D-Fucosamine phosphate derivatives
Russ.Chem.Bull., Int.Ed., Vol. 50, No. 7, July, 2001
1307
(2-Azido-3,4-di-O-acetyl-2-deoxy-β-D-fucopyranosyl) di-
benzyl phosphate (5). Cesium dibenzyl phosphate (670 mg,
1.57 mmol) was added with stirring at 20 °C to a solution of
compound 3 (350 mg, 1.10 mmol) in 0.7 mL of anhydrous
DMF. The reaction mixture was stirred for 2 h, poured into
5 mL of brine, and extracted with CHCl3 (3½5 mL). The
organic phase was washed with water, dried with Na2SO4, and
concentrated in vacuo, and the residue was chromatographed in
the light petroleumEtOAc system (4 : 1) to give 460 mg
(79%) of compound 5, syrup, Rf 0.27 (light petroleumEtOAc,
2 : 1). 1H NMR (CDCl3), δ: 1.18 (d, 3 H, H(6), J5,6 = 6.9);
2.03, 2.15 (both s, 3 H each, OÀñ); 3.79 (dd, 1 H, H(2),
J1,2 = 8.6 Hz, J2,3 = 10.4 Hz); 3.82 (q, 1 H, H(5)); 4.82 (dd,
1 H, H(3), J3,4 = 4.1 Hz); 5.09 (m, 5 H, H(1), PhCH2); 5.18
(br.d, 1 H, H(4)); 7.31 (m, 10 H, Ph). 13C NMR (CDCl3),
δ: 15.6 (C(6)); 20.5 (CH3CO); 60.8 (d, C(2), JC,Ð = 9.6 Hz);
was neutralized with Dowex-50 (H+) cation exchange resin to
ðH 7.5 according to ðH-meter reading. The resin was filtered
off and washed with water (3½2 mL). The filtrate was concen-
trated in vacuo to 0.1 mL and applied onto a column with TSK
HW-40, and the column was washed with water. The fractions
containing the product were combined, concentrated in vacuo
to 5 mL, and treated with Dowex-50 (Na+) cation exchange
resin. The resin was filtered off and washed with water (3½5 mL).
Lyophilization from water gave 36 mg (86%) of compound 7,
[α]D +72.1 (c 1.5, H2O). 1H NMR (D2O), δ: 1.10 (d, 3 H,
27
H(6), J5,6 = 6.6 Hz); 1.82 (s, 3 H, NHÀñ); 3.523.97 (m, 3 H,
H(2), H(3), H(4)); 4.05 (br.q, 1 H, H(5)); 5.16 (dd, 1 H, H(1),
J1,2 = 3.4 Hz, J1,P = 6.6 Hz).
Diammonium uridine 5´-(2-amino-3,4-di-O-acetyl-2-deoxy-
α-D-fucopyranosyl diphosphate) (10). Tri(n-butyl)amine
(0.073 mL, 0.308 mmol) was added with stirring at 20 °C to a
solution of uridine 5´-phosphate (100 mg, 0.308 mmol) in
anhydrous dioxane and the mixture was stired for 15 min. The
reaction mixture was lyophilized and the residue was dissolved
in 0.5 mL of anhydrous DMF. N,N´-Carbonyldiimidazole
(150 mg, 0.92 mmol) was added to the resulting solution, the
mixture was stirred under Ar for 3 h at 20 °C, and anhydrous
MeOH (0.035 mL) was added. The mixture was stirred for an
additional 20 min and evacuated (1 Torr, 20 °C) for 15 min to
remove excess MeOH. A solution of compound 7 (100 mg,
0.339 mmol) in 0.3 mL of anhydrous DMF was added at 20 °C
to the resulting solution of uridine 5´-phosphoimidazolide. The
reaction mixture was stirred for 24 h under Ar, diluted with a
25 mM aqueous NH4OAc (3 mL), and washed with CHCl3
(1½2 mL). The aqueous layer was concentrated in vacuo to
0.5 mL and applied onto a column with Sephadex G-25. The
column was washed with a 25 mM aqueous solution of NH4OAc,
the separation being monitored using an UV detector (254 nm).
