Synthesis of octyl S-glycosides of tri- to pentasaccharide fragments related to the GPI anchor of Trypanosoma brucei

Article abstract of DOI:10.1016/j.carres.2011.08.001

The three oligosaccharide octyl-S-glycosides Man-α1,6-Man-α1,4- GlcNH₂-α1,S-Octyl (19), Man-α1,6-(Gal-α1,3)Man- α1,4-GlcNH₂-α1,S-Octyl (27) and Man-α1,2-Man- α1,6-(Gal-α1,3)Man-α1,4-GlcNH₂-α1,S-Octyl (37), related to the G

Full text of DOI:10.1016/j.carres.2011.08.001

Carbohydrate Research 346 (2011) 2348–2361
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Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Synthesis of octyl S-glycosides of tri- to pentasaccharide fragments related
to the GPI anchor of Trypanosoma brucei
⇑
Ralf Dettmann, Thomas Ziegler
Institute of Organic Chemistry, University of Tuebingen, Auf der Morgenstelle 18, 72076 Tuebingen, Germany
a r t i c l e i n f o
a b s t r a c t
Article history:
The three oligosaccharide octyl-S-glycosides Man-
a
1,6-Man-
1,2-Man-
1,S-Octyl (37), related to the GPI anchor of Trypanosoma brucei were prepared by a stepwise and
block-wise approach from octyl 2-azido-2-deoxy-3,6-di-O-benzyl-1-thio- -glucopyranoside (8) and
octyl 2-O-benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane-1,3-diyl)-1-thio- -mannopyransoside
(9). Glucosamine derivative 8 was obtained from 1,3,4,6-tetra-O-acetyl-2-azido-2-desoxy-b- -glucopyr-
a
1,4-GlcNH2-
a
1,S-Octyl (19), Man-
a1,6-
Received 22 March 2011
Received in revised form 21 June 2011
Accepted 2 August 2011
(Gal-
a
1,3)Man-
a
1,4-GlcNH2-
a1,S-Octyl (27) and Man-
a
a1,6-(Gal-
a
1,3)Man- 1,4-GlcNH2-
a
a
a
-D
Available online 10 August 2011
a
-D
D
Keywords:
anose (1) in five steps. Mannoside 9 was converted into the corresponding imidate 12 and coupled with
8 to give disaccharide octyl-S-glycoside 13 which was further mannosylated to afford trisaccharide 19
upon deprotection. Likewise, mannoside 9 was galactosylated, converted into the corresponding imidate
and coupled with 8 to give trisaccharide 25. Mannosylation of the latter afforded tetrasaccharide 27 upon
deprotection. Condensation of 25 with disaccharide imidate 35 gave, upon deprotection of the interme-
diates, the corresponding pentasaccharide octyl-S-glycoside 37. Saccharides 19, 27 and 37 are suitable
substrates for studying the enzymatic glycosylation pattern of the GPI anchor of T. brucei.
Ó 2011 Elsevier Ltd. All rights reserved.
GPI anchor
Glycosylation
Glycodesilylation
1. Introduction
(ABCDE, Fig. 1) with modifications at the mannose residues
depending on the respective parasite or source of the GPI-anchor.
The pathogenic organism of the often deadly sleeping sickness
in humans (trypanosomiasis) and in cattle (animal trypanosomia-
sis or Nagana) is the tsetse fly-transmitted protozoan Trypanosoma
brucei. This pathogenic parasite belongs to the class Kinetoplastidae,
suborder Flagellae, and lives in the blood stream of its host causing
strong sleeping disorders. In advanced stages of the disease the
protozoan invade the central nervous system causing severe
sleep-like unconsciousness, coma and death. The parasites are
usually destroyed efficiently by the immune system of the host.
However, some trypanosomes evade the immune system and final-
ly exhaust the host’s defense through antigenic variation. This
variation is achieved by the protozoan by constantly modifying
its antigenic surface structures due to a highly variant surface
glycoprotein (VSG) whose presence on the cell surface can well
exceed 107 copies.1–3
In case of the VSG-GPI anchor of T. brucei the side-chains could
be identified as the VSG-Protein attached to the ethanolamino-
phosphate residue and a branched a-glactosyl-residue of variable
length (F, Fig. 1) attached to the first mannosyl residue of the core.
Formerly, great efforts were taken for the total synthesis of di-
verse GPI-anchors.5,6 Examples of partial syntheses of this struc-
ture were published too.7–12 Previously, we have introduced a
new efficient synthetic strategy for the chemical synthesis of di-
verse GPI-anchor fragments of T. brucei using a highly flexible sili-
con based protecting group strategy.9,12 The 1,1,3,3-tetraisopropyl-
1,3-disiloxan-1,3-diyl (TIPS) group was therein applied as a tempo-
rary protecting group which lead to a strategy for the preparation
of GPI-anchor fragments requiring less protecting group manipula-
tions but offering more synthetic flexibility. Furthermore, we could
show that the octyl-O- and S-glycosides of such fragments were
suitable substrates for studying in vitro the enzymatic glycosyla-
tion patterns of GPI-anchor fragments with trypanosomal
membrane fractions, and that octyl-S-glycosides were better
substrates for such enzymes.9 So far we have prepared octyl-O-
and S-glycosides related to GPI-anchor fragments CD and CDE9
(Fig. 1) and to fragments CF, C(F)D and C(F)DE,12 respectively. Here
we present our results for the similar chemical synthesis of a series
of tri- through pentasaccharide octyl-S-glycosides related to the
GPI-anchor fragments BCD, BC(F)D and BC(F)DE of T. brucei
(Fig. 1) which we intend to use as substrates for further studying
Despite the highly variant structure of the VSG of Trypanosoma,
Ferguson et al.4 could show that the variable proteins are anchored
in the cell membrane of the parasites by a unique invariable gly-
can-phosphatidyl-inositol (GPI-anchor). GPI anchors are con-
structed out of a conserved 6-phosphatidyl-Man-a1,2-Man-a1,6-
Man-a1,4-GlcNH2-a1,6-myo-inositol-1-phosphate core structure
⇑
Corresponding author. Tel.: +49 (0) 7071 29 78763; fax: +49 (0) 7071 29 5244.
E-mail address: Thomas.ziegler@uni-tuebingen.de (T. Ziegler).
0008-6215/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2011.08.001

R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2349
O
OH
O
NH-(VSG-Protein)
P
O
OH
O
HO
HO
E
HO
HO
HO
O
O
OOC(CH2)12CH3
OOC(CH2)12CH3
OH
D
O
O
HO
OH
HO
O
O
O
α-
D
-Gal-(1,2)-O
P
O
C
O
O
HO
OH
HO
O
H2N
HO
B
O
F
HO
α-D-Gal-(1,2)-α-D-Gal-(1,6)-O
OH
HO
A
Figure 1. VSG-GPI anchor of Trypanosoma brucei.
the glycosylation patterns of trypanosomal GPI-anchor fragments.
Octyl-S-glycosides were chosen not only because they had been
previously shown to be better substrates for that purpose but also
because they could be easily converted into glycosyl donors and
thus, enabling straightforward strategies for their synthesis.
and sodium hydride to afford 7 in 89%. Finally, the benzylidene
group in glycoside 7 was regioselectively reduced with sodium
cyanoborohydride using Garegg’s16 and Nilsson’s procedure17 to
afford 8 in 89% (Scheme 1).
Previously, we have efficiently used 1,1,3,3-tetraisopropyl-1,3-
disiloxane-1,3-diyl- (TIPS) protected glycosyl donors and acceptors
for the synthesis of various oligosaccharides applying the flexible
glycodesilylation strategy.9,12,18–22 Therefore, we also chose the
TIPS-protected imidate 12 for glycosylation of monosaccharide 8
here. Glycosyl donor 12, bearing a 4,6-O-TIPS group for allowing
the extension of the sugar chain at position 6, was prepared from
the known TIPS-protected octyl 1-thio-glucoside 9.12 The latter
was first acetylated to give 10 (91%), followed by selective hydro-
lysis of the thioether moiety with N-bromosuccinimide (NBS) in
aqueous acetone23 to give the mannose derivative 11 (75%), fol-
lowed by treatment with trichloroacetonitrile and sodium carbon-
ate24 to afford imidate 12 in 93% yield (Scheme 2).
Coupling of imidate 12 with glucosyl acceptor 8 proceeded
smoothly with TMSOTf in diethylether to afford disaccharide 13
in 86% yield. Next, the TIPS-group in 13 was regioselectively
opened with pyridine–HF complex18–20 giving disaccharide accep-
tor 14 in a virtually quantitative yield. Mannosylation of the latter
with benzobromomannose 15 under Koenigs–Knorr conditions
gave 16 in 89% yield which was subsequently desilylated with
Bu4NF to afford trisaccharide 17 in 76% yield. The deprotection of
saccharide 17 was performed in two steps. First, compound 17
was Zemplén deprotected (NaOMe in MeOH) to give 18 (93%). Dif-
ficulties were encountered, however, when 18 was hydrogenated
with various types of Pd-catalysts under various conditions (Pd
on charcoal, Pearlman’s catalyst, Pd-black, EtOH/AcOH/H2O,
2. Results and discussion
For the synthesis of the trisaccharide octyl-S-glycoside frag-
ment Man-
a stepwise approach starting from octyl 2-azido-3,6-di-O-benzyl-
2-deoxy-1-thio- -glucopyranoside (8). The latter was prepared
in five steps as follows. First, 1,3,4,6-tetra-O-acetyl-2-azido-2-
deoxy-b- -glucopyranose (1), easily accessible from pentaacetyl
a1,6-Man-a1,4-GlcNH2-aS-octyl (19) (BCD) we chose
a
-D
D
mannose in two step using Kovac’s carefully optimized
procedure,13,14 was reacted with octylthiotrimethylsilane (2) and
trimethylsilyl triflate (TMSOTf) according to Glaudemans’ proce-
dure15 to afford the octyl
a-
D-glucopyranoside 3 in 71% yield. A
small amount (7%) of the corresponding b- -glucopyranoside 4
D
was also obtained but could be easily separated by a single chro-
matography. The anomeric configurations of both 3 and 4 were
evident from their NMR spectra which showed vicinal coupling
constants for H-1 and signals for C-1 at J1,2 = 5.5 Hz and 83.3 ppm
for 3, and J1,2 = 10.2 Hz and 84.6 ppm for 4, respectively. Likewise,
the a-anomer 3 had a specific rotation of +147.2 while that of the
b-anomer 4 was ꢀ27.9. Next, compound 3 was Zemplén deacety-
lated to give intermediate 5 (92%) which afforded the 4,6-O-ben-
zylidene protected glucoside
6 (85%) upon treatment with
benzaldehyde dimethyl acetal and camphorsulfonic acid. Com-
pound 5 was then benzylated at position 3 with benzyl bromide
OAc
OAc
O
OH
O
C8H17SSiMe3 (2)
NaOMe
MeOH
O
TMSOTf
AcO
AcO
AcO
AcO
HO
HO
OAc
N3
N3
N3
SC8H17
SC8H17
1
3 α-anomer (71%)
5
(92%)
4
β-anomer (7%)
1) PhCH(OMe)2
2) BnBr, NaH
OBn
Ph
O
NaCNBH3
O
O
O
HO
BnO
RO
N3
N3
SC8H17
SC8H17
6
7
R=H (85%)
8
(89%)
R=Bn (89%)
Scheme 1. Preparation of glucosyl acceptor 8.

