δ 7.87-
7.95 (m, 4 H, naphthalene), 7.52-7.59 (m, 3H, naphthalene),
5.52 (dd, 1H, J1,P ) 7.3 Hz, J1,2 ) 3.6 Hz, H-1), 4.72 (s, 2 H,
OCH2-naphthalene), 4.20 (t, 1 H, J5,6 ) 6.0 Hz, H-5), 3.93 (d,
1 H, H-4), 3.88 (dd, 1 H, J3,4 ) 3.3 Hz, H-3), 3.80 (dt, 1 H, J2,3
) 10.3 Hz, H-2), 3.61-3.71 (m, 14 H, H-6a, H-6b, [OCH2CH2]),
3.18 (dd, 8 H, CH3CH2N × 4), 1.27 (t, 12 H, CH3CH2N × 4);
13C NMR (125 MHz, D2O, 27 °C) δ 137.70, 135.65, 135.52,
131.05, 130.65, 130.43, 129.97, 129.33, 129.24, 129.14, 97.67,
97.62, 75.52, 72.60, 72.34, 72.27, 72.20, 72.16, 72.00, 71.73,
71.42, 71.06, 71.00, 49.36, 10.93; HRMS calcd for [M + H +
Et3N]+ C29H49NO12P 634.2992, found 634.3003.
2 H, CH2dCHCH2- × 2), 5.74 (dd, 1 H, J1,P ) 8.4 Hz, J1,2
)
3.4 Hz, H-1), 5.50 (d, 1 H, H-4), 5.36 (dd, 1 H, J2,3 ) 10.8 Hz,
J3,4 ) 3.3 Hz, H-3), 5.17-5.33 (m, 5 H, CH2dCHCH2- × 2,
H-2), 4.73 (s, 2 H, OCH2-naphthalene), 4.34-4.41 (m, 5 H,
CH2dCHCH2- × 2, H-5), 3.54-3.71 (m, 12 H, [OCH2CH2]),
3.44-3.54 (m, 2 H, J6a,b ) 11.0 Hz, H-6a, H-6b), 2.12 (s, 3 H,
Ac), 2.04 (s, 3 H, Ac), 1.98 (s, 3 H, Ac); 13C NMR (125 MHz,
CDCl3, 27 °C) δ 170.25, 170.12, 170.00, 135.81, 134.37, 134.35,
133.29, 132.99, 128.14, 127.87, 127.69, 126.47, 126.06, 125.82,
116.82, 116.75, 90.93, 90.80, 73.34, 71.01, 70.68, 70.64, 70.46,
69.47, 69.19, 68.48, 68.26, 67.61,63.47, 63.40, 63.32, 63.26,
20.77, 20.71, 20.66; HRMS calcd for [M]+ C35H47O14P 722.2703,
found 722.2692.
Uridine 5′-(6-O-[10-{2-Naphthyl}-3,6,9-trioxadecanyl]-
r-D-galactopyranosyl) Diphosphate Disodium Salt (2).
Compound 28 (26 mg, 39 µmol) was dissolved in dry pyridine
(1 mL) and concentrated in vacuo three times to remove water.
To the residual syrup 28 in dry pyridine (1 mL) was added
1H-tetrazole (11 mg, 156 µmol) and UMP-morpholidate (54 mg,
78 µmol, 1 mL in pyridine solution) at room temperature. After
being stirred for 2 days, the mixture was evaporated and the
residue obtained was subjected to purification by ion exchange
column chromatography (DEAE-Sephasell, 10-150 mM NH4-
HCO3 as the eluent). Further purification by gel permeation
column chromatography (Sephadex G-15, water as an eluent)
and cation-exchange resin column (Dowex 50W-X8, Na+ form)
afforded compound 2 (14 mg, 41%). The compound was
lyophilized to give an amorphous powder. 1H NMR (500 MHz,
D2O, 27 °C) δ 7.87-7.97 (m, 6 H, naphthalene, H-5), 7.55-
7.61 (m, 3 H, naphthalene), 5.90 (d, 1 H, J1′,2′ ) 4.8 Hz, H′-1),
Diallyl 2,3,4-Tri-O-acetyl-6-O-(10-[2-naphthyl]-3,6,9-tri-
oxadecanyl)-r-D-galactopyranosyl Phosphate (26). To a
solution of 25 (59 mg, 82 µmol) in THF (2 mL) was added Et3N
(114 µL, 820 µmol) and cooled to -10 °C followed by the
addition of tert-butyl hydroperoxide (49 µL, 246 µmol). After
being stirred for 4 h at -10 °C, the reaction mixture was
concentrated and the residue obtained was dissolved in CHCl3.
The solution was washed with saturated NaHCO3 and brine.
The organic layer was dried by MgSO4, filtered, and concen-
trated to give 26 (60 mg, 98%). [R]20 +47.8° (c 1.00, CHCl3).
D
1H NMR (600 MHz, CDCl3, 27 °C) δ 7.78-7.82 (m, 4 H,
naphthalene), 7.45-7.48 (m, 3 H, naphthalene), 5.91-5.95 (m,
3 H, H-1, CH2dCHCH2- × 2), 5.52 (d, 1 H, H-4), 5.33-5.40
(m, 3 H, CH2dCHCH2-, H-3), 5.23-5.28 (m, 3 H, CH2d