Block synthesis of blood group tetrasaccharides B (types 1, 3 and 4)

Article abstract of DOI:10.1016/j.mencom.2009.05.013

3-Aminopropyl glycosides of the tetrasaccharides Galα1-3(Fucα1-2)Galβ1-3GlcNAcβ (B type 1), Galα1-3(Fucα1-2)-Galβ1-3GalNAcα (B type 3), and Galα1-3(Fucα1-2)Galβ1-3GalNAcβ (B type 4) were synthesised using acetylated Galα1-3-(Fucα1-2)Gal trichloroacetimida

Full text of DOI:10.1016/j.mencom.2009.05.013

Mendeleev
Communications
Mendeleev Commun., 2009, 19, 152–154
Block synthesis of blood group
tetrasaccharides B (types 1, 3 and 4)
Elena Yu. Korchagina,a Ivan M. Ryzhov,a Konstantin A. Byrgazov,a
Inna S. Popova,a Sergei N. Pokrovskyb and Nicolai V. Bovin*a
a
M. M. Shemyakin–Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences,
117997 Moscow, Russian Federation. Fax: +7 495 330 5592; e-mail: bovin@carb.ibch.ru
b Institute of Experimental Cardiology, Cardiology Research Center, 121552 Moscow, Russian Federation
DOI: 10.1016/j.mencom.2009.05.013
3-Aminopropyl glycosides of the tetrasaccharides Galα1-3(Fucα1-2)Galβ1-3GlcNAcβ (B type 1), Galα1-3(Fucα1-2)-
Galβ1-3GalNAcα (B type 3), and Galα1-3(Fucα1-2)Galβ1-3GalNAcβ (B type 4) were synthesised using acetylated Galα1-3-
(Fucα1-2)Gal trichloroacetimidate 4 as a glycosyl donor at the key stage.
Originally discovered on red cells, the blood group ABO(H)
antigens are secreted in saliva and expressed in many human
tissues, especially, on endothelial and epithelial cells.1 They
are responsible for hemolytic reactions in blood transfusion
and acute rejection in bone marrow and organ transplantation.
Their expression varies during cell differentiation, maturation and
malignant transformations in cancer and cardio-vascular diseases.2
Fundamental studies in glycobiology and, moreover, the solution
of problems in practical hematology and transplantology require
oligosaccharides determining the blood group antigen specificity
in amounts that cannot be obtained from natural sources. Thus,
the access to pure blood group oligosaccharides fully relies on
OH
OH
OH
OH
O
O
OH
OH
OH
OH
OH
O
OH
O
OH
O
HO
HO
HO
O
O
HO
HO
O
O
O
O
NH2
O
NHAc
O
AcHN
NH2
O
O
O
OH
OH
OH
OH
HO
HO
1 B (type 1)
2 B (type 3)
Ph
OH
OAc
OH
O
OAc
O
O
OH
OH
OAc
OH
O
OH
O
O
NHAc
O
HO
OAc
O
O
HO
AcO
X
HO
AcO
O
O
NHAc
O
NH2
5
O
O
CCl3
NH
O
X = O(CH2)3NHCOCF3
O
O
OH
OAc
OH
OAc
HO
AcO
Ph
4
3 B (type 4)
O
O
OR
OR
O
v, vi
O
OAc
OR
OAc
O
OR
O
HO
X
RO
OAc
N3
RO
OAc
O
AcO
iv
O
X
Y
AcO
O
O
6 X = H, Y = O(CH2)3NHCOCF3
7 X = O(CH2)3NHCOCF3, Y = H
OAc
OAc
O
9
O
OR
OR
O
RO
OAc
8 R = H, X = O(CH2)3NH2
9 R = Ac, X = O(CH2)2
AcO
But
10
i, ii, iii
O
N
i, ii, iii
But
9a R = Ac, X = O(CH2)2CHO
Scheme 1 Synthesis of trichloroacetimidates of acetylated trisaccharide B. Reagents and conditions: i, 3,5-di-tert-butyl-1,2-benzoquinone, MeOH;
ii, H2C2O4; iii, Ac2O/Py; iv, AcOH–Ac2O–AcONa, 100 °C; v, N2H4·AcOH/DMF, 50 °C; vi, Cl3CCN, DBU, CH2Cl2, –20 °C ® room temperature.
