CMP-Sialic Acid Derivatives To Probe (Poly)sialyltransferases
21.0, 20.8 (4CH3-acetyl), 11.2 ppm (C3-propanoyl); IR: n˜ =3295,
2926, 1737, 1659, 1540, 1439, 1369, 1217, 1128, 1033, 953, 753,
692, 602 cmÀ1; HRMS (FAB): m/z calcd. for C27H35NO12S: 597.1880
[M]; found: 597.1872.
fonic acid (1.00 mL, 1.48 g, 15.4 mmol) and cyclopropanecarboxylic
acid anhydride (479 mg, 3.11 mmol). Subsequent peracetylation
was achieved with acetic anhydride (2.00 mL, 2.16 g, 21.0 mmol) in
pyridine (12 mL). The reaction sequence delivered the product in
a yield of 29% (305 mg, 0.500 mmol) as a colourless syrup. [a]D20
=
Synthesis of thiophenyl(methyl-5-N-butanoyl-4,7,8,9-tetra-OAc-
1
3
+378 (c=1.0, CHCl3); H NMR (400 MHz, C6D6): d=7.61 (dd, JH,H
=
3,5-dideoxy-a-d-glycero-d-galacto-2-nonulopyranosylate)
(10):
4
8.2 Hz, JH,H =1.2 Hz, 1H; Hortho), 7.28–7.17 (m, 2H; Hmeta), 7.15–7.06
(m, 1H; Hpara), 5.65–5.60 (m, 1H; H8), 5.50 (dd, 3JH,H =7.0, 2.3 Hz,
The reaction was carried out according to the general procedure
with thioglycoside 8 (1.00 g, 1.73 mmol), methanesulfonic acid
(1.00 mL, 1.48 g, 15.4 mmol) and butyric acid anhydride (0.510 mL,
0.491 mg, 3.10 mmol). Subsequent peracetylation was achieved
with acetic anhydride (2.00 mL, 2.16 g, 21.0 mmol) in pyridine
(12 mL). The reaction sequence delivered the product in a yield of
45% (477 mg, 0.780 mmol) as a colourless syrup. [a]2D0 =À168 (c=
1H; H7), 4.92–4.89 (m, 1H; H4), 4.73 (dd, 2JH,H =12.1 Hz, JH,H
=
3
2.8 Hz, 1H; H9), 4.43 (dd, 2JH,H =12.1 Hz, 3JH,H =5.8 Hz, 1H; H9),
3
4.38–4.33 (m, 1H; H5), 3.95 (dd, JH,H =10.6, 2.3 Hz, 1H; H6), 3.23 (s,
3H; OCH3), 2.96 (dd, 2JH,H =12.6 Hz, 3JH,H =4.9 Hz, 1H; H3eq), 2.06–
2.01 (m, 1H; H3ax), 1.96, 1.93, 1.78, 1.55 (4s, 4ꢂ3H; 4CH3-acetyl),
1.54–1.48 (m, 1H; CH-cyclopropylcarbonyl), 0.93–0.89 (m, 2H; CH2-
cyclopropylcarbonyl), 0.48–0.43 ppm (m, 2H; CH2-cyclopropylcar-
bonyl); 13C NMR (101 MHz, C6D6): d=165.8 (C1), 137.6 (Cortho), 129.9
(Cmeta), 129.4 (Cpara), 90.3 (C2), 76.5 (C6), 71.0 (C8), 69.8 (C4), 68.8
(C7), 63.1 (C9), 52.2 (OCH3), 49.0 (C5), 39.8 (C3), 21.0, 20.9, 20.8,
20.6 (4CH3-acetyl), 14.3 (CH-cyclopropylcarbonyl), 10.8, 10.1 ppm
(2CH2-cyclopropylcarbonyl); IR: n˜ =3373, 2950, 1692, 1655, 1498,
1463, 1372, 1201, 1154, 1048, 751, 675, 574 cmÀ1; HRMS (FAB): m/z
calcd. for C28H35NO12S: 609.2 [M]; found: 609.4.
