ppm) 7.41–7.38 (m, 10H, aromatic), 6.11 (d, 1H, 3J2,NH = 8.8 Hz,
3J2,3 = 11.2 Hz, H3), 4.04 (td, 1H, 3J2,3 = 11.2 Hz, 3J1,2 = 2.0 Hz,
H2), 3.97 (dd, 1H, 3J4,5 = 9.4 Hz, 3J5,6 = 4.0 Hz, H5), 3.86 (bd, 1H,
2J6a,6b = 11.8 Hz, 3J5,6b ª 0 Hz, H6b), 3.73 (dd, 1H, 2J6a,6b = 11.8 Hz,
3J5,6a = 4.0 Hz, H6a), 3.50 (dd, 1H, 3J4,5 = 9.4 Hz, 3J3,4 = 10.4 Hz,
H4), 3.19 (q, 12H, 3JCH2,CH3 = 7.2 Hz, 2 ¥ (CH3CH2)3NH+), 1.97
2–NH), 5.86 (d, 1H, 3J3,NH = 8.8 Hz, 3–NH), 5.69 (dd, 1H, 3J1,2
=
3
3.2 Hz, J1,P = 5.6 Hz, H1), 5.16–5.04 (m, 4H, 2 ¥ CH2Ph), 4.88
(dd, 1H, 3J4,5 = 10.4 Hz, 3J3,4 = 9.6 Hz, H4), 4.39 (ddd, 1H, 3J3,4
= 9.6 Hz, 3J2,3 = 10.0 Hz, 3J3,NH = 8.8 Hz, H3), 4.24–4.15 (m, 2H,
H2, H6a), 4.08–4.02 (m, 1H, H5), 4.91–3.87 (m, 1H, H6b), 2.07,
2.02 (s, 6H, 2 ¥ CH3CONH), 1.92, 1.75 (s, 6H, 2 ¥ CH3(CO)O);
dC (100 MHz; CDCl3; referenced to solvent peak at 77.36 ppm)
171.2, 171.0, 170.7 (s, 4C, 2 ¥ CH3CONH, 2 ¥ CH3(CO)O), 135.5
3
(s, 6H, 2 ¥ CH3CONH), 1.33 (t, 18H, JCH3,CH2 = 7.2 Hz, 2 ¥
(CH3CH2)3NH+); dC (100 MHz; CH3OH-d4; referenced to solvent
peak at 49.86 ppm) 175.2, 174.5 (s, 2C, 2 ¥ CH3CONH), 95.7 (dd,
1C, 3JC1,P = 6.1 Hz, C1), 75.8 (d, 1C, C5), 70.4 (d, 1C, C4), 63.4 (t,
1C, C6), 54.9 (d, 1C, C3), 54.8 (dd, 1C, 3JC2,P = 10.0 Hz, C2), 48.3
(t, 6C, 2 ¥ (CH3CH2)3NH+), 23.7, 23.5 (q, 2C, 2 ¥ CH3CONH),
10.0 (q, 6C, 2 ¥ (CH3CH2)3NH+); m/z (ESI+) 545 ([M + H]+,
12.2%), 444 ([M-Et3NH + 2H]+, 63.5), 267 (100); HR-MS calcd
for C22H50N4O9P [M + H]+ 545.3310, found 545.3313.
3
(ds, 2C, JC arom.,P = 6.0 Hz, aromatic), 129.2, 129.1, 129.0, 128.3,
128.3 (d, 10C, aromatic), 96.0 (dd, 1C, 2JC1,P = 6.0 Hz, C1), 70.2
2
(dt, 2C, JCH2,P = 5.4 Hz, 2 ¥ CH2Ph), 70.2 (d, 1C, C5), 67.8 (d,
1C, C4), 61.5 (t, 1C, C6), 52.9 (dd, 1C, 3JC2,P = 7.6 Hz, C2), 50.7
(d, 1C, C3), 23.4, 23.0 (q, 2C, 2 ¥ CH3(CO)O), 20.9, 20.8 (q, 2C,
2 ¥ CH3CONH); m/z (ESI+) 629 ([M + Na]+, 43.2%), 607 ([M +
H]+, 6.8), 329 (91.9), 150 (100); HR-MS calcd for C28H36N2O11P
[M + H]+ 607.2051, found 607.2053.