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© 2009 Mendeleev Communications. All rights reserved.

Mendeleev Commun., 2009, 19, 152–154
OAc
OAc
OAc
OAc
O
Ph
O
OAc
OAc
OAc
O
sp
NHAc
OAc
O
OAc
O
AcO
i
AcO
O
O
O
AcO
AcO
AcO
sp
NHAc
O
O
O
O
iii, iv
80%
vi, vii
90%
1
4 + 5
O
O
O
O
OAc
11
OAc
OAc
OAc
AcO
AcO
12
OAc
OAc
O
sp = O(CH2)3NHCOCF3
OAc
OAc
O
AcO
Ph
AcO
O
O
O
O
O
sp
NHAc
O
O
OAc
11a
OAc
AcO
Ph
OAc
OAc
OAc
O
OAc
O
O
OAc
OAc
AcO
O
O
OAc
O
OAc
O
OAc
O
AcO
AcO
O
AcO
AcO
X
X
O
O
O
iii, iv, v
vi, vii
90%
ii
4 + 6 (7)
N3
Y
2 (3)
AcHN
O
O
Y
53% (40%)
O
O
OAc
13, 14
OAc
15, 16
OAc
OAc
AcO
AcO
OAc
OAc
O
Ph
OAc
OAc
O
AcO
O
AcO
O
O
O
O
X
O
O
N3
OAc
Y
OAc
AcO
13, 13a, 15 X = H, Y = O(CH2)3NHCOCF3
14, 14a, 16 X = O(CH2)3NHCOCF3, Y = H
13a, 14a
Scheme 2 Synthesis of aminopropyl glycosides of B tetrasaccharides (types 1, 3 and 4). Reagents and conditions: i, TMSOTf, MeCN–CH2Cl2 (1:1), MS
4 Å; ii, TMSOTf, MeCN, MS 4 Å; iii, 80% AcOH, 80 °C; iv, Ac2O/Py; v, Ph3P, THF/H2O, Ac2O/Py; vi, MeONa, MeOH; vii, NaOH, H2O.
chemical3–8 and chemoenzymatic9,10 syntheses. Here, we report
a chemical block-synthetic approach to obtain blood group B
tetrasaccharides of various types, which allows us to minimize
the number of synthetic steps.
benzoxazole derivative 9 in 88% yield. The structure of com-
pound 9 was confirmed by mass spectrometry [m/z 1124
(M+ + H), 1146 (M+ + Na)] and 1H NMR spectroscopy [two
siglets at d 1.44 (9H) and 1.49 (9H) related to tert-butyl groups
and a multiplet at d 7.2–7.5 (2H) of aromatic protons] data.
Base-catalyzed acetolysis/acetylation14 of benzoxazole 9 gave
peracetylated trisaccharide 10 in 90% yield. Then, selective
1-O-deacetylation of derivative 10 with hydrazine acetate16 and
treating with trichloroacetonitrile in the presence of 1,8-diazo-
bicyclo[5.4.0]undec-7-ene (DBU) gave glycosyl trichloro-
acetimidate 4, which was isolated by column chromatography
on silica gel [elution: hexane–ethyl acetate (1:1), 1% triethyl-
amine] as a mixture of α/β anomers in 80% yield.
Glycosylation of derivatives 5, 6, and 7 (Scheme 2) with
trichloroacetimidate 4 was the key stage in tetrasaccharide
synthesis. The glycosylation was carried out in acetonitrile (or
its mixture with dichloromethane) at room temperature using
a two-fold excess of the glycosyl acceptor and trimethylsilyl
triflate (0.1 equiv.) as the reaction promoter.
