Synthesis of some divalent O- and S-glycosides of galabiose and globotriose

Article abstract of DOI:10.1016/S0008-6215(98)00021-4

Derivatives of galabiose (α-D-Galp-(1→4)-d-Galp) and globotriose (a- D-Galp-(1→4)-β-D-Galp(l→4)-D-Glcp) were coupled to various 1,2- and 1,3- dihydroxymethyl- and dimercaptomethylbenzenes to give the corresponding divalent glycosides, potentially useful as inhibitors of bacterial adhesion.

Full text of DOI:10.1016/S0008-6215(98)00021-4

Carbohydrate Research 307 (1998) 243±251
Synthesis of some divalent O- and S-glycosides
of galabiose and globotriose
Henrik C. Hansen, Goran Magnusson*
Organic Chemistry 2, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124,
S-221 00 Lund, Sweden
Received 2 October 1997; accepted 20 December 1997
Abstract
Derivatives of galabiose (ꢀ-d-Galp-(1!4)-d-Galp) and globotriose (a-d-Galp-(1!4)-ꢁ-d-Galp-
(1!4)-d-Glcp) were coupled to various 1,2- and 1,3-dihydroxymethyl- and dimercapto-
methylbenzenes to give the corresponding divalent glycosides, potentially useful as inhibitors of
bacterial adhesion. # 1998 Elsevier Science Ltd. All rights reserved
Keywords: Divalent galabiosides; Divalent globotriosides, synthesis of
1. Introduction
described recently that nanomolar concentrations
of glycodendrimers carrying two to four galabiosyl
residues can inhibit hemagglutination between the
pig pathogen Streptococcus suis and red blood cells
[5]. The inhibitory eciency of the glycoden-
drimers depends not only on the number of sac-
charides present in the molecule, but also on the
mode of presentation of the binding epitopes. In
line with an attempt to obtain additional informa-
tion about the binding phenomena, we now report
the synthesis of a number of novel divalent glyco-
sides carrying the galabiose and globotriose sac-
charide moieties.
Glycolipids containing the disaccharide moiety
galabiose [Gal(ꢀ1-4)Gal] function as receptors for
pathogen adhesion to cells, which constitutes the
®rst step of an infective process [1]. The biological
background to these phenomena has been sum-
marized in several publications from this labora-
tory [2]. Blocking of the carbohydrate±protein
recognition by receptor analogs is a potentially
useful approach towards novel antibacterial
agents, similar to the recent development of anti-
adhesive antiviral compounds [3].
Most carbohydrate±protein interactions are
rather weak and millimolar concentrations of an
inhibitory saccharide derivative are often required
for complete inhibition. Weak interactions can be
compensated by the use of multivalent inhibitors,
as demonstrated by several research groups [4]. We
2. Results and discussion
A series of commercially available divalent
benzene derivatives were chosen as sca€olds for
the construction of the divalent glycosides. Thus,
1,2-dimercaptomethylbenzene (Scheme 1) was gly-
cosylated with the known [5] galabiose imidate 1
* Corresponding author. Fax: 46 46 222 8209.
0008-6215/98/$19.00 # 1998 Elsevier Science Ltd. All rights reserved
PII S0008-6215(98)00021-4

244
H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
ꢁ
ꢁ
ꢁ
.
Scheme 1. (a) BF3 OEt2, CH2Cl2, ꢀ22 C, 45 min; (b) MeONa, MeOH, ꢀ22 C, 6 h; (c) CF3SO3Ag, CH2Cl2, ꢀ22 C, 10 h; (d)
Cs2CO3, DMF, ꢀ22 ꢁC, 10 h.
(2.5 equiv), using boron tri¯uoride etherate as
promoter [6], furnishing the thiogalabioside 2
(66%). De-O-acetylation of 2 with methanolic
sodium methoxide gave the divalent glycoside 3
(97%).
(90%). De-O-acetylation of 7 gave the divalent
glycoside 8 (93%).
The thioglycosides 10 and 14 (Scheme 2) were
designed for use as negative controls of multi-
valency in future biological assays, since each
compound carries only one galabiosyl residue.
Glycosylation of benzylthiol with the trichloro-
acetimidate 1 (1.7 equiv.), using boron tri¯uoride
etherate as promoter [6], gave 9 (88%), and de-O-
acetylation yielded the thiogalabioside 10 (93%).
Boron tri¯uoride etherate-induced glycosylation
[10] of 1,3-dimercaptomethylbenzene with the per-
O-acetylated lactose 11 (1.1 equiv) gave the mono-
glycosylated compound 12 (79%). Treatment of 12
with 1 (1.2 equiv) as above furnished 13, con-
taminated by 5% of an unknown compound. De-
O-acetylation of the mixture permitted the isola-
tion of pure 14 (57% overall yield from 12).
Glycosylation of 1,3-dihydroxymethylbenzene
with 1 (2.8 equiv) using silver tri¯uoromethane-
sulfonate [7] as promoter, gave 4 (58%), con-
taminated with 3±5% of an unknown compound.
De-O-acetylation as above gave the divalent gly-
coside 5 (87%) as a pure compound. Attempted
glycosylation with boron tri¯uoride etherate as
promoter was unsuccessful, leading mainly to a
monoglycosylated product where the benzylic
hydroxymethyl group was acetylated. Similarly,
when the benzoyl-protected trichloroacetimidate
analog of 1 was used instead of 1, the product was
monoglycosylated (the benzylic hydroxymethyl
group was unprotected). Addition of a second
portion of the imidate did not lead to a second
glycosylation.
The E. coli proteins PapGJ96 and PapGAD110 use
galabiose and globotriose, respectively, for optimal
recognition [11,12], and we therefore synthesized
the globotriosides 23 and 25 (Scheme 3). The
known [11] 2-(trimethylsilyl)ethyl (TMSEt) globo-
trioside 15 was O-acetylated with acetic anhydride
Alkylation of 1,3-dimercaptomethylbenzene with
the known [8] 2-bromoethyl galabioside 6 (2.4 equiv)
in the presence of cesium carbonate [9] furnished 7

H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
245
in pyridine to yield the protected glycoside 16
(96%). Removal of the TMSEt group of 16 with
tri¯uoroacetic acid in dichloromethane [13] gave
the hemiacetal 17 [14] (90%), and 17 was trans-
formed into the trichloroacetimidate 18 [14] (69%, ꢀ/
ꢁ 10:1) by treatment [6] with trichloroacetonitrile and
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Treat-
ment of 18 with 2-bromoethanol and boron tri-
¯uoride etherate [6] gave the known [15] 2-
bromoethyl globotrioside 19 (58%). The bromine
atom of 19 was substituted by an azido group,
using sodium azide in N,N-dimethylformamide, to
furnish the azido compound 20 (93%). De-O-acet-
ylation of 20 with methanolic sodium methoxide,
followed by catalytic hydrogenation of the azido
group, gave the primary amine 21 (72%).
microbes and microbial proteins to natural glyco-
lipids of the globo series, will be reported in due
course.