2350
R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
Si
Si
Si
O
Si
O
1) Ac2O
2) NBS
OBz
O
O
Si
Si
O
O
8
O
OBz
OBz
O
3) Cl3CCN
TMSOTf
OBn
O
O
AcO
O
O
O
AcO
HO
O
BnO
R
SC8H17
N3
10 R=SC8H17 (91%)
13
(86%)
9
SC8H17
11 R=OH (75%)
12
R=OC(=NH)CCl3 (93%)
OBz
OBz
Py(HF)x
O
BzO
BzO
BzO
BzO
OBz
O
Si
F
OH
O
OBz
BzO
O
OBz
O
15
Br
Si
O
O
RO
OBn
O
OBn
AcO
AcO
1) AgOTf
2) Bu4NF
O
O
O
BnO
BnO
N3
N3
SC8H17
SC8H17
14
(99%)
16 R=F-Si(i-Prop)2-O-Si(i-Prop)2- (89%)
17
R=H (76%)
NaOMe
MeOH
OH
OH
O
OH
OH
O
HO
HO
HO
HO
O
O
Na/NH3
OH
OH
O
O
HO
HO
HO
HO
OBn
O
OH
O
O
O
BnO
HO
N3
H2N
SC8H17
SC8H17
19
(50%)
18 (93%)
Scheme 2. Preparation of trisaccharide fragment BCD (19).
1000–100,000 hPa). In all cases, hydrogenation was either incom-
plete or not reproducible, or the formation of side products
(namely de-aminated and acetylated side products) was observed.
Reduction of the azido group and reductive debenzylation of 18
could finally be achieved under carefully optimized Birch reduction
conditions with sodium in THF and liquid ammonia25 (30 mol-
equiv Na, workup with NH4Cl) giving trisaccharide octyl-S-glyco-
side 19 in 50% yield. The moderate yield for the deprotection step
was due to the fact that three chromatographies on silica gel, Bio-
gel P2 and RP-C18 silica gel were required in order to obtain pure
19 (Scheme 2).
block through intramolecular glycosylation via prearranged glyco-
sides.26,27 However, previous experiments on the intramolecular
glycosylation of the prearranged disaccharide as shown in Figure
2 resulted in a rearrangement of the TIPS group from positions 4
and 6 to positions 3 and 4 in the manno-moiety, followed by galac-
tosylation of position 6.28
Since additional problems were anticipated for the TIPS-pro-
tected octyl 1-thio-mannoside building blocks which we intended
to use here, we chose a classical direct
of building block 9 (Scheme 3). It has been shown previously that
1,3-galactosylation of 4,6-TIPS-protected glucosides proceeds
a-selective galactosylation
a
For the preparation of the galactosylated fragment BC(F)D we
rather sluggishly giving only medium yield of the corresponding
first contemplated to construct a suitable Gal-
a1,3-Man building
disaccharides.9,29 Therefore, we have tested the condensation of 9
OBn
O
BnO
BnO
OBn
O
BnO
BnO
O
Si
O
O
O
SEt
DMTST or NIS
O
O
O
Si
O
O
O
O
Si
O
O
O
O
23-35%
O
O
HO
Si
OMe
OMe
Figure 2. Intramolecular galactosylation of a 4,6-TIPS-protected methyl mannoside according to Ref. 28.

R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2351
Si
O
Si
O
Si
Si
O
O
1) NBS
OBz
O
OBz
Activator
2) Cl3CCN
O
O
O
9
O
O
OBn
O
BnO
BnO
BnO
BnO
SC8H17
R
BnO
BnO
BnO
O
O
X
BnO
BnO
22
OBn
21
(61%)
OBn
20a X=β-SEt
R=OH (64%)
20b
X=β-S(=O)Et
23 R=OC(=NH)CCl3 (72%)
20c X=α-C(=NH)CCl3
20d
X=α-Cl
8
TMSOTf
Si
OH
OBz
F
Si
O
Si
Si
O
O
OBz
O
O
O
O
OBn
Py(HF)x
O
BnO
O
OBn
O
O
O
BnO
BnO
BnO
O
O
BnO
BnO
N3
SC8H17
BnO
O
BnO
OBn
N3
SC8H17
OBn
24
25
(89%)
(91%)
15, AgOTf
then Bu4NF
OH
OBz
OBz
OH
O
O
HO
HO
BzO
BzO
O
O
NaOMe then
Na, NH3
OH
OBz
O
O
HO
HO
O
OH
O
O
OBn
O
HO
BnO
O
O
HO
HO
BnO
BnO
HO
BnO
O
O
H2N
N3
OH
OBn
SC8H17
SC8H17
27
26 (68%)
(65%)
Scheme 3. Preparation of tetrasaccharide fragment BC(F)D (27).
with various fully benzyl-protected galactosyl donors (ethyl 1-
thio-b- -trichloroacetimidate
-galactoside 20a,30 sulfoxide 20b,
20c31 and
tion or gave incomplete reaction and only medium yields of the
desired disaccharide 21. Sulfoxide 20b and imidate 20c (Table 1,
entries 6 and 7) did not afford disaccharide 21 at all. Best results
D
a
a
-chloride 20d32) and various activation procedures
(AgOTf/Br2,33 MeOTf,34 NIS/TMSOTf,35 iodoniumdicollidine per-
chlorate (IDPC),36 Tf2O,37 TMSOTf, AgClO429). Table 1 summarizes
the results for these glycosylation reactions. Coupling with S-gly-
coside 20a (Table 1, entries 1–5) either resulted in its decomposi-
were obtained for the a1,3-selective galactosylation of 9 with chlo-
ride 20d under carefully optimized reaction conditions (Table 1,
entries 8 and 9) affording 21 in 61% yield (Scheme 3). The anomeric
configuration was evident from the NMR spectrum of 21 which
Table 1
Condensation of 9 and 20a–d under various conditions (see Scheme 3)
Entry
20a–d
9:20a–da
Solvent
Activator
Condition
21
9b
1
2
3
4
5
6
7
8
20a
20a
20a
20a
20a
20b
20c
20d
1:1.3
1:1.3
1:2.0
1:1.3
1:1.3
1:1
Toluol
CH2Cl2
Et2O
CH2Cl2
AgOTf/Br2
AgOTf/Br2
MeOTf
0 °C?rt, 2 h
0 °C?rt, 2 h
rt, 24 h
rt
46%
Decomp.
18%
Decomp.
Decomp.
Decomp.
Decomp.
51%
13%
42%
NIS/TMSOTf
CH2Cl2
IDCPc
rt
CH2Cl2/Tol
CH2Cl2/Et2O
CH2Cl2
Tf2O/s-Coll.
TMSOTf
AgClO4
s-Coll. 2 equiv
AgClO4
s-Coll. 11 equiv
ꢀ50 °C
1:1
1:1.2
ꢀ30 °C
0 °C?rt, 2 h
—
—
9
20d
1:1.2
CH2Cl2
ꢀ5 °C?rt, 1.5 h
61%
a
b
c
Mol-ratio.
Reisolated starting material.
Iodoniumdicollidine perchlorate.

2352
R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
showed a vicinal coupling constant J1,2 for H-1 of the galactosyl
residue of 3.5 Hz.
sis of BF3 (glycodesilylation).19–22 Therefore, we also tested
whether disaccharide 21 can be regioselectively mannosylated
with benzofluoromannose 32 at position 6. Fluoride 32 was pre-
pared in 81% from benzobromomannose 15 and KHF2 pursuant
to Thiem’s procedure.38 However, when 21 was reacted with 32
and BF3-etherate in dichloromethane, only the galactosylated
1,6-anhydro-mannose 33 and octyl 2,3,4,6-tetra-O-benzoyl-1-
Disaccharide 21 was converted into the corresponding trichlo-
roacetimidate donor 23 essentially as described above for mono-
saccharide donor 12. S-Glycoside 21 was hydrolyzed with NBS to
give 22 (64%) followed by treatment with trichloroacetonitrile to
afford 23 in 72% yield (Scheme 3). Next, the latter was condensed
with acceptor 8 to give the corresponding trisaccharide 24 in excel-
lent 91% yield. Regioselective fluorodesilylation of the TIPS-group
with pyridine–HF gave 60-OH trisaccharide 25 (89%) which was
mannosylated with benzobromomannose 15 under Koenigs–Knorr
conditions and desilylated to afford tetrasaccharide 26 (68%) in
two steps. Finally, sequential deprotection of the latter (Zemplén
deacylation followed by reduction of the benzyl and azido groups
with Na in liquid ammonia) gave the free tetrasaccharide 27 in
65% yield.
In an alternative approach to tetrasaccharide 26 we prepared a
trisaccharide donor as follows. Instead of using disaccharide 21 as
the donor (see Scheme 3), we now converted it into a suitable
acceptor by once again regioselectively hydrolyzing the 4,6-O-TIPS
group in 21 with the pyridine–HF complex (Scheme 4). Thus ob-
tained disaccharide acceptor 28 (95%) was mannosylated with
benzobromomannose to give trisaccharide 29 in 73% yield. Since
some characteristic signals in the NMR spectra of 29 were over-
lapped and thus, could not get unambiguously assigned, 29 was
desilylated to 30 (89%) the NMR spectra of which now could be
fully interpreted. Next, 29 was converted in 95% yield into the cor-
responding trisaccharide imidate donor 31 by sequential hydroly-
sis of the aglycon with NBS followed by treatment with
trichloroacetonitrile. Glycosylation of 8 with the latter proceeded
somewhat sluggishly though. Tetrasaccharide 26 was only ob-
tained in medium 44% yield. Nevertheless, both strategies as out-
thio-a-D-mannopyranoside (34) could be isolated in 50% and 64%
yield, respectively. Obviously, the intermediate mannosyl cation
attacked the anomeric sulfur in 21 to give intermediates A and B
(Scheme 5) which subsequently led to the products 33 and 34. Sim-
ilar intermolecular aglycon transfers had been observed upon gly-
cosylations of thioglycosides with glycosyl imidates and
halogenoses.39–42
Trisaccharide 25 was also used for the preparation of the octyl
S-glycoside pentasaccharide fragment related to GPI-anchor
BC(F)DE as follows. Condensation of 25 with disaccharide imidate
3512 followed by fluoride-catalyzed desilylation of the intermedi-
ate afforded the pentasaccharide 36 in 86% overall yield (Scheme
6). Sequential Zemplén deprotection and Birch reduction of the lat-
ter then gave 37 in 53% yield over two steps.
In summary, we have efficiently prepared three octyl-S-glyco-
side tri-, tetra- and pentasaccharide fragments (19, 27, 37) related
to the GPI-anchor of trypanosoma brucei using TIPS-protected gly-
cosylacceptors. The three octyl-S-glycosides will be used as sub-
strates for trypanosomal glycosidases and glycosyl transferases in
order to further study the glycosylation pattern of trypanosoma
brucei as previously described for similar GPI-anchor related oc-
tyl-S-glycosides.9
3. Experimental
lined in Schemes
preparation of the GPI-anchor fragment BC(F)D.
3 and 4 are likewise suitable for the
3.1. General methods
1H and 13C NMR spectra were recorded with Bruker ARX 250,
Avance 400 or AMX 600 spectrometers at 250, 400 or 600 MHz
Previously we had shown that TIPS-protected glycosides can get
regioselectively glycosylated with glycosyl fluorides under cataly-
BzO
OBz
O
OH
F
BzO
OBz
BzO
Si
Si
O
O
O
O
O
OBz
Py(HF)x
15, AgOTf
F
Si
BnO
21
SC8H17
Si
O
BnO
O
O
O
O
BnO
OBn
BnO
SC8H17
BnO
BnO
29
28 (95%)
O
OBn
(73%)
NBS then
Cl3CCN
Bu4NF
BzO
BzO
BzO
OBz
O
OBz
O
BzO
BzO
BzO
8
, TMSOTf
O
OBz
O
OBz
F
Si
then Bu4NF
Si
26
(44%)
O
O
O
O
HO
NH
O
O
BnO
BnO
O
CCl3
SC8H17
BnO
BnO
BnO
BnO
O
O
OBn
OBn
31 (95%)
30 (89%)
Scheme 4. Alternative preparation of tetrasaccharide fragment BC(F)D.