Tetrasaccharides 11, 13, and 14 were isolated by column
chromatography on silica gel in 52, 60, and 55% yields, respec-
tively. The β-configuration of generated glycosidic bond for com-
pounds 11, 13, and 14 was confirmed by 1H NMR spectroscopy
including spin decoupling and COSY experiments.‡ α-Isomers
of tetrasaccharides type 1 (11a) and type 4 (14a) were isolated
in 4 and 10% yields, respectively. We were not able to isolate
α-isomer of tetrasaccharide type 3 present in the mixture in
The methodology of stepwise elongation of the carbohydrate
chain starting from the reducing end is most often used in the
synthesis of blood group tetrasaccharides.3–7 Only in one paper8
it is reported that the disaccharide glycosyl donor has been used
for the synthesis of blood group tetrasaccharide A (type 3).
Blood group tetrasaccharides B 1–3† contain the same structural
fragment, trisaccharide B. This allows one to use the block
scheme ‘3 + 1’ in their synthesis, where ‘3’ is a glycosyl donor,
in particular, acetylated glycosyl trichloroacetimidate of trisac-
charide 4 and ‘1’ is the corresponding glucosamine (511) or
galactosamine (612 or 712) glycosyl acceptor. As far as we know,
2,3-branched oligosaccharides have not been used as glycosyl
donors for synthesis of 1,2-trans-glycosides.
Aminopropyl glycoside 813 has been selected as a precursor
for the preparation of the glycoside donor, and the method of
aminopropyl spacer elimination published previously14 has been
optimized (Scheme 1). In the cited paper,14 aldehyde 9a was
obtained in a low total yield (41%) by derivatization of amino-
propyl glycoside 5 with 3,5-di-tert-butyl-1,2-benzoquinone15 (BQ),
treatment of the resulting azomethine with oxalic acid dihydrate,
and acetylation. We have changed the ratio of reagents in
azomethine synthesis [8:BQ, 1:1.5 (1:114)] and increased the
pH value upon acidification from pH 214 to pH 4, thus affording
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Mendeleev Commun., 2009, 19, 152–154
negligible amount. The high stereoselectivity of glycosylation
purified by ion-exchange chromatography on DOWEX (H+)
cationite. The structure of the compounds synthesized was con-
firmed by 1H NMR (COSY) spectroscopy and mass spectrometry.
Thus, the suggested block scheme is advantageous for the
synthesis of blood group B tetrasaccharides. We suppose to use
this approach in the further syntheses of the more complex blood
group oligosaccharides, in particular, biantennary structures,
containing two fragments of B trisaccharide in one molecule.
can be explained primarily by the glycosyl donor structure. The
bulky substituent at C-2, acetylated fucose, and conformational
rigidity of the molecule due to the presence of two monosac-
charide residues in adjacent positions (Fuc at C-2 and Gal at
C-3) hinder the nucleophilic attack from α-side, thus resulting
in the formation of β-glycosides. Additionally, the use of aceto-
nitrile as the solvent upon glycosylation also affects positively
the reaction stereochemistry.17
The target aminopropyl glycosides of tetrasaccharides 1–3
were obtained by conventional deprotection methods (Scheme 2).
The protected oligosaccharides were isolated by column chromato-
graphy on silica gel, whereas aminopropyl glycosides 1–3 were
This work was supported by the Russian Foundation for Basic
Research (grant no. 07-04-00630), RAS Presidium Programme
for Molecular and Cell Biology, and Federal Target Programme
for Live Systems (state contract no. 02.512.11.2100).
†
This paper describes the synthesis of three tetrasaccharides, in which
Online Supplementary Materials
Supplementary data associated with this article can be found
in the online version at doi:10.1016/j.mencom.2009.05.013.
B-trisaccharide is linked to the hydroxyl group at C-3 of glucosamine
(type 1) or galactosamine (type 3 and type 4) via the β-glycosidic bond.