3. Experimental
General methods.ÐSee previous paper [2]. Com-
pounds 1 [5], 6 [16], 11 [17], 15 [11], 22 [5], and 24
[5] were synthesized as reported in the literature.
The known [5] carboxylic acid 22 was treated
with N-hydroxysuccinimide to give the corre-
sponding crude NHS-ester, which was used for N-
acylation of the amine 21 to furnish the globo-
triosyl amide 23 (69%). Similarly, the NHS-ester of
the known [5] bis-carboxylic acid 24 acylated the
amine 21 to yield the divalent glycoside 25 (69%).
The inhibitory eciencies of compounds 3, 5, 8,
10, 14, 21, 23, and 25 against the adhesion of
Scheme 3. (a) Ac2O, pyridine, ꢀ22 ꢁC, 12 h; (b) CF3COOH,
CH2Cl2, ꢀ22 ꢁC, 80 min; (c) Cl3CCN, DBU, CH2Cl2, 0 ꢁC, 1 h;
ꢁ
.
(d) HOCH2CH2Br, BF3 OEt2, CH2Cl2, 22 C, 40 min; (e)
NaN3, DMF, 15-crown-5, 75 ꢁC, 24 h; (f) MeONa, MeOH;
i
iiH2, Pd-C, EtOH, HCl, H2O, ꢀ22 ꢁC, 2 h; (g) i22, NHS, EDC,
ꢁ
ii
.
Scheme 2. (a) BF3 OEt2, CH2Cl2, ꢀ22 C, 1 h; (b) MeONa,
DMF, ꢀ22 ꢁC, 18 h, chromatography; 21, DMF, pyridine,
MeOH, ꢀ22 ꢁC.
65 ꢁC, 16 h.

246
H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
1
Acetylated Bis-thiogalabioside 2.ÐTo a solution
of 1 (115 mg, 0.147 mmol) and 1,2-dimercapto-
compound. H NMR (CDCl3): ꢂ 7.21±7.35 (m, 4
H, Ph), 5.59 (bd, 2 H, J 3.3 Hz, H-40), 5.42 (dd, 2
H, J 3.3, 11.1 Hz, H-30), 5.26 (dd, 2 H, J 7.8,
10.7 Hz, H-2), 5.22 (dd, 2 H, J 3.5, 11.1 Hz, H-20),
5.03 (d, 2 H, J 3.6 Hz, H-10), 4.93 (d, 2 H, J
12.4 Hz, PhCH2O), 4.83 (dd, 2 H, J 2.8, 10.8 Hz,
H-3), 4.64 (d, 2 H, J 12.4 Hz, PhCH2O), 4.56 (m, 2
H, H-50), 4.55 (d, 2 H, J 7.8 Hz, H-1), 4.49 (dd, 2
H, J 6.7, 11.2 Hz, H-6a), 4.05±4.24 (m, 8 H, H-
6b,4,60), 3.80 (bt, 2 H, J 6.8 Hz, H-5), 2.14, 2.12,
2.10, 2.08, 2.05, 2.02, and 1.99 (7 s, 42 H, Ac); 13C
NMR (CDCl3): ꢂ 171.1, 171.0, 170.9, 170.86, 170.6,
170.2, 169.5, 137.5, 129.1, 127.8, 127.4, 100.2, 99.8,
77.5, 73.1, 72.4, 70.8, 69.2, 69.0, 68.3, 67.8, 67.5,
62.4, 61.0, 21.3, 21.2, 21.14, 21.1, 21.06; HRMS
calcd for C60H78O36Na (M+Na): 1397.4170,
found: 1397.4188.
methylbenzene (10 mg, 0.059 mmol) in CH2Cl2
.
(0.037 mL,
(2 mL)
was
added
BF3 OEt2
0.294 mmol) under N2. After 45 min at ꢀ22 ꢁC, sat
aq NaHCO3 (2 mL) and CH2Cl2 (15 mL) were
added. The organic layer was dried and con-
centrated, and the residue was chromatographed
(2:1 EtOAc±heptane) to give 2 (54 mg, 66%); [ꢀ]21
d
+5ꢁ (c 0.5, CHCl3); 1H NMR (CDCl3): ꢂ 7.21±7.33
(m, 4 H, Ph), 5.57 (bd, 2 H, J 2.3 Hz, H-40), 5.38
(dd, 2 H, J 3.4, 11.0 Hz, H-30), 5.22 (t, 2 H, J
10.2 Hz, H-2), 5.20 (dd, 2 H, J 3.6, 11.0 Hz, H-20),
5.01 (d, 2 H, J 3.7 Hz, H-10), 4.80 (dd, 2 H, J 2.7,
10.4 Hz, H-3), 4.49 (bt, 2 H, J 7.4 Hz, H-50), 4.45
(dd, 2 H, J 4.4, 11.3 Hz, H-6), 4.30 (d, 2 H, J
9.9 Hz, H-1), 3.74 (bt, 2 H, J 6.4 Hz, H-5), 2.13,
2.12, 2.06, 2.03, 1.99, and 1.98 (6 s, 42 H, Ac); 13C
NMR (CDCl3): ꢂ 171.06, 171.0, 170.92, 170.88,
170.6, 170.2, 169.6, 135.8, 131.5, 128.1, 99.6, 83.0,
77.7, 76.2, 74.2, 69.0, 68.3, 67.8, 67.6, 67.56, 62.8,
61.0, 31.6, 21.3, 21.26, 21.2, 21.1; HRMS calcd for
C60H78O34S2Na (M+Na): 1429.3713, found:
1429.3690.
Bis-galabioside 5.ÐCompound
4
(5.5 mg,
0.004 mmol) was dissolved in dry MeOH (2 mL)
and a catalytic amount of MeONa was added.