R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2353
OBz
Si
OBz
OBn
O
BnO
BnO
OBz
O
O
Si
BzO
BzO
O
F
OBz
OBz
O
O
32
O
BzO
BzO
BnO
+
O
O
BnO
O
SC8H17 BF3 then Bu4NF
SC8H17
BnO
BnO
34 (64%)
BzO
O
O
OH
21
OBn
33 (50%)
F
BF3
Si
O
Si
F
O
O
Si
O
Si
O
OBz
OBz
O
O
O
O
O
BnO
BnO
SC8H17
SC8H17
BnO
BnO
BnO
BnO
OBz
OBz
OBz
O
OBz
OBz
OBz
O
O
O
OBz
OBn
OBn
B
A
OBz
Scheme 5. Glycosylation of disaacharide acceptor 21 with mannosyl donor 32.
OBz
OBz
OH
OBz
O
OBz
OH
O
BzO
BzO
BzO
O
HO
HO
BzO
AcO
AcO
AcO
O
AcO
AcO
AcO
O
HO
HO
HO
O
O
O
O
NH
CCl3
NaOMe, MeOH
then Na, NH3
35
O
O
O
OBz
25
OH
O
HO
O
TMSOTf then Nu4NF
HO
OBn
O
O
OH
O
BnO
HO
O
O
BnO
BnO
O
BnO
HO
HO
O
HO
O
N3
SC8H17
H2N
OBn
OH
SC8H17
36 (86%)
37 (53%)
Scheme 6. Preparation of pentasaccharide fragment BC(F)DE (37).
for protons and 62.9, 100 or 151 MHz for carbons, respectively.
Chemical shifts in CDCl3 are reported in d (ppm) relative to tetra-
methylsilane (TMS) as internal standard. Coupling constants J are
reported in Hertz. Assignment of signals was made by first order
inspection of the spectra and, if necessary, by 1H1H-, 13C1H-COSY,
HMQC or NOESY experiments, respectively. MS spectra were re-
corded with Finnigan MAT TSQ 70 (EI, CI), Finnigan MAT 711 A
(HRMS), Bruker Autoflex (MALDI-TOF-MS) and Bruker Apex II FT-
filtered and concentrated with a rotary evaporator. HPLC was per-
formed on a Sykam system using a Grom Saphir Si; 5 lm;
250 ꢁ 6 mm column.
3.2. Octylthiotrimethylsilane (2)
A solution of thiooctanol (70 mL, 403 mmol) in n-hexane
(100 mL) was dropped at rt with vigorous stirring into a slurry of
Na pieces (9.34 g, 406 mmol) in n-hexane (100 mL), the mixture
was refluxed for 2 h and cooled to rt. A solution of trimethylchlo-
rosilane (51 mL, 403 mmol) in n-hexane (80 mL) was added drop-
wise with stirring, the mixture refluxed for another 2 h and set
aside at rt without stirring for 24 h. Filtration of the mixture, con-
centration of the filtrate and distillation of the residue gave 2 as a
colorless liquid (58.46 g, 67%). Bp 147 °C (50 hPa). 1H NMR (CDCl3):
d 2.48 (t, 2H, J = 7.2 Hz, SCH2), 1.64–2.55 (m, 2H, CH2), 1.40–1.27
(m, 10H, CH2), 0.88 (t, 3H, J = 6.2 Hz, CH3), 0.31 (br t, 9H,
J = 3.3 Hz, Si(CH3)3). 13C NMR (CDCl3): d 33.1, 31.8, 29.2, 29.1,
28.8, 26.4, 22.6 (CH2), 14.1 (CH3), 0.9 (3C, Si(CH3)3). Anal. Calcd
for C11H26SSi (218.48): C, 60.48; H, 11.99; S, 14.67. Found: C,
60.32; H, 11.94; S, 14.86.
ICR (FAB-MS, HRMS) instruments. Specific optical rotations [a]
were recoded with a Perkin Elmer polarimeter Model 341 at
589 nm (Na-D) for solutions in CHCl3 at 20 °C if not stated other-
wise. Elemental analyses were performed with a Hekatech CHNS
analysator Euro EA 3000. Melting points were determined on a
Büchi B-540 instrument. TLC was performed on Macherey & Nagel
silica gel SIL G/UV254 plates. Spots were detected by visual inspec-
tion of the plates under UV light, by charring with H2SO4 (5% in
EtOH), with KMnO4 (5% in water) and with I2, respectively. Pre-
parative column chromatography was performed by eluting com-
pounds with various mixtures of solvents from glass columns of
various sizes filled with Macherey & Nagel silica gel S (0.032–
0.063 mm). Solutions in organic solvents were dried with Na2SO4,

2354
R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
3.3. Octyl 2-azido-3,4,6-tri-O-acetyl-2-deoxy-1-thio-
glucopyranoside (3) and octyl 2-azido-3,4,6-tri-O-acetyl 2-
deoxy-1-thio– -glucopyranoside (4)
a
-
D
-
d
7.52–7.10 (m, 5H, PhH), 5.52 (s, 1H, PhCH), 5.32 (d, 1H,
J1,2 = 5.6 Hz, H-1), 4.28–4.19 (m, 2H), 3.96 (t, 1H, J = 9.6 Hz), 3.80
(dd, 1H, J2,3 = 10.0 Hz, H-2), 3.74 (t, 1H, J = 11.6 Hz), 3.49 (t, 1H,
J = 9.0 Hz), 3.03 (s, 1H, OH), 2.65–2.49 (m, 2H, SCH2), 1.67–1.57
(m, 2H, CH2), 1.49–1.27 (m, 10H, CH2), 0.90–0.86 (br t, 3H, CH3).
13C NMR (CDCl3): d 136.7, 129.4, 128.6, 125.7 (PhC), 102.1 (PhCH),
84.3 (C-1), 81.8 (C-4), 70.5 (C-3), 68.6 (C-6), 63.7 (C-5), 62.8 (C-2),
31.7, 30.9, 29.5, 29.1 (2C), 28.8, 22.6 (CH2), 14.0 (CH3). Anal. Calcd
for C21H31N3O7S (421.56): C, 59.83; H, 7.41; N, 9.97; S, 7.61. Found:
C, 59.57, H, 7.44; N, 9.90; S, 7.76.
D
A solution of 2-azido-2-deoxy-1,3,4,6-tetra-O-acetyl-b-
pyranoside (1)13 (1.0 g, 2.67 mmol), compound
6.95 mmol) and a catalytic amount of TMSOTf (100
D
-gluco-
(1.9 mL,
L, 0.5 mmol)
2
l
in dichloroethane (40 mL) was stirred at 70 °C for 14 d. After the
mixture was cooled to rt, it was diluted with dichloromethane
(40 mL), washed with saturated aq NaHCO3 solution, dried, filtered
and concentrated. Chromatography of the residue with 5:1 n-hex-
ane/ethyl acetate gave first 3 (0.87 g, 71%) as colorless oil. [
a]
3.6. Octyl 2-azido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-1-
D
+147.2 (c 1.5, CHCl3), IR k (cmꢀ1) 2108.73 (N3). 1H NMR (CDCl3):
d 5.42 (d, 1H, J1,2 = 5.5 Hz, H-1), 5.29 (dd, 1H, J2,3 = 10.5 Hz,
J3,4 = 9.2 Hz, H-3), 5.01 (dd, 1H, J4,5 = 10.1 Hz, H-4), 4.45 (ddd, 1H,
J5,6a = 4.4 Hz, J5,6b = 2.0 Hz, H-5), 4.29 (dd, 1H, J6a,6b = ꢀ12.3 Hz, H-
6a), 4.06 (dd, 1H, H-6b), 3.98 (dd, 1H, H-2), 2.67–2.51 (m, 2H,
SCH2), 2.08 (s, 3H, CH3), 2.07 (s, 3H, CH3), 2.04 (s, 3H, CH3).1.73–
1.59 (m, 2H, CH2), 1.41–1.27 (m, 10H, CH2), 0.88 (br t, 1H,
J = 6.7 Hz, CH3). 13C NMR (CDCl3): d 170.5, 169.8 (2C, C@O), 83.3
(C-1), 72.1 (C-3), 68.7 (C-4), 67.9 (C-5), 91.9 (C-6), 61.6 (C-2),
31.8, 30.7, 29.4, 29.1 (2C), 28.9, 22.6 (CH2), 20.6, 20.7 (2C), 14.1
(CH3). Anal. Calcd for C20H33N3O4S (459.56): C, 52.27; H, 7.24; N,
