Synthesis of histo blood-group antigens A and B (type 2), xenoantigen Galα1-3Galβ1-4GlcNAc and related type 2 backbone oligosaccharides as haptens in spacered form

Article abstract of DOI:10.1070/MC2002v012n04ABEH001591

The partial α-fucosylation of spacered (sp = CH₂CH₂CH₂NH₂) N-acetyl β-lactosamine derivative having free hydroxy groups at C-3 and C-2′ gives rise to oligosaccharides H (type 2), Le^x and Le^y, the first one was further elongated giving rise to A (type 2) or B (type 2) tetrasaccharides; spacered trisaccharides Galα1-3Galβ1-4GlcNAc and Galα1-4Galβ1-4GlcNAc were synthesised by the partial α-galactosylation of N-acetyl β-lactosamine derivative, having free hydroxy groups at C-4′ and C-3′.

Full text of DOI:10.1070/MC2002v012n04ABEH001591

Mendeleev Commun., 2002, 12(4), 143–145
Synthesis of histo blood-group antigens A and B (type 2), xenoantigen
Galα1-3Galβ1-4GlcNAc and related type 2 backbone oligosaccharides
as haptens in spacered form
Galina V. Pazynina, Tatiana V. Tyrtysh and Nicolai V. Bovin*
M. M. Shemyakin–Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow,
Russian Federation. Fax: +7 095 330 5592; e-mail: bovin@carb.siobc.ras.ru
10.1070/MC2002v012n04ABEH001591
The partial α-fucosylation of spacered (sp = CH2CH2CH2NH2) N-acetyl β-lactosamine derivative having free hydroxy groups at
C-3 and C-2' gives rise to oligosaccharides H (type 2), Lex and Ley, the first one was further elongated giving rise to A (type 2) or
B (type 2) tetrasaccharides; spacered trisaccharides Galα1-3Galβ1-4GlcNAc and Galα1-4Galβ1-4GlcNAc were synthesised by the
partial α-galactosylation of N-acetyl β-lactosamine derivative, having free hydroxy groups at C-4' and C-3'.
Type 2 blood group oligosaccharides, e.g., the Galβ1-4GlcNAc
OBn
O
OBn
backbone-based structures in the composition of glycoproteins
and glycolipids, are widespread antigens; namely, A, B, H and
P1 are major human erythrocyte and histo blood-group anti-
gens,1 trisaccharide B (type 2) is the main xenoantigen causing
hyperacute rejection during pig-to-human transplantation,2 Lex
(Lewis X) is CD15 antigen3 and Ley (Lewis Y) is one of the
most promising cancer-associated antigen for oncovaccination.4
Here we present a practical convergent synthesis of oligosac-
charides 1–7 in spacered form (R = CH2CH2CH2NH2) suitable
for the further synthesis of various glycoprobes.5 It should be
noted that the methods of glycosylation, as well as a protecting–
deprotecting strategy, are based on the approaches of H. Paulsen
and R. Lemieux with co-workers.6,7 In this work, we used a
very convenient aminopropyl spacer arm, as well as a really
practical strategy, which allowed us to synthesise several oligo-
saccharides by a single glycosylation procedure.