After 12 h, the mixture was neutralized with Duo-
lite C436 (H+) resin, and concentrated. The resi-
due was chromatographed (Varian Mega Bond
Elut C18; 1:0!9:1!8:2!7:3!6:4 H2O±MeOH,
5 mL each). The puri®ed product was chromato-
graphed (8:5:1 CH2Cl2±MeOH±H2O) to give 5
Bis-thiogalabioside 3.ÐCompound 2 was dis-
solved in dry MeOH and a catalytic amount of
MeONa was added. After 6 h, the mixture was
neutralized with Duolite C436 (H+) resin, and
concentrated. The residue was chromatographed
(5:5:1 CH2Cl2±MeOH±H2O) to give 3 (22 mg,
ꢁ
1
(2.7 mg, 87%); [ꢀ]22 +56 (c 0.4, H2O); H NMR
d
(D2O): ꢂ 7.34±7.46 (m, 4 H, Ph), 4.88 (d, 2 H, J
11.3 Hz, PhCH2O), 4.87 (d, 2 H, J 4.0 Hz, H-10),
4.7 (2 H, PhCH2O, disturbed by the HDO signal),
4.45 (d, 2 H, J 7.7 Hz, H-1), 4.28 (bt, 2 H, J 6.4 Hz,
H-50), 3.94 (m, 4 H, H-4,40), 3.57±3.86 (m, 16 H,
including H-3,20,30,60), 3.49 (dd, 2 H, J 7.8, 10.0 Hz,
H-2); 13C NMR (CDCl3): ꢂ 137.5, 129.2, 129.0,
102.3, 100.6, 77.5, 75.5, 72.8, 71.6, 71.3, 71.2, 69.6,
69.4, 69.1, 60.9, 60.5; HRMS calcd for
ꢁ
1
97%); [ꢀ]22
32 (c 1.8, H2O); H NMR (D2O): ꢂ
d
7.17±7.32 (m, 4 H, Ph), 4.82 (d, 2 H, J 3.9 Hz, H-
10), 4.22 (d, 2 H, J 9.2 Hz, H-1), 4.21 (bt, 2 H, J
6.1 Hz, H-50), 4.06 (d, 2 H, J 13.5 Hz, PhCH2S),
4.01 (d, 2 H, J 13.5 Hz, PhCH2S), 3.92 (d, 2 H, J
2.6 Hz, H-4), 3.89 (d, 2 H, J 2.6 Hz, H-40), 3.78 (dd,
2 H, J 3.2, 10.5 Hz, H-30), 3.40±3.76 (m, 16 H, H-
2,3,5,6,20,60); 13C NMR (D2O): ꢂ 136.4, 131.3,
128.3, 100.7, 85.1, 79.1, 77.8, 74.1, 71.1, 70.0, 69.5,
69.3, 69.1, 60.8, 60.3, 31.5; HRMS calcd for
C32H51O20S2 (M+H): 819.2415, found: 819.2405.
Acetylated Bis-galabioside 4.ÐTo a solution of
1,3-dihydroxymethylbenzene (5 mg, 0.036 mmol), 1
(76 mg, 0.097 mmol) and CF3SO3Ag (25 mg,
0.097 mmol) was added dry CH2Cl2 (2 mL). The
mixture was stirred under N2 at ꢀ22 ꢁC for 10 h.
Additional CF3SO3Ag (25 mg, 0.097 mmol) and 1
(15 mg, 0.02 mmol) were added. After 12 h, the
mixture was ®ltered through Celite and con-
centrated. The residue was chromatographed
(2:1!3:1!5:1 EtOAc±heptane) to give 4 (29 mg,
58%), contaminated by 3±5% of a co-eluting
C32H50O22Na
809.2676.
(M+Na):
809.2691,
found:
Acetylated Bis-galabioside 7.ÐTo a mixture of
compound 6 (209.5 mg, 0.28 mmol), 1,3-mercapto-
methylbenzene (0.018 mL, 0.118 mmol), and dry
DMF (3 mL), was added Cs2CO3 (103 mg,
0.316 mmol). The mixture was stirred at ꢀ22 ꢁC for
10 h under N2, and H2O (10 mL) and CH2Cl2
(30 mL) were added. The organic layer was dried
and concentrated and the residue was chromato-
graphed (1:1 EtOAc±heptane) to give 7 (158 mg,
90%); [ꢀ]21 +41ꢁ (c 1.1, CHCl3); 1H NMR
d
(CDCl3): ꢂ 7.16±7.30 (m, 4 H, Ph), 5.55 (bd, 2 H, J
2.3 Hz, H-40), 5.37 (dd, 2 H, J 3.3, 11.0 Hz, H-30),
5.19 (dd, 2 H, J 3.8, 11.0 Hz, H-20), 5.17 (dd, 2 H, J
8.0, 11.1 Hz, H-2), 4.99 (d, 2 H, J 3.6 Hz, H-10),

H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
247
4.80 (dd, 2 H, J 2.6, 10.8 Hz, H-3), 4.51 (bt, 2 H, J
7.0 Hz, H-50), 4.47 (d, 2 H, J 7.8 Hz, H-1), 4.43 (dd,
2 H, J 6.8, 11.3 Hz, H-6a), 4.04±4.20 (m, 8 H,
2.11, 2.08, 2.05, 2.02, and 2.00 (7 s, 21 H, Ac); 13C
NMR (CDCl3): ꢂ 171.03, 171.0, 170.9, 170.6, 170.2,
169.7, 137.5, 129.5, 129.0, 127.7, 99.5, 82.8, 77.7,
76.3, 74.3, 68.9, 68.3, 67.8, 67.6, 67.5, 62.8, 61.0,
34.0, 21.4, 21.2, 21.16, 21.1, 21.07; HRMS calcd
for C33H42O17SNa (M+Na): 765.2040, found:
765.2040.
including H-4,6b,60), 3.94±4.04 (m,
2
H,
OCH2CH2S), 3.78 (bt, 2 H, J 6.4 Hz, H-5), 3.73 (s,
4 H, PhCH2), 3.56-3.66 (m, 2 H, OCH2CH2S),
2.57±2.72 (m, 4 H, OCH2CH2S), 2.12, 2.09, 2.07,
2.04, 2.02, and 1.98 (6 s, 42 H, Ac); 13C NMR
(CDCl3): ꢂ 171.1, 171.0, 170.9, 170.85, 170.6, 170.2,
169.6, 139.1, 129.7, 129.1, 128.1, 101.6, 99.8, 73.1,
72.4, 69.6, 69.0, 68.3, 67.8, 67.5, 62.4, 60.9, 36.9,
31.2, 21.4, 21.2, 21.14, 21.09, 21.06; HRMS calcd
for C64H86O36S2Na (M+Na): 1517.4238, found:
1517.4243.