UDP-2,3-Diacetamido-2,3-dideoxy-a-D-glucose, diammonium
salt (2). 2,3-Diacetamido-2,3-dideoxy-a-D-glucopyranose-1-
phosphate, bistriethylammonium salt (17) (243 mg, 0.45 mmol)
and uridine 5¢-O-monophosphomorpholidate 4-morpholine-
N,N¢-dicyclohexylcarboxamidine salt (287 mg, 0.417 mmol) were
separately dried by co-evaporation with absolute pyridine (2 ¥
5 mL). Both components were dissolved in absolute pyridine
(7 mL each) and the solutions were combined. The resulting
mixture was kept without stirring at 4 ◦C for 14 days. The
conversion reached 82% (by SAX HPLC on Poros HQ50, for
details see General procedures in the Experimental section). The
mixture was concentrated to dryness under reduced pressure.
The resulting syrup was dissolved in water (1 mL) and purified
using SAX HPLC on Poros HQ20 as indicated in the General
procedures in the Experimental section to give pure diammonium
salt of 2 (138 mg, 49%). dH (400 MHz; D2O; referenced to the
methyl resonance of internal acetone at 2.22 ppm) 7.93 (d, 1H,
2,3-Diacetamido-4,6-di-O-acetyl-2,3-dideoxy-a-D-glucopyra-
nose-1-phosphate, bistriethylammonium salt (16). 2,3-Diacet-
amido-4,6-di-O-acetyl-2,3-dideoxy-a-D-glucopyranose 1-diben-
zylphosphate (15) (308 mg, 0.51 mmol) was dissolved in MeOH
(10 mL) and palladium, 10% on carbon (20 mg) was added. The
mixture was hydrogenated for 24 h at 1 atm pressure. The catalyst
was removed by filtration through 2 filter papers and washed
with MeOH. To the filtrate triethylamine (1.2 mL) was added
and the volatiles were evaporated under reduced pressure to give
pure 16 (281 mg, 88%) as a white amorphous solid. Rf = 0.59
(dichloromethane–methanol–H2O 6 : 3 : 1); [a]2D3 +53.2 (c 1.0,
MeOH); dH (400 MHz; CH3OH-d4; referenced to solvent residual
3
3
peak at 3.34 ppm) 5.52 (dd, 1H, J1,2 = 2.8 Hz, J1,P = 7.2 Hz,
H1), 5.03 (dd, 1H, J4,5 = 10.0 Hz, J3,4 = 10.0 Hz, H4), 4.43
(dd, 1H, J3,4 = 10.0 Hz, J2,3 = 10.0 Hz, H3), 4.36 (dt, 1H, J4,5
3
3
3
3
3
3
3J5,6 = 8.2 Hz, H6), 6.00 (d, 1H, J1¢,2¢ = 4.0 Hz, H1¢), 5.94 (d,
3
2
3
3
3
= 10.0 Hz, J5,6 = 2.6 Hz, H5), 4.29 (dd, 1H, J6a,6b = 9.2 Hz,
3J5,6 = 2.6 Hz, H6a), 4.17–4.13 (m, 2H, H2, H6b), 3.13 (q, 12H,
3JCH2,CH3 = 7.2 Hz, 2 ¥ (CH3CH2)3NH+), 2.06, 2.03 (s, 6H, 2 ¥
CH3CONH), 1.97, 1.88 (s, 6H, 2 ¥ CH3(CO)O), 1.32 (t, 18H,
3JCH3,CH2 = 7.2 Hz, 2 ¥ (CH3CH2)3NH+); dC (100 MHz; CH3OH-
d4; referenced to solvent peak at 49.86 ppm) 174.6, 174.4, 173.2,
172.5 (s, 4C, 2 ¥ CH3CONH, 2 ¥ CH3(CO)O), 95.2 (dd, 1C, 3JC1,P
= 5.3 Hz, C1), 71.1 (d, 1C, C4), 70.5 (d, 1C, C5), 64.2 (t, 1C,
1H, J5,6 = 8.2 Hz, H5), 5.53 (dd, 1H, J1¢¢,Pb = 7.2 Hz, J1¢¢,2¢¢ =
3.2 Hz, H1¢¢), 4.38–4.34 (m, 2H, H2¢, H3¢), 4.29–4.27 (m, 1H,