OR
O
O
O
BnO
Br–, Et2NPri
OH
O
Br
O
BnO
OBn
OH
NHAc
8
9
OBn
O
OBn
OBn
OBn
OR
OR
O
O
O
O
O
O
OH
O
O
O
O
O
O
BnO
NHAc
O
NHAc
O
O
BnO
BnO
BnO
OBn
BnO
BnO
GalNAcα1-3(Fucα1-2)Galβ1-4GlcNAcβ-OR A (type 2)
1
2
3
4
5
6
7
OBn
OBn
Galα1-3(Fucα1-2)Galβ1-4GlcNAcβ-OR
Fucα1-2Galβ1-4GlcNAcβ-OR
Galβ1-4(Fucα1-3)GlcNAcβ-OR
Fucα1-2Galβ1-4(Fucα1-3)GlcNAcβ-OR
Galα1-3Galβ1-4GlcNAcβ-OR
Galα1-4Galβ1-4GlcNAcβ-OR
(R = CH2CH2CH2NH2)
B (type 2)
H (type 2)
Lex
10
11
OBn
O
OBn
Ley
OR
O
O
O
Btri(type 2)
P1
R = CH2CH2CH2NHCOCF3
O
O
10
11
12
22
23
24
5
3
4
OH
NHAc
The synthesis of oligosaccharides 3–5 was accomplished as a
one-pot process based on a regio-nonspecific fucosylation of
lactosamine derivative 8 (the syntheses of this compound and
other starting compounds are described below) bearing two
hydroxyls, at C-3 of a glucosamine moiety and at C-2 of a
galactose moiety. The fucosylation of 87 with a sixfold excess
of fucosyl donor 9 (see Scheme 1) yields tetrasaccharide 10
(70–80%), whereas glycosylation with a smaller excess (2 mol)
of 9 provides an easy-to-separate (chromatography on silica gel)
mixture of tetrasaccharide 10 (19%) with trisaccharides 11 (43%)
and 12 (9%) and starting diol 8 (17%). Thus, limited glycosyla-
tion leads to a moderate yield of trisaccharide H derivative 11,
which is a building block for the synthesis of tetrasaccharides A
and B, together with ‘side’ oligosaccharides Lex 12 and Ley 10.
As shown in Scheme 2, derivative 11 of trisaccharide H (type
2) was converted into diol 13 (84–93% yield) by O-acetylation
followed by acid removal of benzylidene protection. Compound
13 was directly regioselectively α-galactosylated by bromide 14
at the 3-position giving rise to protected blood group B tetra-
saccharide 15 (50%) and side 1-4 tetrasaccharide (28%). Corre-
sponding A tetrasaccharide 16 was synthesised from alcohol 17
obtained from 13 by an orthoester procedure8 (89% yield, see
Scheme 2), using benzylated azido donor 18. The yield of tetra-
saccharide 16 was 40%; starting acceptor 17 (40%) was also
isolated from the reaction mixture. Alternatively, a derivative of
A tetrasaccharide 16a was obtained using diol 13 as a glycosyl
acceptor and bromide 18 as a glycosyl donor; in this case, the
yield was as low as 25%.
O
BnO
BnO
OBn
12
Scheme 1
Isomeric trisaccharides 20 and 21 were obtained in a one-pot
α-galactosylation procedure starting from another lactosamine
acceptor, diol 19, bearing hydroxyls at C-3 and C-4 positions of
a galactose moiety, and donor 14. The yields of major product
1-3 isomer 20 and minor 1-4 isomer 21 were 50 and 10%,
respectively (Scheme 3).
Fucosylation with donor 9 and galactosylation with donors
14 and 18, performed under standard conditions,6,7 were α-stereo-
selective; the corresponding β-anomeric products were isolated
by low pressure chromatography in minor (5–10%) amounts, if
any.
Standard deprotection6,7 of oligosaccharides 10–12, 15, 16,
20 and 21 gave spacered (sp = CH2CH2CH2NHCOCF3) oligo-
saccharides, which were purified and characterised by 1H NMR
spectra as per-O-acetates 22–249 and 25–28.† Acetylation was
performed using Ac2O/Py.
Starting disaccharide 8 was obtained by the glycosylation of
acceptor 2910 with 6-O-Bn galactosyl donor 3011 (68% yield)
followed by the Zemplen (catalytic amount of MeONa in MeOH)
deacetylation and acetonation at 3',4'-positions (Scheme 4), 78%
yield. Diol 19 (62%) was obtained from diol 8 by sequential
– 143 –

Mendeleev Commun., 2002, 12(4), 143–145
OBn
O
OBn
OR
OBn
O
OBn
OR
R'' O
HO
O
O
AgOTf
OBn
i, AcO/Py
ii, CF3COOH
OR'
OAc
OH
OAc
O
O
14
11
O
O
BnO
NHAc
OAc
NHAc
19
15
16
25
26
2
1
OBn
O
OBn
O
BnO
OBn
OR
HO
BnO
O
20
21
27
28
6
7
13 R = CH2CH2CH2NHCOCF3, R' = R'' = H
O
OAc
O
OBn
i, MeC(OEt)3, TsOH
ii, MeCOOH
17 R = CH2CH2CH2NHCOCF3, R' = H, R'' = Ac
OAc
NHAc
OBn
20
OBn
OBn
OBn
O
BnO
OBn
BnO
OBn
O
O
OBn
OBn
13
17
Br
Br
BnO
OR
O
O
O
OBn
OH
OAc
O
OBn
14
N3
18
OBn
OAc
NHAc
AgOTf
21
R = CH2CH2CH2NHCOCF3
OBn
Scheme 3
OBn
OBn
OR
BnO
OBn
O
O-acetylation and removal of isopropylidene protection,10 as
shown in Scheme 4.