Synthesis of tetrasaccharide B (type 2), where B-trisaccharide is linked
to the 4-OH group of glucosamine, will be described elsewhere.
Spectral characteristics for oligosaccharides. 1H NMR spectra were
recorded at 30 °C on Bruker WM 500 MHz and Bruker WM 600 MHz
instruments. Mass spectra (MALDI-TOF) were recorded on a VISION 2000
mass spectrometer. Optical rotations were measured on a Perkin Elmer 341
polarimeter at 25 °C.
References
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R. Oriol, R. Mollicone, P. Couillin, A.-M. Dalix and J.-J. Candelier,
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Symbols of monosaccharide residues for trisaccharides: a – α/β-Gal
(reducing end), b – α-Gal, c – α-Fuc, for tetrasaccharides: a – α/β-GalNAc
or β-GlcNAc, b – β-Gal, c – α-Gal, d – α-Fuc.
3
4
1: 1H NMR (selected data, 600 MHz, 40 °C, D2O) d: 1.37 (d, 3H, H-6d,
J5,6 6.6 Hz), 2.05–2.12 (m, 2H, CCH2C), 2.21 (s, 3H, Ac), 3.19–3.23 (m,
2H, NHCH2), 4.47–4.50 (m, 1H, H-5d), 4.56 (d, 1H, H-1a, J1,2 8.4 Hz),
4.84 (d, 1H, H-1b, J1,2 7.8 Hz), 5.36 (d, 1H, H-1c, J1,2 5.0 Hz), 5.37 (d,
1H, H-1d, J1,2 4.6 Hz). MS, m/z: 749 (M+ + H), 771 (M+ + Na), 787
(M+ + K). [a]D –33.2 (c 1, H2O).
5
6
7
8
9
2: 1H NMR (selected data, 500 MHz, D2O) d: 1.20 (d, 3H, H-6d,
J5,6 6.4 Hz), 1.95–2.02 (m, 2H, CCH2C), 2.06 (s, 3H, Ac), 3.04–3.15 (m,
2H, NHCH2), 3.49–3.56 (m, 1H, OCHH), 4.70 (d, 1H, H-1b, J1,2 7.4 Hz),
4.90 (d, 1H, H-1a, J1,2 3.0 Hz), 5.25 (d, 1H, H-1c, J1,2 2.6 Hz), 5.27 (d,
1H, H-1d, J1,2 4.2 Hz). MS, m/z: 749 (M+ + H), 771 (M+ + Na), 787
(M+ + K). [a]D +81.0 (c 0.5, H2O).
W. M. Macindoe, H. Ijima, Y. Nakahara and T. Ogawa, Carbohydr.
Res., 1995, 269, 227.
C. A. G. M. Weijers, M. C. R. Franssen and G. M. Visser, Biotechnol.
Adv., 2008, 26, 436.
10 O. Blixt and N. Razi, Methods Enzymol., 2006, 415, 137.
11 N. V. Bovin, T. V. Zemlyanukhina, C. N. Chagiashvili and A. Ya. Khorlin,
Khim. Prir. Soedin., 1988, 6, 777 [Chem. Nat. Compd. (Engl. Transl.),
1988, 6, 659].
12 T. V. Ovchinnikova, A. G. Ter-Grigorian, G. V. Pazynina and N. V.
Bovin, Bioorg. Khim., 1997, 23, 61 (Russ. J. Bioorg. Chem., 1997, 23, 55).
13 E. Yu. Korchagina and N. V. Bovin, Bioorg. Khim., 1992, 18, 283
(Russ. J. Bioorg. Chem., 1992, 18, 153).