9.14; S, 6.98. Found: C, 52.24; H, 7.29; N, 9.13; S, 7.44.
thio-a-D-glucopyranoside (7)
NaH (1.15 g, 48 mmol) was added in small portions at 0 °C to a
solution of compound 6 (10.14 g, 24.1 mmol) and benzyl bromide
(4.5 mL, 38 mmol) in DMF (100 mL) and stirred at rt for 1 h. Water
was carefully added dropwise to destroy the excess of NaH and the
resulting solution was poured into an ice/water mixture. The solid
material which separated was filtered off, washed with water and
dissolved in dichloromethane (100 mL). The organic solution was
washed with cold 1 M aq HCl solution, dried, filtered and concen-
trated. Crystallization of the residue from n-hexane gave
(10.95 g, 89%) as white crystals. Mp 69 °C. [ D +63.5 (c 1.3, CHCl3).
7
a]
Eluted next was 4 (90 mg, 7%) as a colorless oil. [
a]
D
ꢀ27.9 (c
1H NMR (CDCl3): d 7.51–7.24 (m, 10H, PhH), 5.58 (s, 1H, PhCH),
5.33 (d, 1H, J1,2 = 5.0 Hz, H-1), 4.93 (d, 1H, J = ꢀ10.9 Hz, PhCH2),
4.78 (d, 1H, J = ꢀ10.9 Hz, PhCH2), 4.95–4.23 (m, 2H), 3.95–3.67
(m, 4H), 2.66–2.50 (m, 2H, SCH2), 1.67–1.57 (m, 2H, CH2), 1.41–
1.27 (m, 10H, CH2), 0.88 (br t, 1H, J = 6.7 Hz, CH3). 13C NMR (CDCl3):
d 137.7, 137.1, 129.0, 128.4, 128.2, 128.1, 127.9, 126.0 (PhC), 101.4
(PhCH), 84.5 (C-1), 82.8 (C-4), 77.8 (C-3), 75.1 (PhCH2), 68.7 (C-6),
63.4 (C-5), 63.2 (C-2). Anal. Calcd for C28H37N3O4S (511.683): C,
65.73; H, 7.29; N, 8.21; S, 6.27. Found: C, 66.04; H, 7.31; N, 8.21;
S, 6.32.
1.2, CHCl3). 1H NMR (CDCl3): d 5.07 (t, 1H, J2,3 = J3,4 = 9.4 Hz, H-3),
5.01 (t, 1H, J3,4 = J4,5 = 9.5 Hz, H-4), 4.37 (d, 1H, J1,2 = 10.2 Hz, H-
1), 4.25 (dd, 1H, J5,6a = 5.2 Hz, J6a,6b = ꢀ12.3 Hz, H-6a), 4.11 (dd,
1H, J5,6b = 2.4 Hz, H-6b), 3.67 (ddd, 1H, H-5), 3.52 (t, 1H, H-2),
2.78–2.68 (m, 2H, SCH2), 2.10 (s, 3H, CH3), 2.07 (s, 3H, CH3), 2.02
(s, 3H, CH3), 1.71–1.60 (m, 2H, CH2), 1.42–1.27 (m, 10H, CH2),
0.90–0.86 (m, 3H, CH3). 13C NMR (CDCl3): d = 170.5, 169.9, 169.5
(C@O), 84.6 (C-1), 75.7 (C-5), 74.4 (C-3), 68.2 (C-4), 63.6 (C-2),
62.1 (C-6), 31.7, 30.8, 29.7, 29.1, 29.0, 28.7, 22.6 (CH2), 20.6 (2C),
20.5, 14.0 (CH3). Anal. Calcd for C20H33N3O7S (459.56): C, 52.27;
H, 7.24; N, 9.14; S, 6.98. Found: C, 52.35; H, 7.23; N, 9.04; S, 7.24.
3.7. Octyl 2-azido-3,6-di-O-benzyl-2-deoxy-1-thio-a-D-
glucopyranoside (8)
3.4. Octyl 2-azido-2-deoxy-1-thio-a-D-glucopyranoside (5)
A dry solution of HCl in diethylether was added dropwise with
stirring at rt to a solution of 7 (5.73 g, 11.2 mmol) and NaCNBH3
(4.23 g, 67.4 mmol) in THF (150 mL) until the solution became
acidic toward litmus. Stirring was continued for 10 min, the result-
ing suspension diluted with dichloromethane (ca. 100 mL) and fil-
tered through a layer of Celite. The filtrate was successively
washed with satd aq NaHCO3 solution, 1 M aq FeCl2 solution and
water, dried and concentrated. Chromatography of the residue
with 25:1 toluene/ethyl acetate gave 8 (5.1 g, 89%) as a colorless
A 1 M solution of NaOMe in MeOH (ca. 0.1 mL) was added at rt
to a solution of 4 (18.5 g, 41.7 mmol) in MeOH (100 mL) and the
resulting solution was stirred for 24 h. Neutralization of the solu-
tion with ion exchange resin (Dowex 50XW8, H+ form), filtration
and concentration of the filtrate gave a white crystalline solid
which was recrystallized from diethylether/n-hexane to afford 5
(11.33 g, 82%). Mp 89–90 °C. [a]
+208.9 (c 1.1, MeOH). 1H NMR
D
(MeOD-d3): d 5.36 (d, 1H, J1,2 = 4.6 Hz, H-1), 3.96–3.91 (m, 1H),
3.81–3.58 (m, 4H), 3.41–3.31 (m, 1H), 2.67–2.51 (m, 2H, SCH2),
1.67–1.53 (m, 2H, CH2), 1.40–1.29 (m, 10H, CH2), 0.90–0.87 (m,
3H, CH3). 13C NMR (MeOD-d3): d 84.9 (C-1), 74.4 (C-3), 74.1 (C-
4), 71.9 (C-5), 71.7 (C-2), 62.3 (C-6), 32.9, 31.3, 30.5, 30.3, 30.2,
29.9, 23.7 (CH2), 14.4 (CH3). Anal. Calcd for C14H27N3O4S
(333.45): C, 50.43; H, 8.16; N, 12.60; S, 9.62. Found: C, 50.52; H,
8.08; N, 12.48; S, 9.62.
oil. [a]
+91.3 (c 1.1, CHCl3). 1H NMR (CDCl3): d 7.41–7.24 (m,
D
10H, PhH), 5.33 (d, 1H, J1,2 = 5.5 Hz, H-1), 4.89 (d, 1H,
J = ꢀ11.1 Hz, PhCH2), 4.78 (d, 1H, J = ꢀ11.1 Hz, PhCH2), 4.59 (d,
1H, J = 12.1 Hz, PhCH2), 4.51 (d, 1H, J = 12.0 Hz, PhCH2), 4.16–4.12
(m, 1H, H-5), 3.81 (dd, 1H, J2,3 = 9.8 Hz, H-2), 3.76–3.59 (m, 4H,
H-3, H-4, H-6a,b), 2.63–2.51 (m, 2H, SCH2), 2.35–2.14 (m, 2H,
CH2), 1.63–1.62 (m, 10H, CH2), 0.90–0.86 (m, 3H, CH3). 13C NMR
(CHCl3): d 137.9, 137.7, 128.6, 128.4, 128.1, 128.0, 127.7, 127.6
(PhC), 83.6 (C-1), 81.3 (C-3), 75.3 (PhCH2), 73.6 (PhCH2), 72.1 (C-
4), 70.4 (C-5), 69.6 (C-6), 62.4 (C-2), 31.7, 30.6, 29.5, 29.1, 28.8,
22.6 (CH2), 14.0 (CH3). Anal. Calcd for C28H39N3O4S (513.70): C,
65.47; H, 7.65; N, 8.18; S, 6.24. Found: C, 65.46; H, 7.57; N, 7.98;
S, 6.06.
3.5. Octyl 2-azido-4,6-O-benzylidene-2-deoxy-1-thio-a-D-
glucopyranoside (6)
A solution of compound 5 (2.5 g, 7.5 mmol), benzaldehyde
dimethylacetal (4.2 mL, 28.0 mmol) and a catalytic amount of cam-
phorsulfonic acid (ca. 10 mg) in MeCN (40 mL) was stirred at rt for
3 h, neutralized by the addition of Et3N (1 mL) and concentrated.
Trituation of the oily reside with n-hexane (50 mL) gave white
crystals which were filtered off, washed with a small amount of
cold n-hexane and dried. Recrystallization with n-hexane gave 6
3.8. Octyl 3-O-acetyl-2-O-benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-
1,3-disiloxane-1,3-diyl)-1-thio-a-D-mannopyranoside (10)
Ac2O (3 mL) was added at 0 °C to a solution of compound 912
(0.82 g, 1.25 mmol) and a catalytic amount of DMAP (10 mg) in
(2.68 g, 85%). Mp 95 °C. [
a
]
D +164.4 (c 1.1, CHCl3). 1H NMR (CDCl3):