Benzyl (a-d-galactopyranosyl)-(1!4)-1-thio-b-
d-galactopyranoside (10).ÐCompound 9 (45 mg,
0.06 mmol) was dissolved in dry MeOH (3 mL) and
a catalytic amount of 0.5 M MeONa was added.
After 10 h, the mixture was neutralized with Duo-
lite C436 (H+) resin and concentrated. The residue
was chromatographed (10:5:1 CH2Cl2±MeOH±
Bis-galabioside 8.ÐCompound
7
(100 mg,
H2O) to give 10 (25 mg, 93%); [ꢀ]22 +7ꢁ (c 0.3,
d
1
0.067 mmol) was dissolved in dry MeOH and a
catalytic amount of 0.5 M MeONa was added.
After 10 h, the mixture was neutralized with Duo-
lite C436 (H+) resin and concentrated. The residue
was chromatographed (10:5:1 CH2Cl2±MeOH±
H2O); H NMR (D2O): ꢂ 7.17±7.35 (m, 5 H, Ph),
4.80 (d, 1 H, J 3.4 Hz, H-10), 04.19 (d, 1 H, J 7.7 Hz,
H-1), 4.14±4.25 (m, 1 H, H-5 ); 13C NMR (D2O): ꢂ
138.4, 129.1, 127.7, 100.7, 84.9, 79.2, 77.9, 74.1,
71.1, 70.06, 69.5, 69.3, 69.1, 60.8, 60.4, 34.2;
HRMS calcd for C19H28O10SNa (M+Na):
471.1301, found: 471.1316.
H2O) to give 8 (56 mg, 93%); [ꢀ]21 +72ꢁ (c 0.7,
d
1
D2O); H NMR (D2O): ꢂ 7.15±7.29 (m, 4 H, Ph),
4.84 (d, 2 H, J 3.9 Hz, H-10), 4.26 (d, 2 H, J 7.8 Hz,
H-1), 4.23 (bt, 2 H, J 6.7 Hz, H-50), 3.88±3.92 (m, 4
H, H-4,40), 3.52±3.87 (m, 24 H, including H-
3,5,20,30 and OCH2CH2S), 3.41 (dd, 2 H, J 7.7,
10.2 Hz, H-2), 2.60 (dt, 4 H, J 0.9, 6.8 Hz,
OCH2CH2S); 13C NMR (D2O): ꢂ 139.4, 129.8,
129.5, 128.2, 103.4, 100.6, 77.4, 75.3, 72.7, 71.23,
71.15, 69.5, 69.3, 69.2, 69.1, 60.9, 60.3, 35.5, 30.6;
HRMS calcd for C36H58O22S2Na (M+Na):
929.2759, found: 929.2763.
(3-Mercaptomethyl)benzyl (2,3,4,6-tetra-O-acetyl-
-b-d-galactopyranosyl)-(1!4)-2,3,6-tri-O-acetyl-
1-thio-b-d-glucopyranoside (12).ÐTo a mixture of
octaacetyl-ꢁ-lactose 11 [17] (87 mg, 0.128 mmol),
1,3-mercaptomethylbenzene (0.018 mL, 0.118 mmol),
.
and CH2Cl2 (2 mL), was added BF3 OEt2
(0.074 mL, 0.587 mmol) under N2. After 1 h at
ꢀ22 ꢁC, sat aq NaHCO3 (2 mL) and CH2Cl2
(10 mL) were added. The organic layer was dried
and concentrated, and the residue was chromato-
graphed (1:1 EtOAc±heptane) to give 12 (73 mg,
Benzyl (2,3,4,6-tetra-O-acetyl-a-d-galactopyrano-
syl)-(1!4)-2,3,6-tri-O-acetyl-1-thio-b-d-galactopyr-
anoside (9).ÐTo a mixture of 1 (50 mg, 0.064
mmol), benzylthiol (0.007 mL, 0.06 mmol), and
79%); [ꢀ]21
43ꢁ (c 0.8, CHCl3); 1H NMR
d
(CDCl3): ꢂ 7.14±7.30 (m, 4 H, Ph), 5.34 (d, 1 H, J
3.2 Hz, H-40), 5.13 (t, 1 H, J 9.9 Hz, H-3), 5.09 (dd,
1 H, J 7.9, 10.4 Hz, H-20), 4.97 (t, 1 H, J 9.9 Hz, H-
2), 4.95 (dd, 1 H, J 3.4, 10.4 Hz, H-30), 4.49 (m, 1
H, H-6a), 4.47 (d, 1 H, J 7.7 Hz, H-10), 4.26 (d, 1
H, J 10.1 Hz, H-1), 4.03±4.16 (m, 3 H, H-6b,60),
3.89 (d, 1 H, J 12.8 Hz, PhCH2SC), 3.86 (bt, 1 H, J
6.7 Hz, H-50), 3.74±3.82 (m, 2 H, H-4 and
PhCH2SC), 3.73 (d, 2 H, J 7.6 Hz, PhCH2SH), 3.54
(ddd, 1 H, J 1.6, 5.4, 9.9 Hz, H-5), 2.15, 2.14, 2.05,
2.03, 2.01, and 1.96 (6 s, 21 H, Ac), 1.78 (t, 1 H, J
7.6 Hz, SH); 13C NMR (CDCl3): ꢂ 170.8, 170.6,
170.5, 170.11, 170.10, 169.5, 142.0, 137.7, 129.4,
129.2, 128.2, 127.6, 101.5, 82.0, 77.0, 76.7, 74.2,
71.4, 71.1, 70.6, 69.5, 67.0, 62.7, 61.2, 34.1, 29.2,
21.4, 21.2, 21.14, 21.1, 20.9; HRMS calcd for
C34H44O17S2Na (M+Na): 811.1918, found:
811.1918.
.