3: 1H NMR (selected data, 600 MHz, D2O) d: 1.24 (d, 3H, H-6d,
J5,6 6.4 Hz), 1.92–2.02 (m, 2H, CCH2C), 2.09 (s, 3H, Ac), 3.10–3.13 (m,
2H, NCH2), 4.36 (d, 1H, H-1a, J1,2 7.9 Hz), 4.72 (d, 1H, H-1b, J1,2 7.6 Hz),
5.27 (d, 1H, H-1c, J1,2 3.1 Hz), 5.29 (d, 1H, H-1d, J1,2 4.1 Hz). MS, m/z:
749 (M+ + H), 771 (M+ + Na), 787 (M+ + K). [a]D +5.0 (c 0.5, H2O).
‡
11: 1H NMR (500 MHz, CDCl3) d: 1.30 (d, 3H, H-6d, J5,6 6.4 Hz),
14 E. V. Shipova and N. V. Bovin, Mendeleev Commun., 2000, 63.
15 E. J. Corey and K. Achiwa, J. Am. Chem. Soc., 1969, 9, 1429.
16 G. Excoffier and D. Gagnaire, Carbohydr. Res., 1975, 39, 368.
17 R. R. Schmidt and W. Kinsy, Adv. Carbohydr. Chem. Biochem., 1994,
50, 21.
1.90, 1.95, 1.98, 2.01, 2.06, 2.08, 2.14, 2.16, 2.17, 2.19 (10s, 10×3H, 10Ac),
3.14–3.18 (m, 1H, H-2a), 3.17–3.25 (m, 1H, NHCHH), 3.43–3.50 (m,
1H, OCHH), 3.56 (dd, 1H, H-2b, J1,2 7.7 Hz, J2,3 10.0 Hz), 3.59–3.64
(m, 1H, H-5a), 3.63–3.70 (m, 1H, NHCHH), 3.70 (dd, 1H, H-3b, J2,3 10.0 Hz,
J3,4 2.8 Hz), 3.78–3.87 (m, 3H, H-3a, H-4a, H-5a), 3.88–3.92 (m, 1H,
H-5b), 3.95–4.04 (m, 2H, H-6'c, OCHH), 4.05–4.13 (m, 2H, H-6'a, H-6'b),
4.28–4.36 (m, 2H, H-6c, H-6b), 4.37 (d, 1H, H-1a, J1,2 8.5 Hz), 4.45–4.49
(m, 1H, H-5c), 4.53 (d, 1H, H-1b, J1,2 7.7 Hz), 4.57–4.62 (m, 1H, H-5d),
4.98–5.12 (m, 2H, H-2d, H-3d), 5.24 (dd, 1H, H-4b, J3,4 2.8 Hz,
J4,5 < 1 Hz), 5.30 (dd » br. s, 1H, H-4d), 5.34 (d, 1H, H-1c, J1,2 3.8 Hz),
5.36 (dd, 1H, H-2c, J1,2 3.8 Hz, J2,3 10.9 Hz), 5.45 (dd, 1H, H-3c,
J2,3 10.9 Hz, J4,3 3.2 Hz), 5.46 (d, 1H, H-1d, J1,2 2.9 Hz), 5.52 (s, 1H,
PhCH), 5.58 (dd, 1H, H-4c, J3,4 3.2 Hz, J4,5 < 1 Hz), 7.02 (d, 1H, NHAc,
JNH,2 7.6 Hz), 7.35–7.48 (m, 5H, Ph), 7.91–7.96 [m, 1H, NHC(O)CF3].