R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2355
pyridine (6 mL), and the mixture was stirred at rt for 16 h. The mix-
3.11. Octyl 3-O-acetyl-2-O-benzoyl-4,6-O-(1,1,3,3-
tetraisopropyl-1,3-disiloxane-1,3-diyl)- -mannopyranosyl-
(1?4)-2-azido-3,6-di-O-benzyl-2-deoxy-1-thio-a-D-
ture was poured into ice-cold diluted aqueous HCl solution (ca.
300 mL) and extracted with dichloromethane (3 ꢁ 50 mL). The
combined extracts were successively washed with diluted aqueous
HCl solution, saturated NaHCO3 solution, dried, filtered and con-
centrated. Chromatography of the residue with toluene gave 10
a-D
glucopyranoside (13)
TMSOTf (55
lL, 0.3 mmol) was added under Ar at ꢀ15 °C to a
(0.79 g, 91%) as a colorless oil. [
a]
+16.9 (c 0.9, CHCl3). 1H NMR
suspension solution of compound 12 (1.28 g, 1.79 mmol), com-
pound 8 (0.78 g, 1.52 mmol) and 4 Å molecular sieves (5.0 g) in
diethylether (100 mL). The mixture was warmed to 0 °C within
1.5 h, NaHCO3 (ca. 1 g) was added, and the mixture was filtered
through a layer of celite. The filtrate was washed with aqueous
NaHCO3 solution, dried, filtered and concentrated. Chromatogra-
phy of the residue with 25/1 toluene–ethyl acetate gave 13
D
(CDCl3): d 8.08–7.15 (m, 5H, PhH), 5.63 (dd, 1H, J1,2 = 1.2 Hz,
J2,3 = 3.3 Hz, H-2), 5.40 (d, 1H, H-1), 5.28 (dd, 1H, J3,4 = 9.9 Hz, H-
3), 4.49 (t, 1H, J = 9.4 Hz, H-4), 4.24 (dd, 1H, J5,6a = 2.0 Hz,
J6a,6b = 12.6 Hz, H-6a), 4.11 (br d, 1H, J = 9.4 Hz, H-5), 3.92 (dd,
1H, J5,6b = 0.5 Hz, H-6b), 2.60 (dddd, 2H, J = 29.7 Hz, J = 12.9 Hz,
J = 7.3 Hz, J = 7.3 Hz, S-CH2), 1.98 (s, 3H, Acetyl-CH3), 1.64–0.83
(m, 42H, CH, CH2, CH3). 13C NMR (CDCl3): d 170.0, 165.5 (C@O),
133.3, 129.9, 129.0, 128.3, 128.2, 125.3 (PhC), 83.1 (C-1), 73.7 (C-
5), 72.4 (C-3), 72.3 (C-2), 64.8 (C-4), 61.0 (C-6), 31.8, 31.3, 29.5,
29.1 (2C), 28.8, 22.6 (CH2), 21.0, 17.4, 17.2 (5C), 14.0, 13.5, 13.2,
12.5, 12.4 (CH, CH3). Anal. Calcd for C35H60O8SSi2 (697.08): C,
60.30; H, 8.68. Found: C, 60.18; H, 8.46.
(1.39 g, 86%) as a colorless oil. [
a]
+28.2 (c 0.5, CHCl3). 1H NMR
D
0
0
(CDCl3): d 7.94–6.92 (m, 15H, PhH), 5.57 (dd, 1H, J1 ,2 = 1.5 Hz,
J2 ,3 = 3.2 Hz, H-2‘), 5.49 (br s, 1H, H-10), 5.41 (d, 1H, J1,2 = 4.9 Hz,
0
0
H-1), 5.26 (dd, 1H, J3 ,4 = 9.9 Hz, H-30), 4.90 (d, 1H, J = ꢀ10.5 Hz,
PhCH2), 4.70 (d, 1H, J = ꢀ10.9 Hz, PhCH2), 4.56 (br s, 2H, PhCH2),
0
0
4.33 (t, 1H, J3 ,4 = J4 ,5 = 9.8 Hz, H-40), 4.30–4.24 (m, 1H, H-5), 3.93
(t, 1H, J3,4 = J4,5 = 8.3 Hz, H-4), 3.88–3.57 (m, 7H, H-2, H-3, H-50,
H-6a,b, H-60a,b), 2.67–2.55 (m, 2H, SCH2), 1.97 (s, 3H, Acetyl-
CH3), 1.68–1.57 (m, 2H, CH2), 1.38–1.27 (m, 12H, CH2), 1.06–0.81
(m, 29H, CH, CH3). 13C NMR (CDCl3): d 170.3, 165.2 (C@O), 138.0,
137.1, 133.2, 129.9, 129.0, 128.3, 128.2, 128.1, 127.6, 127.4, 127.1
(PhC), 99.1 (C-10), 83.5 (C-1), 82.0 (C-3), 75.2 (PhCH2), 74.6 (C-4),
74.1 (C-50), 73.4 (PhCH2), 71.9 (C-30), 70.5 (C-20), 70.4 (C-5), 68.9
(C-6), 64.5 (C-2), 64.2 (C-40), 60.6 (C-60), 31.8, 30.6, 29.6, 29.1,
28.9, 22.6 (CH2), 21.0 (Acetyl-CH3), 17.4, 17.3, 17.2, 17.1, 17.0,
14.1, 13.4, 13.1, 12.5, 12.3 (CH, CH3). Anal. Calcd for C55H81N3O12S-
Si2 (1064.48): C, 62.02; H, 1.67; N, 3.95; S, 3.01. Found: C, 62.27; H,
7.75; N, 3.61; S, 3.07.
0
0
0
0
3.9. 3-O-Acetyl-2-O-benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-
disiloxane-1,3-diyl)-a-D-mannopyranose (11)
NBS (0.74 g, 4.13 mmol) was added at 0 °C with stirring to a
solution of compound. 10 (1.77 g, 2.54 mmol) in a mixture of ace-
tone (40 mL) and water (0.4 mL) and the mixture was stirred for
10 min. Saturated aqueous NaHCO3 solution (1 mL) was added,
the mixture was poured into diluted aqueous NaHCO3 solution
(100 mL) and extracted with dichloromethane (3 ꢁ 25 mL). The
combined organic extracts were washed with water, dried, filtered
and concentrated. Chromatography of the residue with 10/1 tolun-
ene–ethyl acetate gave 11 (1.09 g, 76%) as a colorless oil. [a]
D
ꢀ65.4 (c 1.3, CHCl3). 1H NMR (CDCl3): d 8.12–7.26 (m, 5H, PhH),
5.57 (dd, 1H, J1,2 = 1.6 Hz, J2,3 = 3.2 Hz, H-2), 5.41 (dd, 1H,
J3,4 = 10.0 Hz, H-3), 5.37 (dd, 1H, J1,OH = 3.3 Hz, H-1), 4.46 (t, 1H,
J = 9.8 Hz, H-4), 4.21 (dd, 1H, J5,6a = 1.9 Hz, J6a,6b = 12.6 Hz, H-6a),
3.99 (br d, 1H, J = 9.4 Hz, H-5), 3.92 (dd, 1H, J5,6b = 0.9 Hz, H-6b),
3.30 (d, 1H, OH), 2.00 (s, 3H, Acetyl-CH3), 1.15–0.97 (m, 28H, CH,
CH3). 13C NMR (CDCl3): d 170.3, 165.7 (C@O), 133.3, 129.4, 128.9,
128.3 (PhC), 92.9 (C-1), 73.2 (C-5), 71.8 (C-3), 70.9 (C-2), 64.5 (C-
4), 61.0 (C-6), 21.0 (CH3), 17.4, 17.3, 17.2 (2C), 17.1 (2C),13.5,
13.2, 12.5, 12.4 (CH, CH3). Anal. Calcd for C27H44O9Si2 (568.80): C,
57.01; H, 7.80. Found: C, 56.82; H, 7.79.
3.12. Octyl 3-O-acetyl-2-O-benzoyl-4-O-(1-fluoro-1,1,3,3-
tetraisopropyl-1,3-disiloxane-3-yl)-a-D-mannopyranosyl-
(1?4)-2-azido-3,6-di-O-benzyl-2-deoxy-1-thio-a-D-
glucopyranoside (14)
Pyridine hydrofluoride (70%, 127 lL, 4.45 mmol) was added at
rt to a stirred solution of compound 13 (0.68 g, 0.65 mmol) in
dichloromethane (20 mL) and stirring continued for 5 min. The
mixture was poured into diluted aqueous NaHCO3 solution (ca.
150 mL) and extracted with dichloromethane (3 ꢁ 25 mL). The
combined organic extracts were dried, filtered and concentrated.
Chromatography of the residue with 20/1 toluene–ethyl acetate
3.10. 3-O-Acetyl-2-O-benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-
gave 14 (0.69 g, 99%) as a colorless oil. [
a] +47.7 (c 0.9, CHCl3).
D
1H NMR (CDCl3): d7.95–7.02 (m, 15H, PhH), 5.62 (dd, J1 ,2 = 2.0 Hz,
0
0
disiloxane-1,3-diyl)-a-D-mannopyranosyl trichloroacetimidate
J2 ,3 = 3.0 Hz, H-20), 5.42 (d, 1H, H-10), 5.41 (d, 1H, J1,2 = 4.7 Hz, H-1),
0
0
(12)
5.19 (dd, 1H, J3 ,4 = 9.6 Hz, H-30), 4.91 (d, 1H, J = ꢀ10.6 Hz, PhCH2),
4.76 (d, 1H, J = ꢀ10.6 Hz, PhCH2), 4.61 (d, 1H, J = ꢀ12.2 Hz, PhCH2),
0
0
A mixture of compound 11 (1.09 g, 1.92 mmol), K2CO3 (2.06 g
14.9 mmol) and trichloroacetonitrile (2.1 mL, 20.9 mmol) in
dichloromethane (50 mL) was stirred at rt for 16 h, centrifugated,
and the centrifugate was concentrated. Chromatography of the res-
idue with 30/1 toluene–ethyl acetate gave 12 (1.28 g, 93%) as a yel-
0
0
0
0
4.57 (d, 1H, J = ꢀ12.2 Hz, PhCH2), 4.34 (t, 1H, J3 ,4 = J4 ,5 = 9.3 Hz, H-
40), 4.26 (br d, 1H, J = 8.5 Hz, H-5), 3.98 (t, 1H, J3,4 = J4,5 = 9.0 Hz, H-
4), 3.92 (dd, 1H, J6a,5 = 3.6 Hz, J6a,6b = ꢀ11.4 Hz, H-6a), 3.88–3.75
(m, 5H, H-2, H-3, H-50, H-6a,b0), 3.69 (dd, 1H, J6b,5 = 1.8 Hz, H-6b),
2.65–2.54 (m, 2H, SCH2), 2.04 (t, J = 6.2 Hz, OH), 1.98 (s, 3H, Acet-
yl-CH3), 1.69–1.60 (m, 2H, CH2), 1.38–1.02 (m, 11H, CH2), 1.01–
0.87 (m, 31H, CH,CH3). 13C NMR (CDCl3): d 170.3, 165.1 (C@O),
138.0, 137.1, 133.3, 129.7, 129.4, 129.0, 128.4. 128.3, 128.2 (2C),
127.9, 127.6 (2C), 127.4, 125.3 (PhC), 98.9 (C-10), 83.4 (C-1), 81.7
(C-3), 75.5 (C-4), 75.4 (PhCH2), 74.3 (C-50), 73.5 (PhCH2), 72.5 (C-
30), 70.7 (C-5), 70.0 (C-20), 68.5 (C-6), 65.6 (C-40), 64.6 (C-2), 61.5
(C-60), 31.8, 30.7, 29.5, 29.1 (2C), 28.9, 22.6 (CH2), 20.9 (Acetyl-
CH3), 17.2, 17.1 (3C), 16.6, 16.5, 14.0, 13.7, 13.1, 12.5 (2C), 12.3
(2C, CH, CH3). Anal. Calcd for C55H82FN3O12SSi2 (1084.49): C,
60.91; H, 7.62; N, 3.87; S, 2.96. Found: C, 60.75; H, 7.56; N, 3.89;
S, 2.88.
lowish oil. [
a
]
D ꢀ18.3 (c 0.9, CHCl3). 1H NMR (CDCl3): d 8.70 (s,1H,
NH), 8.08–7.39 (m, 5H, PhH), 6.37 (d, 1H, J1,2 = 1.7 Hz, H-1), 5.72
(dd, 1H, J2,3 = 3.4 Hz, H-2), 5.52 (t, 1H, J3,4 = J4,5 = 9.8 Hz, H-4),
5.39 (dd, 1H, H-3), 4.18 (dd, 1H, J5,6a = 2.1Hz, J6a,6b = ꢀ12.7 Hz, H-
6a), 3.97 (dd, 1H, J5,6b = 1.1Hz, H-6b), 3.88 (br d, J = 9.5 Hz, H-
5),1.98 (s, 3H, Acetyl-CH3), 1.14–0.96 (m, 28H, CH, CH3). 13C NMR
(CDCl3): d 170.2 (C@NH), 165.4, 160.1 (C@O), 133.6, 130.1, 129.2,
128.5 (PhC), 95.8 (C-1), 90.6 (CCl3), 76.3 (C-5), 71.8 (C-3), 68.9
(C-2), 64.1 (C-4), 60.7 (C-6), 21.1 (CH3), 17.6, 17.4 (2C), 17.3 (2C),
17.2, 13.6, 13.3, 12.6 (2C, CH, CH3). Anal. Calcd for C29H44Cl3O9Si2
(713.19): C, 48.84; H, 6.22; N, 1.96. Found: C, 48.86; H, 6.16; N,
1.91.