R'' O
O
O
O
OAc
†
1H NMR spectra of oligosaccharide peracetates.
O
R'
25: Ac: 2.155 (s), 2.140 (2s), 2.106 (s), 2.086 (s), 2.068 (s), 2.065 (s),
2.061 (s), 1.981 (s), 1.953 (s); Galα1-3-: 5.322 (d, H-1, J1,2 3.5 Hz), 5.335
(dd, H-2), 5.40–5.45 (m, H-3), 5.598 (dd, H-4, J4,5 < 1.0 Hz), 4.446 (m,
H-5), 7.429 (m, NHCOCF3*); Fucα1-2-: 5.422 (d, H-1, J1,2 3.7 Hz),
5.207 (dd, H-2, J2,3 11.0 Hz), 5.040 (dd, H-3, J3,4 3.2 Hz), 5.248 (dd,
H-4, J4,5 1.0 Hz), 4.459 (m, H-5); >Galβ1-4-: 4.386 (d, H-1, J1,2 7.4 Hz),
3.553 (dd, H-2, J2,3 10.0 Hz), 3.880 (dd, H-3, J3,4 2.7 Hz), 5.406 (dd, H-4,
J4,5 < 1.0 Hz), 3.785 (m, H-5); -GlcNAcβ1-Osp: 4.494 (d, H-1, J1,2 7.6 Hz),
4.018 (ddd, H-2, J2,3 8.8 Hz), 5.066 (dd, H-3, J3,4 9.1 Hz), 3.911 (dd, H-4,
J4,5 8.3 Hz), 3.772 (m, H-5), 5.949 (d, NHAc, JNH,2 8.3 Hz).
O
NHAc
O
BnO
BnO
OBn
15 R = CH2CH2CH2NHCOCF3, R' = OBn, R'' = H
16 R = CH CH CH NHCOCF , R = N , R = Ac
'
''
2
2
2
3
3
26: Ac: 1.906 (s), 1.965 (s), 1.982 (s), 1.993 (s), 2.066 (3s), 2.076 (s),
2.101 (s), 2.105 (s), 2.146 (s), 2.157 (s); Fuc Me: 1.15 (d, 3H, J5,6 6.6 Hz);
GalNAcα1-3-: 5.212 (d, H-1, J1,2 3.20 Hz), 4.450 (m, H-2), 5.003 (dd,
H-3, J3,4 3.0 Hz), 5.420 (dd, H-4, J4,5 < 1.0), 6.316 (d, NHAc, JNH,2
9.4 Hz), 7.444 (m, NHCOCF3*); Fucα1-2-: 5.508 (d, H-1, J1,2 3.5 Hz),
5.274 (dd, H-2, J2,3 11.0 Hz), 5.104 (dd, H-3, J3,4 3.0 Hz), 5.300 (dd, H-4,
J4,5 < 1.0 Hz); >Galβ1-4-: 4.403 (d, H-1, J1,2 7.3 Hz), 3.748 (dd, H-2,
J2,3 9.5 Hz), 3.868 (dd, H-3, J3,4 3.5 Hz), 5.372 (dd, H-4, J4,5 < 1.0 Hz);
–GlcNAcβ1-Osp: 4.487 (d, H-1, J1,2 8.0 Hz), 3.990 (ddd, H-2, J2,3 9.0 Hz),
5.072 (dd, H-3, J3,4 10.0 Hz), 3.860 (dd, H-4, J4,5 8.5 Hz), 5.886 (d,
NHAc, JNH,2 8.9 Hz).