The fractions containing the product were concentrated in
vacuo to 1.5 mL, and the solution was applied onto a column
with TSK HW-40, and the column was washed with water.
Lyophilization of fractions containing the product gave 91 mg
(47%) of compound 10, Rf 0.66 (propan-2-ol1 Ì aqueous
solution of NH4OAc, 2 : 1). 1H NMR (D2O), δ: 1.20 (d, 3 H,
H(6″), J5″,6″ = 6.6 Hz); 2.05, 2.20 (both s, 3 H each, OÀñ);
4.074.36 (m, 7 H, H(2″), H(2´), H(3´), H(4´), H(5´a),
H(5´b), H(5″); 5.08 (dd, 1 H, H(3″), J2″,3″ = 11.1 Hz,
68.7 (C(4)); 69.0 (d, PhCH2, JC,Ð
PhCH2, JC,Ð 4.3 Hz); 70.1 (C(3)); 71.4 (C(5)); 97.5
(d, C(1), JC,Ð 5.5 Hz); 127.7, 127.8, 128.5, 135.2,
= 4.1 Hz); 69.1 (d,
=
=
135.3, 135.5 (Ar); 169.6, 170.2 (COCH3). 31Ð NMR (CDCl3):
δ 2.75.
(2-Azido-3,4-di-O-acetyl-2-deoxy-α-D-fucopyranosyl) di-
benzyl phosphate (6). BF3Et2O (32 µL, 0.25 mmol) was
added with stirring at 20 °C to a solution of compound 5
(248 mg, 0.49 mmol) and 2-bromopyridine (151 mg, 0.98 mmol)
in 2 mL of anhydrous CH2Cl2. The mixture was stirred for 1 h,
the solvent was evaporated in vacuo, and the residue was
chromatographed in the light petroleumEtOAc system (5 : 1)
to give 178 mg (72%) of compound 6, Rf 0.36 (heptaneEtOAc,
25
1
:
1), m.p. 9495 °C, [α]D
+77.5 (c 2.35, CHCl3).
Found (%): C, 52.01; H, 5.49; N, 8.26. C22H28N3O9Ð. Calcu-
lated (%): C, 51.87; H, 5.54; N, 8.23. 1H NMR (C6D6), δ: 0.77
(d, 3 H, H(6), J5,6 = 6.8 Hz); 1.65, 1.73 (both s, 3 H each,
OÀñ); 3.60 (ddd, 1 H, H(2), J1,2 = 3.4 Hz, J2,3 = 11.1 Hz,
J1,P = 3.1 Hz); 3.78 (br.q, 1 H, H(5)); 4.95, 5.12 (both d, 2 H
each, CH2Ðh, JH,Ð = 8.1 Hz); 5.22 (br.d, 1 H, H(4)); 5.45 (dd,
1 H, H(3), J3,4 = 4.1 Hz); 5.95 (dd, 1 H, H(1), J1,P = 6.3 Hz);
7.007.35 (m, 10 H, Ph). 13C NMR (CDCl3), δ: 15.6 (C(6));
20.5, 20.6 (both CH3CO); 57.3 (d, C(2), JC,Ð = 8.6 Hz); 66.8
(C(3)); 68.7 (C(4)); 69.4 (C(5)); 69.5, 69.6 (both d, PhCH2,
JH,Ð = 5.6 Hz); 96.1 (d, C(1), JC,Ð = 5.6 Hz); 127.8, 127.9,
128.6, 135.4, 135.5 (Ar); 169.7, 170.2 (both COCH3). 31Ð NMR
(CDCl3): δ 2.05.
J
3″,4″ = 3.4 Hz); 5.23 (br.d, 1 H, H(4″)); 5.50 (dd, 1 H, H(1″),
2-Amino-3,4-di-O-acetyl-2-deoxy-α-D-fucopyranosyl phos-
phate (7). Compound 6 (178 mg, 0.354 mmol) was dissolved in
3 mL of anhydrous dioxane and hydrogenated at 25 °C under
atmospheric pressure over 10% Pd(OH)2/C (40 mg) for 4 h.