1
3
1.0, CHCl3); H NMR (400 MHz, C6D6): d=7.64 (d, JH,H =7.4 Hz, 2H;
Hortho), 7.21–7.14 (m, 2H; Hmeta), 7.10–7.02 (m, 1H; Hpara), 5.68–5.65
3
3
(m, 1H; H8), 5.50 (dd, JH,H =7.0, 2.0 Hz, 1H; H7), 4.88 (ddd, JH,H
=
11.2, 11.2, 4.7 Hz, 1H; H4), 4.76 (dd, 2JH,H =12.4 Hz, 3JH,H =2.6 Hz,
1H; H9), 4.47 (dd, 2JH,H =12.4 Hz, JH,H =5.8 Hz, 1H; H9), 4.38–4.30
3
(m, 1H; H5), 3.99 (dd, 3JH,H =10.8, 1.8 Hz, 1H; H6), 3.21 (s, 3H;
OCH3), 3.00 (dd, 2JH,H =12.8 Hz, 3JH,H =4.7 Hz, 1H; H3eq), 2.05–2.01
(m, 1H; H3ax), 1.96, 1.93, 1.78, 1.57 (4s, 4ꢂ3H; 4CH3-acetyl), 1.88–
1.83 (m, 1H; CH2-butanoyl), 1.71–1.66 (m, 1H; CH2-butanoyl), 1.62–
3
1.51 (m, 2H; CH2-butanoyl), 0.83 ppm (dd, 3JH,H =7.3 Hz, JH,H
=
General procedure for the cleavage of thioglycosides: The ap-
propriate N-alkanoyl neuraminic acid thioglycoside (compound 9,
10, 11 or 12, 1.0 equiv) was dissolved in acetone und distilled
water (20:1). N-Bromosuccinimide (NBS, 2.3 equiv) was added, and
the reaction mixture was stirred for 3 h at room temperature. After
removal of the solvent, residues were dissolved in chloroform. The
solution was washed with saturated sodium hydrogen carbonate
solution and distilled water. The organic fraction was dried over
sodium sulfate. Column chromatography was conducted with pe-
troleum ether and acetone.
7.3 Hz, 3H; CH3-butanoyl); 13C NMR (101 MHz, C6D6): d=172.6 (C1),
137.0 (Cortho), 130.2 (Cmeta), 129.5 (Cpara), 75.8 (C6), 71.2 (C8), 70.1
(C4), 68.3 (C7), 62.8 (C9), 52.6 (OCH3), 49.3 (C5), 39.4 (C3), 38.8 (C2-
butanoyl), 20.9, 20.6, 20.4, 20.4 (4CH3-acetyl), 19.4 (C3-butanoyl),
14.9 ppm (C4-butanoyl); IR: n˜ =3360, 2962, 1737, 1656, 1537, 1439,
1369, 1218, 1129, 1034, 753, 692, 603 cmÀ1; HRMS (FAB): m/z calcd.
for C28H37NO12S: 611.2036 [M]; found: 611.2031.
Synthesis of thiophenyl(methyl-5-N-pentanoyl-4,7,8,9-tetra-OAc-
3,5-dideoxy-a-d-glycero-d-galacto-2-nonulopyranosylate)
(11):
The reaction was carried out according to the general procedure
with thioglycoside 8 (2.20 g, 3.77 mmol), methanesulfonic acid
(2.20 mL, 3.26 g, 33.9 mmol) and 2-methylpropionic acid anhydride
(0.040 mL, 0.038 mg, 6.79 mmol). Subsequent peracetylation was
achieved with acetic anhydride (5.40 mL, 5.83 g, 57.0 mmol) in pyri-
dine (11 mL). The reaction sequence delivered the product in
Synthesis of methyl 5-N-propanoyl-4,7,8,9-tetra-OAc-3,5-di-
deoxy-b-d-glycero-d-galacto-2-nonulopyranosylate (13): The re-
action was carried out according to the general procedure with thi-
ophenyl(methyl-5-N-propanoyl-4,7,8,9-tetra-OAc-3,5-dideoxy-a-d-
glycero-d-galacto-2-nonulopyranosylate) (9, 292 mg, 0.490 mmol)
and NBS (209 mg, 1.17 mmol) in acetone (10 mL) and distilled
water (0.7 mL). The product was obtained in a yield of 15%
(38 mg, 0.076 mmol) as a colourless syrup. [a]2D0 =À238 (c=1.0,
a yield of 95% (2.24 mg, 3.58 mmol) as a colourless syrup. [a]D20
=
=
1
3
À228 (c=0.90, CHCl3); H NMR (400 MHz, C6D6): d=7.54 (d, JH,H