H4¢), 4.25–4.23 (m, 1H, H5¢a), 4.22–4.19 (m, 1H, H5¢b), 4.12 (dd,
1H, 3J3¢¢,4¢¢ = 9.6 Hz, 3J2¢¢,3¢¢ = 11.2 Hz, H3¢¢), 4.08 (dt, 1H, 3J2¢¢,3¢¢
=
3
4
11.2 Hz, J1¢¢,2¢¢ = 3.2 Hz, J2¢¢,Pb = 2.8 Hz, H2¢¢), 3.98 (ddd, 1H,
3J4¢¢,5¢¢ = 10.0 Hz, J5¢¢,6¢¢a = 4.0 Hz, J5¢¢,6¢¢b = 2.4 Hz, H5¢¢), 3.87
3
3
(dd, 1H, 2J6¢¢a, 6¢¢b = 12.6 Hz, 3J5¢¢,6¢¢b = 2.4 Hz, H6¢¢b), 3.81 (dd, 1H,
2J6¢¢a, 6¢¢b = 12.6 Hz, 3J5¢¢,6¢¢a = 4.0 Hz, H6¢¢a), 3.62 (dd, 1H, 3J3¢¢,4¢¢
=
3
C6), 54.6 (dd, 1C, JC2,P = 6.8 Hz, C2), 53.2 (d, 1C, C3), 47.8 (t,
9.6 Hz, 3J4¢¢,5¢¢ = 10.0 Hz, H4¢¢), 2.01, 1.98 (s, 6H, 2 ¥ CH3CONH);
dC (100 MHz; D2O; referenced to the methyl resonance of internal
acetone at 30.89 ppm) 175.4, 175.1 (s, 2C, 2 ¥ CH3CONH), 168.1
(s, 1C, C4), 153.4 (s, 1C, C2), 142.1 (d, 1C, C6), 103.3 (d, 1C,
6C, 2 ¥ (CH3CH2)3NH+), 23.6, 23.5 (q, 2C, 2 ¥ CH3(CO)O), 21.6
(q, 2C, 2 ¥ CH3CONH), 10.1 (q, 6C, 2 ¥ (CH3CH2)3NH+); m/z
(ESI+) 629 ([M + H]+, 10.1%), 528 ([M-Et3NH + 2H]+, 19.8), 102
(100); HR-MS calcd for C26H54N4O11P [M + H]+ 629.3521, found
629.3519.
2
C5), 94.6 (dd, 1C, JC1¢¢,Pb = 6.0 Hz, C1¢¢), 89.1 (d, 1C, C1¢), 83.8
3
(dd, 1C, JC4¢,Pa = 9.9 Hz, C4¢), 74.4 (d, 1C, C2¢), 73.8 (d, 1C,
2
C5¢¢), 70.3 (d, 1C, C3¢), 67.9 (d, 1C, C4¢¢), 65.7 (dt, 1C, JC5¢,Pa
=
2,3-Diacetamido-2,3-dideoxy-a-D-glucopyranose-1-phosphate,
bistriethylammonium salt (17). 2,3-Diacetamido-4,6-di-O-ace-
tyl-2,3-dideoxy-a-D-glucopyranose-1-phosphate, bistriethylam-
monium salt (16) (281 mg, 0.45 mmol) was dissolved in a 7 :
3 : 1 mixture of MeOH–H2O–Et3N (11 mL) and stirred at room
temperature. After 4 days the conversion was complete (by TLC).
The volatiles were evaporated under reduced pressure to give 17
(243 mg, quantitative) as a pale yellow solid that was sufficiently
pure for analysis. Rf = 0.41 (dichloromethane–methanol–H2O 6 :
3 : 1); [a]2D3 +27.9 (c 1.0, MeOH); dH (400 MHz; CH3OH-d4;
referenced to solvent residual peak at 3.34 ppm) 5.50 (dd, 1H,
3J1,2 = 2.0 Hz, 3J1,P = 6.0 Hz, H1), 4.23 (dd, 1H, 3J3,4 = 10.4 Hz,
5.3 Hz, C5¢), 60.9 (t, 1C, C6¢¢), 53.1 (d, 1C, C3¢¢), 52.6 (dd, 1C,
3JC2¢¢,Pb = 9.2 Hz, C2¢¢), 22.7, 22.5 (q, 2C, 2 ¥ CH3CONH); dP
(D2O, 121 MHz, referenced to external 85% phosphoric acid at 0
ppm) -7.60 (d, Pb, JPa,Pb = 20.5 Hz), -9.38 (d, Pa, JPa,Pb = 20.5
Hz); m/z (ESI+) for acid 649 ([M + H]+, 1.0%), 294 (95.9), 245
(100); m/z (ESI-) for acid 647 ([M - H]-, 20.3%), 158 (40.5), 79
(100); HR-MS calcd C19H31N4O17P2 [M + H]+ 649.1154, found
649.1151.
UDP-2,3-Diacetamido-2,3-dideoxy-a-D-glucuronic acid, tri-
ammonium and trisodium salts (1). Adam’s catalyst (PtO2; 10 mg)
was hydrogenated in water (5 mL) under atmospheric pressure
1208 | Org. Biomol. Chem., 2009, 7, 1203–1210
This journal is
The Royal Society of Chemistry 2009
©