2356
R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
3.13. Octyl 2,3,4,6-tetra-O-benzoyl-
a
-
D
-mannopyranosyl-(1?6)-
(C-10), 101.6 (C-1), 84.2 (C-1Glc), 82.8 (C-3Glc), 77.5 (C-4Glc), 76.1
(PhCH2), 74.4 (PhCH2), 74.4, 74.3, 72.6 (2C), 72.4, 72.2, 72.0 (C-
3-O-acetyl-2-O-benzoyl-4-O-(1-fluoro-1,1,3,3-tetraisopropyl-
1,3-disiloxane-3-yl)- -mannopyranosyl-(1?4)-2-azido-3,6-
di-O-benzyl-2-deoxy-1-thio- -glucopyranoside (16)
a-D
5
Glc, C-20, C-30, C-50, C-200, C-300, C-400), 70.9 (C-6Glc), 68.6 (C-500),
a-
D
68.2 (C-40), 67.4 (C-60), 65.2 (C-2Glc), 62.8 (C-600), 33.0, 31.1, 30.6,
30.3, 30.2, 30.0, 23.7 (CH2), 14.5 (CH3). HRMS ESI: Calcd for
A solution of 1543 (237 mg, 0.36 mmol) and sym-collidine
(35 mL, 0.26 mmol) in dichloromethane (15 mL) was slowly added
at ꢀ25 °C to a stirred slurry of 14 (339 mg, 0.33 mmol), AgOTf
(92 g, 0.36 mmol) and 4 Å molecular sieves (0.5 g) in dichloro-
methane (15 mL) and stirring was continued for 1 h. Et3N
(1.0 mL) was added and the slurry was filtered through a layer of
Celite. The filtrate was successively washed with aqueous Na2S2O3,
HCl and Na2CO3 solutions, dried, filtered and concentrated. Chro-
matography of the residue with 50/1 toluene-ethyl acetate gave
crude 16 which was directly used for the next step.
C
40H59N3NaO14S (M+Na+): m/z 860.3616; Found: m/z 860.3618.
3.16. Octyl
a-
D
-mannopyranosyl-(1?6)-
a-D-mannopyranosyl-
(1?4)-2-amino-2-deoxy-1-thio-
a-D-glucopyranoside (19)
Sodium (75 mg, 0.11 mmol) was added in three portions within
10 minutes at ꢀ78 °C to a stirred solution of 18 (90 mg, 0.11 mmol)
in THF (5 mL) and liquid NH3 (15 mL). Stirring was continued for
15 minutes, NH4Cl (0.25 g) was added to the mixture in one por-
tion and the solvent was removed at rt in a stream of Ar. Successive
chromatography of the residue with 10/2/1/3 n-butanol–ethanol–
ammonia–water on silica gel and with water on Biogel P2, followed
by filtration through a Sulpelco RP C18 Superclean cartridge with a
1/1 water–methanol mixture and lyophyilization gave pure 19
3.14. Octyl 2,3,4,6-tetra-O-benzoyl-
3-O-acetyl-2-O-benzoyl- -mannopyranosyl-(1?4)-2-azido-
3,6-di-O-benzyl-2-deoxy-1-thio- -glucopyranoside (17)
a-D-mannopyranosyl-(1?6)-
a-D
a
-D
(34 mg, 51%) as a white foam. [a]
+119 (c 0.1, MeOH). 1H NMR
D
A solution of crude 16 and Bu4NF trihydrate (100 mg) in THF
was stirred at rt for 1 h and worked up as described for the prepa-
ration of compound. 14. Chromatography with 10/1 toluene–ethyl
(MeOD-d4): d 5.26 (d, 1H, J1Glc,2Glc = 5.3 Hz, H-1Glc), 5.25 (d, 1H,
J1,2 = 1.8 Hz, H-1), 4.82 (H-10), 4.00 (ddd, 1H, J4Glc,5Glc = 9.4 Hz,
J5Glc,6aGlc = 4.0 Hz, J5Glc,6bGlc = 3.0 Hz, H-5Glc), 3.95 (dd, 1H,
J2,3 = 2.8 Hz, H-2), 3.89 (H-6a), 3.87 (H-20), 3.80 (H-6a0, H-6aGlc, H-
6bGlc), 3.78 (H-30), 3.77 (H-5), 3.75 (H-6b), 3.71 (H-6b0), 3.67 (H-
3), 3.66 (H-50), 3.65 (H-4), 3.62 (H-40), 3.52 (t, 1H, J3Glc,4Glc = 8.8 Hz,
H-4Glc), 3.44 (dd, 1H, J2Glc,3Glc = 9.8 Hz, H-3Glc), 2.90 (dd, 1H, H-2Glc),
2.69–2.69 (m, 2H, CH2), 1.60–1.21 (m, 12H, CH2), 0.89 (t, 3H,
acetate gave 17 (353 mg, 76% in rel. to 14) as colorless oil. [
a]
D
+32.5 (c 1.1, CHCl3). 1H NMR (CDCl3): d 8.13–6.96 (m, 35H, PhH),
6.14 (t, 1H, J3 ,4 = J4 ,5 = 10.0 Hz, H-400), 5.90 (dd, 1H, J2 ,3 = 3.2 Hz,
00 00
00 00
00 00
H-300), 5.82 (dd, 1H, J1 ,2 = 1.7 Hz, H-200), 5.61 (dd, 1H, J1 ,2 = 2.9 Hz,
H-20), 5.50 (d, 1H, H-10), 5.44 (d, 1H, J1,2 = 5.3 Hz, H-1), 5.23 (dd,
00 00
0
0
13
J3 ,4 = 5.2 Hz, H-30), 5.08 (d, 1H, H-100), 4.94 (d, 1H, J = ꢀ10.6 Hz,
PhCH2), 4.77 (d, 1H, J = ꢀ10.7 Hz, PhCH2), 4.71–4.63 (m, 1H, H-
6a00), 4.61 (s, 2H, PhCH2), 4.46–4.41 (m, 2H, H-6b00, H-500), 4.31–
4.22 (m, 2H, H-40, H-5), 4.03–3.60 (m, 9H, H-2, H-3, H-40, H-50, H-
500, H-6, H-600), 2.71–2.55 (m, 2H, SCH2), 2.49 (d, 1H, J = 4.9 Hz,
OH), 2.08 (s, 3H, Acetyl-CH3), 1.68–1.61 (m, 2H, CH2), 1.39–1.03
(m, 10H, CH2), 0.90–0.83 (m, 3H, CH3). 13C NMR (CDCl3): d 171.2,
166.2, 165.3 (2C), 165.2, 164.9 (C@O), 137.8, 136.9, 133.4, 133.3,
133.2, 133.1, 129.8, 19.7, 129.6 (2C), 129.1, 129.0 (2C), 128.9,
128.6 (2C), 128.5, 128.3, 128.2 (2C), 127.7, 127.6 (3C), 127.4,
J = 6.2 Hz, CH3). C NMR (MeOD-d4): d 102.8 (C-1), 101.5 (C-10),
0
0
88.0 (C-1Glc), 78.7 (C-4Glc), 76.6 (C-3Glc), 74.4 (C-50), 74.1 (C-5),
73.3 (C-5Glc), 72.5 (2C, C-3, C-30), 72.2 (C-2), 72.0 (C-20), 68.6 (C-
40), 68.5 (C-4), 67.9 (C-6), 62.9 (C-60), 62.7 (C-6Glc), 57.4 (C-2Glc),
33.0, 32.1, 30.9, 30.4, 30.3, 30.0, 23.7 (CH2), 14.4 (CH3). HRMS
ESI: Calcd for C26H49NO14S (M+H+): m/z 632.295; Found: m/z
632.293.
3.17. Ethyl 2,3,4,6-tetra-O-benzyl-1-sulfinyl-b-D-
galactopyransoside (20b)
125.2 (PhC), 98.9 (JC1 ,H1 = 174.7 Hz, C-10), 97.9 (JC1 ,H1 = 175.3 Hz,
C-100), 83.2 (JC1,H1 = 166.4 Hz, C-1), 81.6 (C-3), 75.2 (PhCH2), 75.1
(C-4), 73.2 (PhCH2), 72.4 (2C, C-50, C-30), 70.4 (C-5), 70.0 (C-300),
69.8 (2C, C-20, C-200), 68.7 (C-6), 68.5 (C-500), 66.5 (C-400), 65.8 (C-
60), 65.1 (C-40), 64.2 (C-2), 62.6 (C-600), 31.8, 30.5, 29.5, 29.1 (2C),
28.9, 22.6 (CH2), 21.0 (Acetyl-CH3), 14.1 (CH3). Anal. Calcd for
0
0
00
00
m-Chloroperbenzoic acid (50%, 967 mg, 2.8 mmol) was added at
ꢀ25 °C to a stirred solution of 20a30 (1.50 g, 2.55 mmol) in dichlo-
romethane (110 mL). The mixture is warmed to rt, washed with
aqueous NaHCO3 solution, dried, filtered and concentrated. Chro-
matography of the residue with 5/1 toluene–ethyl acetate gave
C
77H81N3O20S (1400.54): C, 66.03; H, 5.83; N, 3.00; S, 2.29. Found:
20b (1.52 g, 99%) as a white amorphous solid. [
a
]
ꢀ40.9 (c 1,
D
C, 66.36; H, 5.94; N, 2.88; S, 2.33.
CHCl3). 13C-NMR (CDCl3): d 138.4, 138.0, 137.9, 137.7, 128.6,
128.5, 128.4 (2C), 128.2, 128.1, 127.9, 127.8, 127.5, 127.4 (PhC),
89.6 (C-1), 84.1 (C-5), 78.7 (C-2), 75.9 (PhCH2), 74.4 (PhCH2), 73.6
(PhCH2), 73.0 (2C, C-3, C-4), 72.5 (PhCH2), 68.9 (C-6), 40.7 (SCH2),
7.4 (CH3). Anal. Calcd for C36H40O6S (600.77): C, 71.97; H, 6.71; S,
5.34. Found: C, 71.92; H, 6.69, S, 5.43.
3.15. Octyl a-D-mannopyranosyl-(1?6)-a-D-mannopyranosyl-
(1?4)-2-azido-3,6-di-O-benzyl-2-deoxy-1-thio-a-D-
glucopyranoside (18)
A solution of 17 (336 mg, 0.24 mmol) and 1 M NaOMe in MeOH
(ca. 1 L) in MeOH (10 mL) was stirred at rt for 16 h. Work up as
described for the preparation of compound 5 and lyophilization
of the residue gave 18 (188 mg, 93%) as white foam. [ +119.4
(c 0.8, MeOH). 1H NMR (MeOD-d4): d 7.44–7.25 (m, 10H, PhH),
l
3.18. Octyl 2,3,4,6-tetra-O-benzyl-
2-O-benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane-1,3-
diyl)-1-thio- -mannopyranoside (21)
a-D-galactopyranosyl-(1?3)-
a]
a-D
D
0
0
5.46 (d, 1H, J1Glc,2Glc = 5.4 Hz, H-1Glc), 5.24 (d, 1H, J1 ,2 = 1.62 Hz,
(a) Bromine (12
stirred slurry of
0.36 mmol), 4 Å molecular sieves (0.5 g), AgOTf (154 mg,
0.60 mmol) and tetramethyl urea (72 L, 0.60 mmol) in toluene
(6 mL), and stirring was continued for 2 h while the mixture was
gradually allowed to warm to rt. Et3N (0.12 mL) was added, the
mixture filtered through a layer of Celite, and the filtrate washed
successively with aqueous Na2S2O3 and NaHCO3 solutions, dried
and concentrated. Chromatography of the residue with 1/3 n-hex-
ane–dichloromethane gave first 21 (156 mg, 46%) as a white
l
L, 0.23 mmol) was added with syringe at 0 °C
H-10), 4.91–4.78 (m, 18H, Bn, OH), 4.76 (d, 1H, J1 ,2 = 1.62 Hz, H-
to
a
9
(188 mg, 0.29 mmol), 20a (212 mg,
00 00
100), 4.73 (d, 1H, J = ꢀ10.6 Hz, PhCH2), 4.61 (d, 1H, J = 11.8 Hz,
PhCH2), 4.55 (d, 1H, J = 11.8 Hz, PhCH2), 4.19 (dt, 1H,
J4Glc,5Glc = 9.2 Hz, J5Glc,6aGlc = J5Glc,6bGlc = 3.8 Hz, H-5Glc), 3.96 (dd,
1H, J2Glc,3Glc = 9.8 Hz, H-2), 3.90–3.59 (m, 16H, H-3Glc, H-4Glc, H-
6a,bGlc, H-20, H-30, H-40, H-50, H-6a,b0, H-200, H-300, H-400, H-500, H-
6a,b00), 2.67–2.53 (m, 2H, SCH2), 1.65–1.57 (m, 2H, CH2), 1.37–
1.28 (m, 10H, CH2), 0.89 (d, 1H, J = 6.7 Hz, CH3). 13C NMR (MeOD-
d4): d 139.7, 139.0, 129.4 (3C), 128.9, 128.8, 128.6 (PhC), 103.3
l