8.3 Hz, 2H; Hortho), 7.18 (dd, 3JH,H =8.3, 8.0 Hz, 2H; Hmeta), 7.11 (d,
3JH,H =8.0 Hz,1H; Hpara), 5.55 (ddd, 3JH,H =7.2, 5.8, 2.6 Hz, 1H; H8),
5.45 (dd, 3JH,H =7.2, 2.3 Hz, 1H; H7), 4.92 (ddd, 3JH,H =12.1, 10.6,
CHCl3); H NMR (400 MHz, C6D6): d=5.72 (dd, JH,H =3.8, 2.4 Hz, 1H;
1
3
3
H7), 5.62 (ddd, 3JH,H =8.1, 3.8, 2.3 Hz, 1H; H8), 5.32 (ddd, JH,H
=
3
10.4, 10.4, 6.4 Hz, 1H; H4), 4.60 (ddd, JH,H =10.5, 10.4, 10.4 Hz, 1H;
2
3
3
2
4.7 Hz, 1H; H4), 4.60 (dd, JH,H =12.5 Hz, JH,H =2.6 Hz, 1H; H9), 4.54
(dd, 2JH,H =12.4 Hz, 3JH,H =5.8 Hz, 1H; H9), 4.32 (dd, 3JH,H =10.8,
10.6 Hz, 1H; H5), 4.12 (dd, JH,H =10.8, 2.3 Hz, 1H; H6), 3.33 (s, 3H;
H5), 4.40 (dd, JH,H =10.5, 2.4 Hz, 1H; H6), 4.25 (dd, JH,H =12.2 Hz,
3JH,H =8.1 Hz, 1H; H9), 3.32 (s, 3H; OCH3), 2.27–2.21 (m, 2H; H3ax/
H3eq), 2.01–1.84 (m, 2H; CH2-propanoyl), 1.94, 1.88, 1.71, 1.62 (4s,
3
OCH3), 2.84 (dd, 2JH,H =12.8 Hz, 3JH,H =4.7 Hz, 1H; H3eq), 2.15 (dd,
3JH,H =12.8, 2.1 Hz, 1H; H3ax), 2.07–2.05 (m, 2H; CH2-pentanoyl),
1.99, 1.84, 1.77, 1.70 (4s, 4ꢂ3H; 4CH3-acetyl), 1.47–1.38 (m, 2H;
CH2-pentanoyl), 1.14–1.05 (m, 2H; CH2-pentanoyl), 0.69 ppm (dd,
3JH,H =7.1, 7.1 Hz, 3H; CH3-pentanoyl); 13C NMR (101 MHz, C6D6):
d=175.0 (C1), 137.9 (Cortho), 131.2 (Cmeta), 129.8 (Cpara), 76.1 (C6),
72.0 (C8), 70.8 (C4), 68.7 (C7), 62.4 (C9), 52.2 (OCH3), 49.8 (C5), 39.6
(C3), 38.5 (C2-pentanoyl), 21.7, 21.2, 20.8, 20.7 (4CH3-acetyl), 19.1
(C3-pentanoyl), 14.9 (C4-pentanoyl), 13.1 ppm (C5-pentanoyl); IR:
4ꢂ3H; 4CH3-acetyl) 1.08 ppm (dd, JH,H =7.6, 7.6 Hz, 3H; CH3-prop-
3
anoyl); 13C NMR (101 MHz, C6D6): d=95.7 (C2), 73.5 (C8), 72.6 (C6),
69.7 (C4), 69.0 (C7), 63.5 (C9), 53.1 (OCH3), 49.7 (C5), 37.1 (C3), 30.0
(C2-propanoyl), 21.1, 20.9, 20.7, 20.7 (4CH3-acetyl), 10.1 ppm (C3-
propanoyl); IR: n˜ =3353, 2960, 1738, 1656, 1536, 1369, 1214, 1032,
603 cmÀ1
;
HRMS (FAB): m/z calcd. for C21H32NO13
:
506.1868
+
[M+H]+; found: 506.1896.
Synthesis of methyl 5-N-butanoyl-4,7,8,9-tetra-OAc-3,5-dideoxy-
b-d-glycero-d-galacto-2-nonulopyranosylate (14): The reaction
was carried out according to the general procedure with thiophe-
nyl(methyl-5-N-butanoyl-4,7,8,9-tetra-OAc-3,5-dideoxy-a-d-glycero-
d-galacto-2-nonulopyranosylate) (10, 170 mg, 0.280 mmol) and
NBS (128 mg, 0.720 mmol) in acetone (6.5 mL) and distilled water
(0.35 mL). The product was obtained in a yield of 64% (91 mg,
0.18 mmol) as a colourless solid. [a]2D0 =À298 (c=1.0, CHCl3);
1H NMR (400 MHz, C6D6): d=5.66 (dd, 3JH,H =4.3, 2.3 Hz, 1H; H7),
n˜ =3201, 3043, 2959, 1726, 1651, 1430, 1367, 1215, 1033, 751 cmÀ1
;
HRMS (FAB): m/z calcd. for C29H40NO12S+: 626.2 [M+H]+; found:
626.2.
Synthesis of thiophenyl(methyl-5-N-cyclopropylcarbonyl-4,7,8,9-
tetra-OAc-3,5-dideoxy-a-d-glycero-d-galacto-2-nonulopyranosy-
late (12): The reaction was carried out according to the general
procedure with thioglycoside 8 (1.00 g, 1.73 mmol), methanesul-
ChemBioChem 2012, 13, 2605 – 2615
ꢀ 2012 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
2611