CH2Cl2 (2 mL), was added BF3 OEt2 (0.008 mL,
0.064 mmol) under N2. After 1 h at ꢀ22 ꢁC, sat aq
NaHCO3 (10 mL) and CH2Cl2 (20 mL) were
added. The organic layer was dried and con-
centrated, and the residue was chromatographed
(2:1 EtOAc±heptane) to give 9 (38 mg, 88%); [ꢀ]21
d
ꢁ
1
+36 (c 1.4, CHCl3); H NMR (CDCl3): ꢂ 7.30±
7.37 (m, 5 H, Ph), 5.58 (bd, 1 H, J 3.3 Hz, H-40),
5.40 (dd, 1 H, J 3.3, 11.0 Hz, H-30), 5.29 (t, 1 H, J
10.1 Hz, H-2), 5.22 (dd, 1 H, J 3.7, 11.1 Hz, H-20),
5.03 (d, 1 H, J 3.7 Hz, H-10), 4.82 (dd, 1 H, J 2.7,
10.3 Hz, H-3), 4.50 (bt, 1 H, J 6.5 Hz, H-50), 4.45
(dd, 1 H, J 7.1, 11.4 Hz, H-6a), 4.31 (d, 1 H, J
9.9 Hz, H-1), 4.06±4.20 (m, 4 H, H-4,6b,60), 3.98 (d,
1 H, J 12.9 Hz, PhCH2), 3.87 (d, 1 H, J 12.9 Hz,
PhCH2), 3.74 (bt, 1 H, J 6.5 Hz, H-5), 2.14, 2.12,

248
Acetylated
(13).ÐTo a mixture of compound 12 (40 mg,
0.051 mmol), compound 1 (47.5 mg, 0.061 mmol),
.
and CH2Cl2 (2 mL), was added BF3 OEt2
H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
thiolactosyl-thiogalabiosyl
dimer
71.1, 70.1, 69.5, 69.3, 69.1, 68.9, 61.4, 60.8, 60.6,
60.5, 33.7, 33.6; HRMS calcd for C32H50O20S2Na
(M+Na): 841.2235, found: 841.2260.
2-(Trimethylsilyl)ethyl (2,3,4,6-tetra-O-acetyl-
a-d-galactopyranosyl)-(1!4)-(2,3,6-tri-O-acetyl-b-
d - galactopyranosyl) - (1!4) - 2,3,6-tri-O-acetyl-
b-d-glucopyranoside (16).ÐThe TMSEt globotrio-
side 15 [11] (34 mg, 0.056 mmol) was dissolved in
pyridine (6 mL) and Ac2O (6 mL). The mixture was
stirred at ꢀ22 ꢁC for 12 h, toluene (30 mL) was
added, and the mixture was concentrated. The resi-
due was chromatographed (1:1 EtOAc±heptane) to
(0.015 mL, 0.12 mmol) under N2. After 1 h at
ꢀ22 ꢁC, sat aq NaHCO3 (2 mL) and CH2Cl2
(15 mL) were added. The organic layer was dried
and concentrated, and the residue was chromato-
graphed (2:1 EtOAc±heptane) to give 13 con-
taminated with 5% of an unknown compound
(54 mg). An analytical sample of 13 was obtained
by rechromatography of crude 13; [ꢀ]21
14ꢁ (c
d
1
0.3, CHCl3); H NMR (CDCl3): ꢂ 7.15±7.30 (m, 4
H, Ph), 5.58 (bd, 1 H, J 3.2 Hz, H-40gala), 5.40 (dd, 1
H, J 3.3, 11.0 Hz, H-30gala), 5.36 (bd, 1 H, J 3.3 Hz,
H-40lac), 5.30 (t, 1 H, J 10.1 Hz, H-2gala), 5.22 (dd,
1 H, J 3.6, 11.1 Hz, H-20gala), 5.16 (t, 1 H, J 9.2 Hz,
H-3lac), 5.11 (dd, 1 H, J 7.9, 10.4 Hz, H-20lac), 5.03
(d, 1 H, J 3.5 Hz, H-10gala), 4.98 (t, 1 H, J 9.9 Hz,
H-2lac), 4.97 (dd, 1 H, J 3.4, 10.4 Hz, H-30lac), 4.85
(dd, 1 H, J 2.7, 10.4 Hz, H-3gala), 4.51 (m, 1 H, H-
50gala), 4.49 (d, 1 H, J 7.9 Hz, H-10lac), 4.45 (dd, 1
H, J 7.0, 11.4 Hz, H-6alac), 4.35 (d, 1 H, J 10.1 Hz,
H-1lac), 4.34 (d, 1 H, J 9.8 Hz, H-1gala), 4.05±4.20
(m, 7 H, including H-4gala, H-6blac), 3.75±4.00 (m,
7 H, including H-4lac), 3.58 (m, 1 H, H-5lac), 2.165,
2.160, 2.15, 2.13, 2.12, 2.09, 2.07, 2.065, 2.05,
2.045, 2.035, 2.030, 2.00, and 1.98 (14 s, 42 H, Ac);
13C NMR (CDCl3): ꢂ 171.0, 170.9, 170.8, 170.6,
170.2, 169.5, 138.0, 137.7, 130.1, 129.2, 128.7,
128.4, 101.6, 99.6, 82.9, 82.4, 76.7, 76.3, 74.2, 71.4,
71.1, 70.6, 69.5, 68.9, 68.3, 67.7, 67.6, 67.5, 67.0,
62.6, 61.2, 61.0, 34.1, 33.7, 21.4, 21.2, 21.1, 20.9;
HRMS calcd for C60H78O34S2Na (M+Na):
1429.3714, found: 1429.3721.