R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2357
amorphous solid. [
a
]
D +25.9 (c 0.9, CHCl3). 1H NMR (CDCl3): d8.05–
(1.72 mL, 12.9 mmol) and 4 Å molecular sieves (2.0 g) in dichloro-
methane (50 mL), and stirring was continued for 1.5 h while the
mixture was allowed to warm up to 5 °C. Workup as described un-
der (a) and chromatography of the residue with 10:1 n-hexane–
ethyl acetate gave 21 (0.78 g, 61%).
6.98 (m, 25H, PhH), 5.51 (br s, 2H, H-1, H-2), 4.91 (d, 1H, J1Gal,2-
Gal = 3.5 Hz, H-1Gal), 4.82 (d, 1H, J = ꢀ11.3 Hz, PhCH2), 4.77 (d, 1H,
J = ꢀ12.6 Hz, PhCH2), 4.75 (d, 1H, J = ꢀ11.6 Hz, PhCH2), 4.66 (d,
1H, J = ꢀ11.6 Hz, PhCH2), 4.65 (t, 1H, J3,4 = J4,5 = 9.1 Hz, H-4), 4.63
(d, 1H, J = ꢀ12.5 Hz, PhCH2), 4.34 (dd, 1H, J5,6a = 1.5 Hz,
J6a,6b = ꢀ12.6 Hz, H-6a), 4.03–3.82 (m, 10H, H-3, H-5, H-6b, H-
3.19. 2,3,4,6-Tetra-O-benzyl-
benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane-1,3-diyl)-
-mannopyranose (22)
a-D-galactopyranosyl-(1?3)-2-O-
2Gal, H-3Gal, H-4Gal, H-5Gal, PhCH2), 3.50 (t, 1H, J = 8.9 Hz, H-6aGal),
3.16 (dd, 1H, J5Gal,6bGal = 4.7 Hz, J6aGal,6bGal = 8.4 Hz, H-6bGal), 2.67–
2.54 (m, 2H, SCH2), 1.64–1.59 (m, 2H, CH2), 1.37–0.91 (m, 41H,
CH, CH2, CH3), 0.90–0.86 (m, 3H, CH3). 13C NMR (CDCl3): d 165.9
(C@O), 139.0, 138.9, 137.9, 133.2, 129.9, 129.7, 129.0, 128.4,
128.3, 128.2, 128.1 (2C), 127.9, 127.8, 127.7, 127.5, 127.4, 127.3
(2C), 127.2 (PhC), 100.9 (C-1Gal), 82.8 (C-1), 81.0 (C-3), 78.9 (C-
a-D
Treatment of compound. 21 (1.68 g, 1.43 mmol) with NBS
(0.41 g, 2.32 mmol) in aqueous acetone (1% water content,
50 mL) as described for the preparation of compound. 11 and chro-
matography with 15:1 toluene–ethyl acetate gave crude 22 (0.96 g,
64%) as a colorless oil which was directly used for the next step.
3Gal), 76.6 (C-2Gal), 75.3 (C-5), 74.8 (PhCH2), 74.7 (C-4Gal), 74.4 (C-
2), 73.2 (PhCH2), 73.1 (PhCH2), 72.9 (PhCH2), 69.9 (C-5Gal), 67.7
(C-6Gal), 66.2 (C-4), 61.1 (C-6), 31.8 (2C), 29.8, 29.2, 29.1, 28.8,
22.7 (CH2), 17.9, 17.8, 17.6, 17.4 (3C), 17.3, 17.1, 14.1, 13.3, 13.0,
12.9, 12.7 (CH, CH3). Anal. Calcd for C87H92O12SSi2 (1177.683): C,
68.33; H, 7.86. Found: C, 68.12; H, 7.72.
3.20. 2,3,4,6-Tetra-O-benzyl-
benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane-1,3-diyl)-
-mannopyranosyl trichloroacetimidat (23)
a-D-galactopyranosyl-(1?3)-2-O-
a-D
Eluted next was not consumed starting material 9 (25 mg, 13%).
(b) Performing the reaction as described under (a) but in dichlo-
romethane instead of toluene resulted in a complex inseperable
mixture of unidentified products.
Treatment of crude 22 (0.91 g, 0.87 mmol) with trichloroaceto-
nitrile (0.92 ml) as described for the preparation of compound 12
and chromatography with 50:1 toluene–ethyl acetate gave 23
(0.63 g, 72%) as a colorless oil. [
a]
D
ꢀ12.0 (c 0.1, CHCl3). 1H NMR
(c) MeOTf (114
lL, 1.05 mmol) was added at rt to a stirred slur-
(CDCl3): d 8.72 (s, 1H, C@NH), 8.07–7.00 (m, 25H, PhH), 6.53 (d,
1H, J1,2 = 1.5 Hz, H-1), 5.55 (br t, 1H, J = 2.4 Hz, H-2), 4.86 (d, 1H,
J1Gal,2Gal = 3.7 Hz, H-1Gal), 4.83 (d, 1H, J = 11.5 Hz, PhCH2), 4.78 (d,
1H, J = ꢀ11.6 Hz, PhCH2), 7.76 (d, 1H, J = 11.8 Hz, PhCH2), 4.74 (t,
1H, J3,4 = J4,5 = 9.6 Hz, H-4), 4.71 (d, 1H, J = ꢀ12.9 Hz, PhCH2), 4.67
(d, 1H, J = ꢀ11.6 Hz, PhCH2), 4.59 (d, 1H, J = ꢀ12.6 Hz, PhCH2),
4.43 (d, 1H, J = ꢀ11.3 Hz, PhCH2), 4.20 (dd, 1H, J5,6a = 1.8 Hz,
J6a,6b = ꢀ12.6 Hz, H-6a), 4.14 (dd, 1H, H-3), 4.06–3.85 (m, 5H, H-
ry of 9 (328 mg, 0.50 mmol), 20a (585 mg, 1.0 mmol), 2,6-di-t-bu-
tyl pyridine (0.62 mL, 2.8 mmol) and 4 Å molecular sieves (1.0 g) in
diethylether (10 mL) and the mixture was stirred at rt for 24 h.
After addition of Et3N (1.0 mL) and dichloromethane (50 mL) the
mixture was worked up as described under (a). Chromatography
of the residue with 50/1 toluene-ethyl acetate gave first 21
(108 mg, 18%). Eluted next was not consumed starting material 9
(136 mg, 42%).
2
Gal, H-3Gal, H-4Gal, H-5Gal, PhCH2), 3.79 (br d, J = 9.4 Hz, H-6b),
(d) Performing the reaction as described under (a) but with N-
iodosuccinimide and TMSOTf in dichloromethane instead of bro-
mine in toluene resulted in a complex inseperable mixture of
unidentified products.
(e) Performing the reaction as described under (a) but with
iodoniumdicollidine perchlorate in dichloromethane instead of
bromine in toluene resulted in a complex inseperable mixture of
unidentified products.
3.50 (br t, 1H, J = 8.8 Hz, H-6aGal), 3.19 (dd, 1H, J5Gal,6bGal = 4.7 Hz,
J6aGal,6bGal = 8.4 Hz, H-6bGal), 1.28–0.98 (m, 28H, CH, CH3). 13C
NMR (CDCl3): d 165.7 (C@O), 159.7 (C@NH), 139.1, 139.0, 138.8,
137.8, 133.4, 129.8, 129.4, 129.0, 128.4, 128.3, 128.2, 128.1 (2C),
127.9, 127.8 (2C), 127.5, 127.3 (2C), 127.1 (PhC), 101.5 (C-1Gal),
94.7 (C-1), 80.2 (C-3), 78.9 (C-3Gal), 76.7 (2C, C-5, C-2Gal), 74.8
(PhCH2), 74.6 (C-4Gal), 73.5 (PhCH2), 73.2 (PhCH2), 73.0 (PhCH2),
71.9 (C-2), 69.9 (C-5Gal), 67.5 (C-6Gal), 65.5 (C-4), 60.6 (C-6Gal),
17.8, 17.7, 17.5, 17.4 (3C), 17.3, 17.0, 13.2, 13.0 (2C), 12.8 (CH,
CH2). Anal. Calcd for C61H76Cl3O13Si2 (1193.78): C, 61.37; H, 6.42;
N, 1.17. Found: C, 61.34; H, 6.44; N, 1.17.
(f) Tf2O (131
solution of 9 (328 mg, 0.50 mmol), 20b (300 mg, 0.5 mmol), sym-
collidine (67 L, 0.5 mmol) in a 1:1 mixture of toluene and dichlo-
l
L, 0.5 mmol) was added at ꢀ50 °C to a stirred
l
romethane (8 mL) and the mixture was stirred at for 30 min. TLC
showed the formation of a complex inseperable mixture of uniden-
tified products.
3.21. Octyl 2,3,4,6-tetra-O-benzyl-
2-O-benzoyl-4,6-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane-1,3-
diyl)- -mannopyranosyl-(1?4)-2-azido-3,6-di-O-benzyl-2-
deoxy-1-thio-
a-D-galactopyranosyl-(1?3)-
(g) TMSOTf (10
l
L) was added at ꢀ30 °C to a stirred solution of
a-D
9 (314 mg, 0.48 mmol) and 20c31 (411 mg, 0.60 mmol) in 1:1
dichloromethane and diethylether (7 mL) and stirring was contin-
ued for 18 h. TLC showed the formation of a complex inseperable
mixture of unidentified products.
a-
D
-glucopyranoside (24)
TMSOTf (18
l
L) was added at ꢀ20 °C to a stirred slurry of 23
(667 mg, 0.56 mmol), 8 (430 mg, 0.84 mmol) and 4 Å molecular
sieves (2 g) in diethylether (50 mL) and the mixture was stirred
for 1.5 h. After neutralization of the mixture by addition of solid
NaHCO3, the mixture was filtered through a layer of Celite and
the filtrate was washed with aqueous NaHCO3 solution, dried
and concentrated. Chromatography of the residue with 40:1 tolu-
(h) A solution of 2,3,4,6-tetra-O-benzyl-a-D-galactopyranosyl
chloride (20d)32 (1.74 g, 3.13 mmol) in dichloromethane (30 mL)
was added within 1 h at 0 °C to a stirred slurry of 9 (1.57 g,
2.5 mmol), AgClO4 (1.24 g, 5.98 mmol), sym-collidine (0.75 mL,
5.62 mmol) and 4 Å molecular sieves (3.0 g) in dichloromethane
(20 mL), and stirring was continued for 2 h while the mixture
was allowed to warm up to rt. Workup as described under (a)
and chromatography of the residue with 10:1 n-hexane–ethyl ace-
tate gave 21 (1.51 g, 51%).
ene–ethyl acetate gave 24 (789 mg, 91%) as a colorless oil. [a]
D
+32.6 (c 1.7, CHCl3). 1H NMR (CDCl3): d 7.95–6.97 (m, 35H, PhH),
5.53 (dd, 1H, J1,2 = 1.6 Hz, J2,3 = 2.8 Hz, H-2), 5.50 (d, 1H, H-1),
5.40 (d, 1H, J1Glc,2Glc = 5.3 Hz, H-1Glc), 4.97 (d, 1H, J = ꢀ10.9 Hz,
PhCH2), 4.93 (d, 1H, J1Gal,2Gal = 3.2 Hz, H-1Gal), 4.81 (d, 1H,
J = ꢀ11.3 Hz, PhCH2), 4.78 (d, 1H, J = ꢀ12.3 Hz, PhCH2), 4.75 (d,
1H, J = ꢀ10.6 Hz, PhCH2), 4.73 (d, 1H, J = ꢀ11.6 Hz, PhCH2), 4.66–
(i) A solution of 20d (0.92 g, 1.65 mmol) in dichloromethane
(20 mL) was added within 30 min at ꢀ5 °C to a stirred slurry of 9
(0.7 g, 1.12 mmol), AgClO4 (1.50 g, 7.24 mmol), sym-collidine