27 (300 MHz): Ac: 1.951 (s), 1.978 (2s), 1.998 (s), 2.062 (s), 2.079 (s),
2.085 (s), 2.118 (s), 2.125 (s), 2.139 (2s), 2.159 (s); Fuc Me: 1.168 (d, 3-H,
J5,6 7.0 Hz); Galα1-3-: 5.254 (d, H-1, J1,2 3.5 Hz), 5.269 (dd, H-2, J2,3
10.7 Hz), 5.112 (dd, H-3, J3,4 3.1 Hz), 5.453 (dd, H-4, J4,5 < 1.0 Hz),
4.223 (m, H-5), 7.399 (m, NHCOCF3*); –Galβ1-4-: 4.454 (d, H-1, J1,2
8.0 Hz), 5.164 (dd, H-2, J2,3 10.2 Hz), 3.863 (dd, H-3, J3,4 3.0 Hz), 5.347
(dd, H-4, J4,5 < 1.0 Hz), 3.821 (m, H-5); –GlcNAcβ1-Osp: 4.418 (d, H-1,
J1,2 7.06 Hz), 4.029 (ddd, H-2, J2,3 9.0 Hz), 5.061 (dd, H-3, J3,4 8.6 Hz),
3.775 (dd, H-4, J4,5 8.1 Hz), 3.60–3.68 (m, H-5), 5.909 (d, NHAc, JNH,2
8.9 Hz).
28 (300 MHz): Ac: 1.942 (s), 1.957 (s), 2.017 (s), 2.022 (s), 2.044 (2s),
2.054 (s), 2.079 (2s), 2.101 (s); Galα1-4-: 4.968 (d, H-1, J1,2 3.5 Hz);
5.167 (dd, H-2, J2,3 11.0 Hz), 5.347 (dd, H-3, J3,4 3.3 Hz), 5.542 (dd, H-4,
J4,5 < 1.0 Hz), 4.450 (m, H-5), 7.553 (m, NHCOCF3*); –Galβ1-4-: 4.534
(d, H-1, J1,2 7.5 Hz), 5.083 (dd, H-2, J2,3 10.5 Hz), 4.740 (dd, H-3, J3,4
2.4 Hz), 4.011 (dd, H-4, J4,5 < 1.0 Hz), 3.756 (m, H-5); –GlcNAcβ1-Osp:
4.411 (d, H-1, J1,2 7.3 Hz), 4.000 (ddd, H-2, J2,3 9.0 Hz), 5.049 (dd, H-3,
J3,4 9.4 Hz), 3.756 (dd, H-4, J4,5 8.6 Hz), 3.620 (m, H-5), 6.118 (d, NHAc,
JNH,2 8.8 Hz).
16a R = CH CH CH NHCOCF , R = N , R = H
'
''
2
2
2
3
3
Scheme 2
OBn
OR
OBn
O
i, AgOTf
ii, MeONa/MeOH
iii, Me2C(OMe)2, TsOH
AcO
O
OAc
OAc
Br
8
i, Ac2O/Py
ii, CF3COOH
HO
NHAc
29
OAc
30
19
Scheme 4
The Zemplen de-O-acetylation of 22–28 followed by removal
of the N-trifluoroacetyl group with Amberlyst (in OH– form) gave
oligosaccharides 5,9 3, 4,9 2, 1, 6 and 7, respectively, bearing
(CH2)3NH2 spacer groups. The NMR data of compounds 1–7
are consistent with the assumed structures: J1,2 values corre-
spond to expected anomeric configurations; characteristic groups
such as Me–C and MeCONH are unambiguously identified.‡
This work was supported in part by NIH (grant no.
1U54GM62116).
References
1 H. Clausen and S. Hakomori, Vox Sang., 1989, 56, 1.
2 D. K. C. Cooper, Xenotransplantation, 1998, 5, 6.
* Signals of O(CH2)3N: 1.75–1.95, 3.20–3.35, 3.55–3.70, 3.85–3.095,
H-5, Hb-6a; H-6b: 3.50–4.50.
– 144 –

Mendeleev Commun., 2002, 12(4), 143–145
3 Leukocyte Typing VI, ed. T. Kishimoto, Garland Publishing, New York,
1997, p. 1121.