give 16 (55 mg, 96%); [ꢀ]21 +40ꢁ (c 2.2, CHCl3);
d
1H NMR (CDCl3): ꢂ 5.60 (bd, 1 H, J 2.2 Hz, H-400),
5.40 (dd, 1 H, J 3.3, 11.0 Hz, H-300), 5.21 (t, 1 H, J
9.2 Hz, H-3), 5.19 (dd, 1 H, J 3.6, 11.1 Hz, H-200),
5.11 (dd, 1 H, J 7.7, 10.8 Hz, H-20), 4.99 (d, 1 H, J
3.6 Hz, H-100), 4.88 (dd, 1 H, J 7.9, 9.4 Hz, H-2),
4.74 (dd, 1 H, J 2.6, 10.8, H-30), 4.52 (d, 1 H, J
7.7 Hz, H-10), 4.50 (d, 1 H, J 7.9 Hz, H-1), 4.41±4.53
(m, 3 H, H-500,6a,60a), 4.07±4.21 (m, 4 H, H-6b,60b,
600), 4.02 (d, 1 H, J 2.1 Hz, H-40), 3.95 (dt, 1 H, J 5.8,
10.0 Hz, CH2CH2O), 3.73±3.85 (m, 2 H, H-4,50),
3.61±3,67 (m, 1 H, H-5), 3.57 (dt, 1 H, J 6.9, 9.9 Hz,
CH2CH2O), 2.14, 2.12, 2.09, 2.08, 2.075, 2.070,
2.065, 2.06, 2.04, and 1.99 (10 s, 30 H, Ac), 0.79±
1.01 (m, 2 H, CH2SiMe3), 0.01 (s, 9 H, SiMe3); 13
C
NMR (CDCl3): ꢂ 171.1, 170.9, 170.5, 170.2, 170.0,
169.3, 101.6, 100.4, 100.1, 77.4, 73.8, 73.3, 72.9,
72.3, 72.2, 69.4, 69.3, 68.3, 67.9, 67.6, 67.5, 62.8,
61.7, 60.7, 21.4, 21.3, 21.2, 21.15, 21.1, 21.0, 20.9,
18.3, 1.0; HRMS calcd for C43H64O26SiNa
(M+Na): 1047.3353, found: 1047.3344.
(2,3,4,6 - Tetra - O- acetyl - a-d-galactopyranosyl) -
(1!4) - (2,3,6-tri-O-acetyl-b-d-galactopyranosyl)-
Thiolactosyl-thiogalabiosyl dimer 14.ÐCrude 13
(42 mg) was dissolved in MeOH and MeONa
(0.5 M) was added. After 6 h, the mixture was
neutralized with Duolite C436 (H+) resin and
concentrated. The residue was chromatographed
(Varian Mega Bond Elut Column C18;
9:1!8:2!7:3!6:4!5:5 H2O±MeOH, 5 mL each)
(1!4)-2,3,6-tri-O-acetyl-b-d-glucopyranose
(17)
[14].ÐTo a solution of compound 16 (250 mg,
0.24 mmol) in CH2Cl2 (1.2 mL) was added
CF3COOH (2.4 mL, 0.031 mmol), and the mixture
was stirred at ꢀ22 ꢁC under N2. After 80 min, n-
propyl acetate (10 mL) and toluene (10 mL) were
added, and the mixture was concentrated and co-
concentrated twice with toluene. The residue was
chromatographed (3:1 EtOAc±heptane) to give
the hemiacetal 17 (204 mg, 90%); HRMS calcd
for C38H52O26Na (M+Na): 947.2645, found:
947.2652.
to give pure 14 (18.4 mg, 57% overall yield from
12); [ꢀ]21
77ꢁ (c 1.3, H2O); H NMR (D2O):
1
d
ꢂ0 7.16±7.30 (m, 4 H, Ph), 4.80 (d, 10H, J 3.9 Hz, H-
1 gala), 4.31 (d, 1 H, J 7.8 Hz, H-1 lac), 4.21 (bt, 1
H, J 6.6 Hz, H-50gala), 4.15 (d, 1 H, J 9.9 Hz, H-
1lac), 4.12 (bd, 1 H, J 9.6 Hz, H-1gala), 3.86±3.94
(m, 4 H), 3.30±3.84 (m, 22 H), 3.27 (dd, 1 H, J 9.0,
9.9 Hz, H-2lac); 13C NMR (D2O): ꢂ 138.7, 138.6,
130.1, 129.4, 128.4, 103.2, 100.7, 84.4, 83.8, 79.2,
78.9, 78.5, 77.9, 76.1, 75.7, 74.1, 72.9, 72.3, 71.3,
(2,3,4,6-Tetra-O-acetyl-a-d-galactopyranosyl)-
(1!4)-(2,3,6-tri-O-acetyl-b-d-galactopyranosyl)-
(1!4)-2,3,6-tri-O-acetyl-a-d-glucopyranosyl tri-
chloroacetimidate (18) [14].ÐCompound 17
(204 mg, 0.22 mmol) was dissolved in a mixture of

H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
249
dry CH2Cl2 (3.4 mL) and dry Cl3CCN (0.85 mL,
8.4 mmol). The mixture was cooled to 0 ꢁC under
N2, and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU;
0.051 mL, 0.34 mmol) was added. The mixture was
stirred at 0 ꢁC for 1 h, then washed with cold H2O,
dried, and concentrated. The residue was chroma-
tographed (2:1 EtOAc±heptane) to give 18 (163 mg,
3.73±3.86 (m, 3 H, H-4,50 and OCH2CH2Br), 3.64
(ddd, 1 H, J 1.9, 5.0, 9.8 Hz, H-50), 3.39±3.49 (m, 2
H, CH2Br), 2.13, 2.12, 2.08, 2.065, 2.060, 2.055,
2.05, 2.045, and 1.98 (9 s, 30 H, Ac); 13C NMR
(CDCl3): ꢂ 171.1, 170.9, 170.86, 170.8, 170.5, 170.2,
170.0, 169.95, 169.3, 101.5, 101.2, 100.1, 77.3, 76.8,
73.3, 73.2, 73.1, 72.3, 71.9, 70.2, 69.4, 69.3, 68.3,
67.6, 67.5, 62.5, 61.8, 60.7, 30.2, 21.34, 21.26, 21.2,
21.12, 21.1, 21.05, 21.0, 20.9; HRMS calcd for
C40H55O26BrNa (M+Na): 1053.2063, found:
1053.2089.