The catalyst was filtered off and washed with dioxane (1½5 mL)
and EtOH (2½5 mL). Lyophilization of the solution gave 91 mg
(87%) of compound 7, Rf 0.32 (propan-2-ol1 Ì aqueous
NH4OAc, 2 : 1). 1H NMR (D2O), δ: 0.94 (d, 3 H, H(6),
J5,6 = 6.6 Hz); 1.82, 1.96 (both s, 3 H each, OÀñ); 3.62 (br.dd,
1 H, H(2), J1,2 = 3.4 Hz, J2,3 = 10.8 Hz); 4.31 (br.q, 1 H,
H(5)); 5.105.21 (m, 2 H, H(3), H(4)); 5.51 (dd, 1 H, H(1),
J1,P = 6.6 Hz). 13C NMR (D2O), δ: 16.2 (C(6)); 21.2, 21.3
(both CH3CO); 49.8 (d, C(2), JC,Ð = 9.2 Hz); 67.5 (C(3)); 69.5
(C(4)); 71.2 (C(5)); 93.3 (d, C(1), JC,Ð = 5.1 Hz); 173.9, 174.9
(both COCH3). 31Ð NMR (CDCl3): δ 0.56.
Disodium 2-acetamido-2-deoxy-α-D-fucopyranosyl phos-
phate (9). A solution of N-acetoxysuccinimide (NAS)17 (44 mg,
0.29 mmol) in 0.44 mL of aqueous THF (1 : 1) was added with
stirring at 20 °C to a solution of compound 7 (42 mg, 0.14 mmol)
in 0.22 mL of water. The reaction mixture was stirred for 2 h,
pH 7.5 beng maintained by addition of 0.1 Ì NaOH, and
cooled to 0 °C. A 1 Ì solution of NaOH (0.1 mL) was added,
and the mixture was kept for 1 h at 20 °C. The reaction mixture
J1″,2″ = 3.4 Hz, J1″,P = 6.6 Hz); 5.78 (m, 2 H, H(5), H(1´));
7.73 (d, 1 H, H(6), J5,6 = 8.3 Hz). 13C NMR (D2O),
δ: 16.2 (C(6″)); 21.1, 21.3 (both COCH3); 49.7 (d, C(2″),
JC,Ð = 9.3 Hz); 65.3 (d, C(5´), JC,Ð = 5.4 Hz); 67.8 (C(3″);
69.5 (C(4″)); 70.7 (C(5″)); 71.1 (C(2´)); 75.1 (C(3´)); 84.1 (d,
C(4´), JC,Ð
= 8.8 Hz); 89.7 (C(1´)); 94.0 (d, C(1″),
JC,Ð = 5.4 Hz); 103.7 (C(5)); 142.9 (C(6)); 152.9 (C(2)); 167.3
(C(4)); 173.8, 174.8 (both COCH3). 31Ð NMR (D2O), δ: 10.1,
12.9 (both d, J = 19.3 Hz).
Disodium uridine 5´-(2-acetamido-2-deoxy-α-D-fuco-
pyranosyl diphosphate) (1). A solution of NAS (22.4 mg,
0.12 mmol) in 0.4 mL of aqueous THF (1 : 1) was added with
stirring at 20 °C to a solution of compound 10 (24 mg,
0.038 mmol) in 0.3 mL of water. The reaction mixture was
stirred for 5 h, pH 7.5 being maintained by addition of 0.1 Ì
NaOH, and cooled to 0 °C. A 1 Ì solution of NaOH (0.2 mL)
was added, and the mixture was kept for 3 h at 20 °C and
neutralized with Dowex-50 (H+) cation exchange resin to
ðH 7.5 according to ðH-meter reading. The cation exchange
resin was filtered off and washed with water (3½3 mL), and the
filtrate was concentrated in vacuo to 0.2 mL and subjected to
gel chromatography on a column with TSK HW-40; water was
used for elution. The fractions containing the product were