4 S. Zhang, H. S. Zhang, C. Cordon-Cardo, V. E. Reuter, A. K. Singhal,
K. O. Lloyd and P. O. Livingston, Int. J. Cancer, 1997, 73, 50.
5 N. V. Bovin, Glycoconjugate J., 1998, 15, 431.
6
7
8
9
H. Paulsen, in Organic Synthesis: An Interdisciplinary Challenge, eds.
J. Streith, H. Prinzbach and G. Schill, Blackwell, Oxford, 1985, p. 317.
O. Hindsgaul, T. Norberg, J. le Pendu and R. U. Lemieux, Carbohydr.
Res., 1982, 109, 109.
A. Ya. Chernyak, A. B. Levinsky, B. A. Dmitriev and N. K. Kochetkov,
Carbohydr. Res., 1984, 128, 269.
‡
1
1, Atetr (type 2): H NMR (500 MHz, D2O) d: 5.280 (d, 1H, H-1, J1,2
Detailed NMR data of the H (type 2), Ley and Lex in spacered form,
obtained by alternative approach based on 1,6-anhydro-GlcNAc deriva-
tive, are published in: T. V. Tyrtysh, N. E. Byramova, A. B. Tuzikov,
and N. V. Bovin, Bioorg. Khim., 2002, in press.
3.9 Hz), 5.108 (d, 1H, H-1, J1,2 3.7 Hz), 4.526 (d, 1H, H-1, J1,2 7.6 Hz),
4.432 (d, 1H, H-1, J1,2 8.6 Hz), 1.982 (s, 3H, NHCOMe), 1.970 (s, 3H,
NHCOMe), 1.175 (d, 3H, Me fucose, J5,6 6.6 Hz). MS, m/z: 812
(789 + 23, M+ + Na+), [a]D +17 (c1, H2O).
10 N. E. Nifant’ev, Y. E. Tsvetkov, A. S. Shashkov, L. O. Kononov, V. M.
Menshov, A. B. Tuzikov and N. V. Bovin, J. Carbohydr. Chem., 1996,
15, 939.
11 N. E. Byramova, E. Yu. Korchagina, T. V. Ovchinnikova, A. B. Tuzikov,
G. V. Pazynina, H.-J. Gabius and N. V. Bovin, Bioorg. Khim., 2002, in
press.
1
2, Btetr (type 2): H NMR (500 MHz, D2O) d: 5.303 (d, 1H, H-1, J1,2
4.2 Hz), 5.216 (s, 1H, H-1), 4.587 (d, 1H, H-1, J1,2 7.6 Hz), 4.464 (d,
1H, H-1, J1,2 8.3 Hz), 2.027 (s, 3H, NHCOMe), 1.208 (d, 3H, Me fucose,
J5,6 6.6 Hz). 13C NMR (500 MHz, D2O) d: 102.54 (C-1), 101.54 (C-1),
100.08 (C-1), 94.39 (C-1), 56.60 (C-2, CNHAc). MS, m/z: 772 (749 + 23,
M+ + Na+). [a]D –5 (c1, H2O).
1
6, Btri (type 2): H NMR (500 MHz, D2O) d: 5.127 (d, 1H, H-1, J1,2
3.7 Hz), 4.522 (d, 1H, H-1, J1,2 7.9 Hz), 4.501 (d, 1H, H-1, J1,2 7.9 Hz),
2.031 (d, 3H, NHCOMe). 13C NMR (300 MHz, D2O) d: 104.03 (C-1),
102.37 (C-1), 96.67 (C-1), 56.25 (C-2, CNHAc). MS, m/z: 626 (603 + 23,
M+ + Na+). [a]D +54 (c1, H2O).
1
7, P1: H NMR (500 MHz, D2O) d: 4.918 (d, 1H, H-1, J1,2 3.9 Hz),
4.497 (d, 1H, H-1, J1,2 7.6 Hz), 4.485 (d, 1H, H-1, J1,2 7.6 Hz), 2.019 (s,
3H, NHCOMe). MS, m/z: 626 (603 + 23, M+ + Na+). [a]D +30 (c1, H2O).
Received: 16th April 2002; Com. 02/1917
– 145 –