1
69%) as an ꢀ/ꢁ mixture (10:1). H NMR for 18ꢀ
(CDCl3): ꢂ 8.66 (s, 1 H, =NH), 6.49 (d, 1 H, J
3.8 Hz, H-1), 5.60 (bs, 1 H, H-400), 5.57 (t, 1 H, J
9.5 Hz, H-3), 5.41 (dd, 1 H, J 3.4, 11.1 Hz, H-300),
5.19 (dd, 1 H, J 3.5, 10.9 Hz, H-200), 5.13 (dd, 1 H, J
7.8, 10.8 Hz, H-20), 5.08 (dd, 1 H, J 3.8, 10.2 Hz, H-
2), 5.00 (d, 1 H, J 3.5 Hz, H-100), 4.75 (dd, 1 H, J
2.4, 10.8 Hz, H-30), 4.55 (d, 1 H, J 7.8 Hz, H-10),
4.42±4.55 (m, 3 H, H-500,60a,6a), 4.07±4.22 (m, 5 H,
H-600,60b,5,6b), 4.03 (d, 1 H, J 2.2 Hz, H-40), 3.88 (t,
1 H, J 9.4 Hz, H-4), 3.78 (t, 1 H, J 6.7 Hz, H-50),
2.14, 2.11, 2.10, 2.09, 2.08, 2.07, 2.06, 2.05, 2.02,
and 1.99 (10 s, 30 H, Ac); 13C NMR (CDCl3): ꢂ
171.1, 170.9, 170.7, 170.54, 170.5, 169.9, 169.7,
169.2, 161.5, 101.7, 100.0, 93.4, 76.4, 73.4, 72.3,
71.4, 70.4, 70.2, 69.5, 69.4, 68.3, 67.6, 67.5, 62.1,
61.8, 60.7, 21.4, 21.2, 21.15, 21.1, 21.0, 20.9;
HRMS calcd for C40H52O26Cl3NNa (M+Na):
2-Azidoethyl (2,3,4,6-tetra-O-acetyl-a-d-galacto-
pyranosyl)-(1!4)-(2,3,6-tri-O-acetyl-b-d-galacto-
pyranosyl)-(1!4)-2,3,6-tri-O-acetyl-b-d-glucopyr-
anoside (20).ÐTo a solution of compound 19
(75 mg, 0.073 mmol) in DMF (3 mL) were added
NaN3 (15 mg, 0.23 mmol) and 15-crown-5
(0.015 mL, 0.073 mmol), and the mixture was stir-
red at 75 ꢁC. After 24 h, the mixture was cooled to
ꢀ22 ꢁC and H2O (5 mL) and Et2O (20 mL) were
added. The organic layer was isolated, dried, and
concentrated. The residue was chromatographed
(2:1 EtOAc±heptane) to give 20 (67 mg, 93%);
[ꢀ]22 +35ꢁ (c 1.2 , CHCl3); H NMR (CDCl3): ꢂ
1
d
5.58 (bd, 1 H, J 3.4 Hz, H-400), 5.39 (dd, 1 H, J 3.4,
1090.1741, found: 1090.1757. H NMR for 18ꢁ
11.0 Hz, H-300), 5.21 (t, 1 H, J 9.1 Hz, H-3), 5.18
(dd, 1 H, J 3.6, 11.0 Hz, H-200), 5.11 (dd, 1 H, J 7.7,
10.8 Hz, H-20), 4.99 (d, 1 H, J 3.6 Hz, H-100), 4.92
(dd, 1 H, J 7.8, 9.3 Hz, H-2), 4.74 (dd, 1 H, J 2.5,
10.8 Hz, H-30), 4.57 (d, 1 H, J 7.8 Hz, H-1), 4.53 (d,
1 H, J 7.6 Hz, H-10), 4.45±4.54 (m, 2 H, H-500,6a),
4.43 (dd, 1 H, J 6.3, 11.1 Hz, H-60a), 4.06±4.20 (m,
4 H, H-600,60b,6b), 4.01 (bd, 1 H, J 2.3 Hz, H-40),
3.98 (ddd, 1 H, J 3.6, 5.0, 8.6 Hz, OCH2CH2N3),
3.82 (t, 1 H, J 9.6 Hz, H-4), 3.76 (dt, 1 H, J 6.9 Hz,
H-50), 3.61±3.73 (m, 2 H, H-5 and OCH2CH2N3),
3.47 (ddd, 1 H, J 3.4, 8.2, 13.3 Hz, CH2CH2N3),
3.27 (ddd, 1 H, J 3.4, 4.8, 13.3 Hz, CH2CH2N3),
2.13, 2.11, 2.08, 2.075, 2.065, 2.06, 2.05, 2.04, and
1.98 (9 s, 30 H, Ac); 13C NMR (CDCl3): ꢂ 171.1,
170.9, 170.86, 170.8, 170.5, 170.2, 170.1, 170.0,
169.3, 101.5, 100.8, 100.1, 77.3, 76.8, 73.6, 73.2,
73.1, 72.3, 72.0, 69.4, 69.3, 69.1, 68.3, 67.6, 67.5,
62.4, 61.7, 60.7, 50.9, 21.3, 21.25, 21.13, 21.1,
21.04, 21.0, 20.9; HRMS calcd for C40H55O26N3Na
(M+Na): 1016.2971, found: 1016.2980.
1
(CDCl3): ꢂ 5.89 (d, 1 H, J 7.7 Hz, H-1), the
remaining signals were obscured by the signals of
the main component 18ꢀ.
2-Bromoethyl (2,3,4,6-tetra-O-acetyl-a-d-galacto-
pyranosyl)-(1!4)-(2,3,6-tri-O-acetyl-b-d-galacto-
pyranosyl)-(1!4)-2,3,6-tri-O-acetyl-b-d-glucopyr-
anoside (19) [15].ÐA mixture of compound 18
(139 mg, 0.13 mmol), 2-bromoethanol (0.015 mL,
0.ꢁ21 mmol), and CH2Cl2 (3.5 mL) was cooled to
.