13: 1H NMR (500 MHz, CDCl3) d: 0.98 (d, 3H, H-6d, J5,6 6.6 Hz),
1.93, 1.99, 2.00, 2.04, 2.08, 2.09, 2.12, 2.14, 2.15 (9s, 9×3H, 9Ac),
3.49–3.65 (m, 3H, NHCH2, OCHH), 3.70–3.74 (m, 1H, H-5a), 3.80–3.85
(m, 2H, H-2b, H-5b), 3.90–3.97 (m, 2H, H-3b, OCHH), 4.06–4.29 (m,
8H, H-2a, H-3a, H-6a, H-6'a, H-6b, H-6'b, H-6c, H-6'c), 4.39 (dd, 1H,
H-4a, J3,4 2.9 Hz, J4,5 < 1 Hz), 4.41–4.45 (m, 1H, H-5c), 4.45–4.51
(m, 1H, H-5d), 4.83 (d, 1H, H-1b, J1,2 7.2 Hz), 5.11 (d, 1H, H-1a,
J1,2 3.4 Hz), 5.19–5.26 (m, 3H, H-2d, H-3d, H-4d), 5.30 (dd, 1H, H-2c,
J1,2 3.6 Hz, J2,3 10.9 Hz), 5.36 (d, 1H, H-1c, J1,2 3.6 Hz), 5.38 (dd, 1H,
H-3c, J2,3 10.9 Hz, J3,4 3.0 Hz), 5.47 (dd » br. s, 1H, H-4b), 5.58 (dd,
1H, H-4c, J3,4 3.0 Hz, J4,5 < 1 Hz), 5.63 (s, 1H, CHPh), 5.65 (d, 1H, H-1d,
J1,2 3.4 Hz), 6.89–6.93 [m, 1H, NHC(O)CF3], 7.31–7.58 (m, 5H, Ph).
MS, m/z: 1317 (M+ + Na), 1333 (M+ + K). [a]D +75.2 (c 1, CHCl3).
Received: 25th November 2008; Com. 08/3241
14: 1H NMR (500 MHz, CDCl3) d: 0.96 (d, 3H, H-6d, J5,6 6.4 Hz),
1.91–1.97 (m, 2H, CCH2C), 1.92, 1.98, 1.99, 2.05, 2.06, 2.07, 2.10, 2.14,
2.15 (9s, 9×3H, 9Ac), 3.43–3.45 (m, 1H, H-5a), 3.46–3.62 (m, 2H,
NHCH2), 3.69 (dd, 1H, H-3a, J2,3 10.5 Hz, J3,4 3.4 Hz), 3.71–3.75
(m, 1H, OCHH), 3.76–3.81 (m, 2H, H-2b, H-5b), 3.90 (dd, 1H, H-3b,
J2,3 9.7 Hz, J3,4 3.0 Hz), 4.01–4.06 (m, 1H, OCHH), 4.07–4.19 (m, 6H,
H-2a, H-6'a, H-6b, H-6'b, H-6c, H-6'c), 4.25 (dd, H-4a, J3,4 3.4 Hz,
J4,5 < 1 Hz), 4.28–4.32 (m, 1H, H-6a), 4.38–4.47 (m, 3H, H-1a, H-5c,
H-5d), 5.01 (d, 1H, H-1b, J1,2 7.4 Hz), 5.18 (dd, 1H, H-3d, J2,3 10.9 Hz,
J3,4 3.2 Hz), 5.22 (dd, 1H, H-4d, J3,4 3.2 Hz, J4,5 < 1 Hz), 5.25 (dd, 1H,
H-2d, J1,2 3.6 Hz, J2,3 10.9 Hz), 5.28 (dd, 1H, H-2c, J1,2 3.2 Hz, J2,3 10.7 Hz),
5.34 (d, 1H, H-1c, J1,2 3.2 Hz), 5.37 (dd, 1H, H-3c, J2,3 10.7 Hz, J3,4 3.1 Hz),
5.43 (dd, 1H, H-4b, J3,4 3.0 Hz, J4,5 < 1 Hz), 5.57 (dd, 1H, H-4c, J3,4 3.1 Hz,
J4,5 < 1 Hz), 5.60 (s, 1H, CHPh), 5.64 (d, 1H, H-1d, J1,2 3.6 Hz), 7.02–7.08
[m, 1H, NHC(O)CF3], 7.33–7.54 (m, 5H, Ph). MS, m/z: 1317 (M+ + Na),
1333 (M+ + K). [a]D +1.2 (c 1, CHCl3).
For spectral characteristics of compounds 4, 9, 10-α, 10-β, 12, 15 and
16, see Online Supplementary Materials.
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