0 C, and BF3 OEt2 (0.016 mL, 0.13 mmol) was
added. The mixture was stirred at ꢀ22 ꢁC under
N2. After 40 min, sat aq NaHCO3 (2 mL) and
CH2Cl2 (10 mL) were added. The organic layer was
isolated, dried, and concentrated. The residue was
chromatographed (3:1 EtOAc±heptane) to give 19
(78 mg, 58%); [ꢀ]22d +45ꢁ (c 1.0, CHCl3), lit +45ꢁ
1
[15]; H NMR (CDCl3): ꢂ 5.59 (bd, 1 H, J 3.2 Hz,
H-400), 5.39 (dd, 1 H, J 3.3, 11.0 Hz, H-300), 5.21 (t,
1 H, J 9.1 Hz, H-3), 5.18 (dd, 1 H, J 3.6, 11.0 Hz,
H-200), 5.11 (dd, 1 H, J 7.8, 10.9 Hz, H-20), 4.99 (d,
1 H, J 3.6 Hz, H-100), 4.92 (dd, 1 H, J 7.9, 9.4 Hz,
H-2), 4.74 (dd, 1 H, J 2.6, 10.8 Hz, H-30), 4.55 (d, 1
H, J 7.9 Hz, H-1), 4.52 (d, 1 H, J 7.8 Hz, H-10),
4.44±4.52 (m, 2 H, H-500,6a), 4.43 (dd, 1 H, J 6.2,
11.1 Hz, H-60a), 4.06±4.20 (m, 5 H, H-60b,6b,600
and OCH2CH2Br), 4.01 (bd, 1 H, J 2.4 Hz, H-40),
2-Aminoethyl
(a-d-galactopyranosyl)-(1!4)-
(b-d-galactopyranosyl)-(1!4)-b-d-glucopyranoside
(21).ÐCompound 20 (65 mg, 0.065 mmol) was dis-
solved in MeOH and a catalytic amount of
MeONa (0.5 M) was added. After 12 h, the mixture
was neutralized with Duolite C436 (H+) resin and

250
H.C. Hansen, G. Magnusson/Carbohydrate Research 307 (1998) 243±251
concentrated. The residue was dissolved in a mix-
ture of EtOH (7 mL) and 0.1 M aq HCl (0.65 mL,
0.065 mmol), and hydrogenated (H2, 10% Pd-C, 1
atm) for 2 h. The mixture was ®ltered through
Celite, passed through a column packed with
Duolite A147 (OH ) resin and concentrated. The
residue was chromatographed (Varian Mega Bond
Elut Column C18; 9:1!8:2!7:3 H2O±MeOH,
!5:5 H2O±MeOH, 6 mL each). The product was
chromatographed on SiO2 (5:5:1 CH2Cl2±MeOH±
H2O!1:1 MeOH±H2O) to give 25 (7.5 mg, 69%);
[ꢀ]23d +48ꢁ (c 0.75, H2O); 1H NMR (D2O): ꢂ 7.18±
7.33 (4 H, Ph), 4.86 (d, 2 H, J 3.8 Hz, H-100), 4.41
and 4.40 (2 d, each 1 H, J 7.7 and 8.0 Hz, H-10,1),
4.26 (bt, 2 H, J 6.3 Hz, H-500), 3.92±3.97 (m, 4 H,
H-400,40), 3.23 (bdd, 2 H, J 8.0, 9.2 Hz, H-20 or H-
2), 2.64 (t, 4 H, J 6.9 Hz, SCH2CH2CO), 2.43 (t, 4
H, J 6.9 Hz, SCH2CH2CO); 13C NMR (D2O): ꢂ
175.0, 139.3, 129.7, 129.6, 128.1, 103.7, 102.6,
100.7, 79.1, 77.8, 75.8, 75.2, 74.7, 73.3, 72.6, 71.3,
71.2, 69.5, 69.4, 69.0, 68.9, 60.9, 60.8, 60.5, 39.8,
35.8, 35.4, 27.0; HRMS calcd for C54H88O34N2
S2Na (M+Na): 1395.4558, found: 1395.4541.
6 mL each) to give 21 (25.8 mg, 72%); [ꢀ]22 +51ꢁ
d
1
(c 1.0, H2O); H NMR (D2O): ꢂ 4.86 (d, 1 H, J
3.9 Hz, H-100), 4.39±4.44 (2 d, 2 H, J 7.8, 8.0 Hz, H-
10,1), 4.26 (bt, 1 H, J 6.6 Hz, H-500), 3.95 (bd, 1 H,
J 3.2 Hz, H-40), 3.94 (bd, 1 H, J 3.3 Hz, H-400),
3.20±3.28 (m, 1 H, H-2), 3.10±3.20 and 2.77±2.90
(multiplets, 2 H, CH2NH2/CH2NH3+); 13C NMR
(D2O): ꢂ 103.7, 102.5, 100.7, 79.1, 77.8, 75.8, 75.2,
74.8, 73.3, 72.6, 71.3, 71.2, 70.4, 69.5, 69.4, 69.0,
60.9, 60.8, 60.4; HRMS calcd for C20H38O16N
(M+H): 548.2191, found: 548.2197.
Acknowledgements
This work was supported by grants from the
Swedish Natural Science Research Council and the
National Institutes of Health (NIH, USA).
2-(5-Phenyl-4-thiapentanoylamido)ethyl (a-d-
galactopyranosyl)-(1!4)-(b-d-galactopyranosyl)-
(1!4)-b-d-glucopyranoside (23).ÐTreatment of
compound 22 with N-hydroxysuccinimide (NHS)
furnished the corresponding NHS-ester, as descri-
bed [5]. The NHS-ester (6 mg, 0.02 mmol) and
compound 21 (10 mg, 0.018 mmol) were dissolved
in 1:1 DMF-pyridine (1 mL), and the mixture was
stirred at 60 ꢁC. After 10 h, the mixture was cooled
to ꢀ22 ꢁC and concentrated, and the residue was
chromatographed (Varian Mega Bond Elut C18;
9:1!8:2!7:3!6:4!5:5!4:6 H2O±MeOH, 5 mL
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each) to give 23 (9 mg, 69%); [ꢀ]d +45ꢁ (c 0.6,
23
1
H2O); H NMR (D2O): ꢂ 7.16±7.29 (m, 5 H, Ph),
4.80 (d, 1 H, J 3.9 Hz, H-100), 4.34 (d, 2 H, J 7.9 Hz,
H-10,1), 4.20 (bt, 1 H, J 6.7 Hz, H-500), 2.59 (t, 2
H, J 6.9 Hz, SCH2CH2CO), 2.38 (t, 2 H, J 6.9 Hz,
SCH2CH2CO); 13C NMR (D2O): ꢂ 175.0, 138.7,
129.3, 129.2, 127.7, 103.6, 102.5, 100.7, 79.0, 77.7,
75.8, 75.1, 74.7, 73.2, 72.5, 71.2, 69.4, 69.2, 68.8,
60.8, 60.7, 60.3, 39.7, 35.7, 35.5, 27.0; HRMS calcd
for C30H48O17NS (M+H): 726.2642, found:
726.2637.
Bis-globotrioside 25.ÐTreatment of compound
24 with N-hydroxysuccinimide (NHS) furnished
the corresponding NHS-ester, as described [5]. A
solution of compound 21 (18 mg, 0.032 mmol) in
freshly distilled DMF (3 mL) was added to a solu-
tion of the crude NHS-ester (4 mg, 0.0079 mmol) in
pyridine (1 mL), and the mixture was stirred at
65 ꢁC. After 16 h, the mixture was concentrated,
and the residue was chromatographed (Varian
Mega Bond Elut C18; 1:0!9:1!8:2!7:3!6:4

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