2358
R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
4.53 (m, 5H, PhCH2, H-4), 4.43 (d, 1H, J = ꢀ11.3 Hz, PhCH2), 4.28–
4.25 (br m, 1H, H-6a), 4.05–3.70 (m, 14H, H-3, H-6b, H-2Glc, H-
3Glc H-4Glc, H-5Glc, H-6Glc, H-2Gal, H-3Gal, H-4Gal, H-5Gal, PhCH2),
3.56 (br d, 1H, J = 11.3 Hz, H-5), 3.46 (t, 1H, J = 8.9 Hz, H-6aGal),
3.13 (dd, 1H, J6aGal,6bGal = ꢀ8.5 Hz, J5Gal,6bGal = 4.6 Hz, H-6bGal),
2.69–2.53 (m, 2H, SCH2), 1.70–1.60 (m, 2H, CH2), 1.38–0.82 (m,
41H, CH, CH2, CH3). 13C NMR (CDCl3): d 165.5 (C@O), 139.2,
139.0, 138.9, 138.0, 137.9, 133.0, 129.7 (2C), 129.0, 128.3, 128.2
(2C), 128.1 (2C), 128.0 (2C), 127.8 (2C), 127.6, 127.5 (2C), 127.4,
127.3, 127.3, 127.2, 127.1 (2C), 125.3 (PhC), 100.8 (JC1Gal,H1-
Gal = 167.0 Hz, C-1Gal), 98.6 (JC1,H1 = 170.3 Hz, C-1), 83.3
(JC1Glc,H1Glc = 166.4 Hz, C-1Glc), 81.4 (C-3Glc), 79.8 (C-3), 78.9 (C-
138.7, 138.4, 138.2, 138.1, 137.7, 137.3, 133.3, 133.1, 132.9,
130.0, 129.8, 129.7, 129.6, 129.4, 129.2, 129.0, 128.5 (2C), 128.4,
128.3 (2C), 128.2, 128.1, 128.0 (2C), 127.9, 127.7, 127.6, 127.5,
127.4, 127.3, 102.3 (C-1Gal), 99.2 (C-1), 97.9 (C-10), 83.2 (C-1Glc),
81.5 (C-3Glc), 81.5 (C-3), 79.6 (C-3Gal), 76.3 (C-2Gal), 75.4 (C-4Glc),
75.1 (PhCH2), 74.8 (PhCH2), 74.5 (PhCH2), 74.2 (C-4Gal), 73.5
(PhCH2), 73.1 (PhCH2), 72.6 (C-5), 72.3 (PhCH2), 71.8 (C-2), 70.8
(C-5Glc), 70.3 (C-30), 70.2 (C-20), 69.6 (C-5Gal), 69.2 (C-6Glc), 68.7
(C-50), 67.8 (C-6Gal), 66.7 (C-40), 66.6 (C-4), 66.5 (C-6), 64.2 (C-
2
Glc), 62.6 (C-60), 31.8, 30.5, 29.5, 29.2 (2C), 29.0, 22.6 (CH2), 14.1
(CH3). FAB-MS (pos.): m/z 1903.6 [M+Na]+. Anal. Calcd for
109H113N3O24S (1881.14): C, 69.59; H, 6.05; N, 2.23. Found: C,
69.39; H, 6.16; N, 2.16.
(b) TMSOTf (5
L) was added at ꢀ20 °C to a stirred slurry of
compound 31 (280 mg, 0.16 mmol), compound (120 mg,
C
3
Gal), 76.7 (C-2Gal), 75.9 (C-4Glc), 75.2 (PhCH2), 74.8 (PhCH2), 74.7
(C-4Gal), 73.3 (PhCH2), 73.3 (3C, C-5⁄, C-2⁄, PhCH2), 73.1 (PhCH2),
72.8 (PhCH2), 70.8 (C-5Glc), 69.2 (C-5Gal), 69.2 (C-6Glc), 67.7 (C-
l
8
6
Gal), 65.8 (C-4), 64.5 (C-2Glc), 60.7 (C-6), 31.8, 29.5, 29.1 (2C),
0.23 mmol) and 4 Å molecular sieves (0.5 g) in diethylether
(20 mL) and stirring was continued for 1.5 h. Workup as described
for the preparation of compound 13 followed by treatment of the
intermediate with Bu4NF trihydrate (50 mg) in THF (10 mL) as de-
scribed for the preparation of 17 and chromatography with 30:1
toluene–ethyl acetate gave 26 (128 mg, 44%).
29.0, 22.6 (CH2), 17.8, 17.7, 17.4 (3C), 17.3, 17.1, 14.1 (CH3), 13.2,
12.9 (2C), 12.7 (CH). Anal. Calcd for C87H113N3O16SSi2 (1545.08):
C, 7.63; H, 7.37; N, 2.72; S, 2.1. Found: C, 67.55; H, 7.27; N, 2.55;
S, 2.1.
3.22. Octyl 2,3,4,6-tetra-O-benzyl-
2-O-benzoyl-4-O-(1-fluoro-1,1,3,3-tetraisopropyl-1,3-disil-
oxane-3-yl)- -mannopyranosyl-(1?4)-2-azido-3,6-di-O-
benzyl-2-deoxy-1-thio- -glucopyranoside (25)
a-D-galactopyranosyl-(1?3)-
3.24. Octyl
(1?3)]-
a-D-glucopyranoside (27)
a
-
D
-mannopyranosyl-(1?6)-[
a-D-galactopyranosyl-
a
-D
a-D-mannopyranosyl-(1?4)-2-amino-2-deoxy-1-thio-
a-
D
Treatment of compound 24 (300 mg, 0.19 mmol) with HF in
pyridine (70%, 38 L) in dichloromethane (15 mL) as described
for the preparation of 14 and chromatography with 20:1 tolu-
ene–ethyl acetate gave 25 (267 mg, 89%) as a colorless oil which
was used for the next step without further characterization. [a]
D
Compound 26 (105 mg, 0.06 mmol) was deprotected with a cat-
l
alytic amount of NaOMe in MeOH as described for the preparation
of compound 18 followed by treatment of the intermediate with
Na (71 mg, 3.09 mmol) in THF (5 mL) and liquid NH3 (15 mL) as de-
scribed for the preparation of compound 19. Purification as de-
scribed for the preparation of compound 19 gave pure 27 (29 mg,
+55.0 (c 0.9, CHCl3). Anal. Calcd for C87H114FN3O16SSi2 (1565.08):
C, 66.77; H, 7.34; N, 2.68. Found: C, 66.76; H, 7.40; N, 2.65.
64%). [
a]
+222 (c 0.1, MeOH). 1H NMR (MeOD-d4): d 5.37 (d, 1H,
D
J1Glc,2Glc = 5.0 Hz, H-1Glc), 5.22 (d, 1H, J1,2 = 1.6 Hz, H-1), 5.15 (d,
0
0
3.23. Octyl 2,3,4,6-tetra-O-benzoyl-
[2,3,4,6-tetra-O-benzyl- -galactopyranosyl-(1?3)]-2-O-ben-
zoyl- -mannopyranosyl-(1?4)-2-azido-3,6-di-O-benzyl-2-
deoxy-1-thio- -glucopyranoside (26)
a-D-mannopyranosyl-(1?6)-
1H, J1Gal,2Gal = 3.2 Hz, H-1Gal), 4.83 (d, 1H, J1 ,2 = 1.4 Hz, H-10), 4.25
(br s, 1H, H-2), 4.10 (dd, 1H, J = 7.7 Hz, J = 4.0 Hz, H-5Gal), 4.02
(dt, J4Glc,5Glc = 9.0 Hz, J5Glc, 6aGlc = J5Glc,6bGlc = 3.2 Hz, H-5Glc), 3.91
(H-6a), 3.87 (2H, H-20, H-4Gal), 3.86 (H-4), 3.83 (H-5), 3.82 (2H,
H-6aGlc, H-6bGlc), 3.81 (2H, H-6a0, H-3Gal), 3.79 (H-6aGal), 3.78
(3H, H-30, H-3, H-2Gal), 3.76 (H-6b), 3.75 (H-6b0), 3.70 (H-6bGal),
3.65 (3H, H-40, H-50, H-3Glc), 3.57 (t, 1H, J3Glc,4Glc = 9.2 Hz, H-4Glc),
3.20 (dd, 1H, J2glc,3Glc = 9.8 Hz, H-2Glc), 2.67 (t, 2H, J = 7.2 Hz,
SCH2), 1.68–1.63 (m, 2H, CH2), 1.40–1.28 (m, 10H, CH2), 0.89 (t,
3H, J = 6.9 Hz, CH3). 13C NMR (MeOD-d4): d 103.2 (C-1), 102.3 (C-
a-D
a-D
a
-D
(a) Treatment of 25 (191 mg, 0.12 mmol), AgOTf (34 mg,
0.13 mmol), 4 Å molecular sieves (1 g) in dichloromethane
(10 mL) with 15 (89 mg, 0.13 mmol) and sym-collidine (13 lL,
0.1 mmol) in dichloromethane (5 mL) at ꢀ25 °C as described for
the preparation of 16, followed by treatment of the intermediate
with Bu4NF trihydrate (50 mg) in THF (10 mL) as described for
the preparation of 17 and chromatography with 30:1 toluene–
1
Gal), 102.0 (C-10), 86.0 (C-1Glc), 81.5 (C-3), 78.7 (C-4Glc), 74.4 (C-
50), 74,2 (C-5), 74.1 (C-3Glc), 73.5 (C-5Glc), 73.0 (C-5Gal), 72.5 (C-
30), 72.0 (C-20), 71.5 (C-3Gal), 71.3 (2C, C-2, C-4Gal), 70.8 (C-2Gal),
68.6 (C-40), 67.7 (C-6), 67.2 (C-4), 63.2 (C-6Gal), 62.8 (C-60), 62.3
(C-6Glc), 56.5 (C-2Glc), 33.0, 32.6, 30.9, 30.4, 30.3, 29.9, 23.7 (CH2),
14.4 (CH3). HRMS ESI Calcd for (C32H60NO19S) (M+H+): m/z
794.348; Found: m/z 794.348.
ethyl acetate gave 26 (157 mg, 68%). [
a]
+42.0 (c 0.6, CHCl3). 1H
D
NMR (CDCl3): 8.13–7.02 (m, 55H, PhH), 6.13 (t, 1H,
d
J3 ,4 = J4 ,5 = 10 Hz, H-40), 5.94 (dd, 1H, J2 ,3 = 3.2 Hz, H-30), 5.78
0
0
0
0
0
0
(dd, 1H, J1 ,2 = 1.5 Hz, H-20), 5.64 (dd, 1H, J1,2 = 1.5 Hz, J2,3 = 2.9 Hz,
H-2), 5.47 (d, 1H, J1,2 = 1.2 Hz, H-1), 5.42 (d, 1H, J1Glc,2Glc = 5.3 Hz,
H-1Glc), 5.08 (d, 1H, H-10), 5.01 (d, 1H, J1Gal,2Gal = 3.7 Hz, H-1Gal),
4.88 (d, 1H, J = ꢀ11.6 Hz, PhCH2), 4.82 (d, 1H, J = ꢀ11.2 Hz, PhCH2),
4.81 (br s, 2H, PhCH2), 4.73 (d, 1H, J = ꢀ11.6 Hz, PhCH2), 4.65 (H-
6a0), 4.65 (2H, PhCH2), 4.62 (2H, PhCH2), 4.51 (d, 1H, J = ꢀ11.3 Hz,
PhCH2), 4.44 (H-50), 4.39 (2H, H-6b0, PhCH2-4a), 4.34 (d, 1H,
J = ꢀ11.8 Hz, PhCH2), 4.29 (H-5Glc), 4.18 (t, 1H, J3,4 = J4,5 = 9.4 Hz,
H-4), 4.07 (dd, 1H, J2Gal,3Gal = 9.4 Hz, H-2Gal), 4.00 (2H, H-6a, H-
0
0
3.25. Octyl 2,3,4,6-Tetra-O-benzyl-
2-O-benzoyl-4-O-(3-fluoro-1,1,3,3-tetraisopropyl-1,3-
disiloxane-1-yl)-1-thio- -mannopyranoside (28)
a-D-galactopyranosyl-(1?3)-
a
-D
Treatment of compound 21 (565 mg, 0.48 mmol) with HF in
pyridine (70%, 50 L) in dichloromethane (20 mL) as described
for the preparation of 14 and chromatography with 70:1 tolu-
l
4Glc), 3.98 (H-5Gal), 3.96 (H-6aGlc), 3.95 (H-5), 3.93 (H-4Gal), 3.92
(H-2Glc), 3.91 (2H, H-3, H-3Gal), 3.84 (t, 1H, J = 9.5 Hz, H-6bGlc),
3.79 (dd, 1H, J2,3 = 8.9 Hz, J3,4 = 9.8 Hz, H-3Glc), 3.72 (br s, 1H, H-
6b), 3.55 (t, 1H, J = 8.7 Hz, H-6aGal), 3.44 (dd, 1H, J5Gal,6bGal, H-6bGal),
2.63 (m, 2H, SCH2), 1.66–1.25 (m, 12H, CH2), 0.87 (br t, J = 6.7 Hz,
CH3). 13C NMR (CDCl3): d 166.2, 165.4, 165.3 (2C), 165.0 (C@O),
ene–ethyl acetate gave 28 (544 mg, 95%) as a colorless oil. [a]
D
+47.5 (c 1.5, CHCl3). 1H NMR: (CDCl3) d 8.01–7.02 (m, 25H, PhH),
5.49 (dd, 1H, J = 6.1 Hz, J = 2.9 Hz, H-2), 5.30 (br d, 1H, J = 6 Hz, H-
1), 4.91 (d, 1H, J = 3.5 Hz, H-1), 4.80 (d, 1H, J = 11.3 Hz, PhCH2),
4.78 (d, 1H, J = 12.2 Hz, PhCH2), 4.72–4.66 (m, 3H), 4.47–4.39 (m,

R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2359
2H), 4.18 (dd, 1H, J = 6.0 Hz, J = 2.9 Hz), 4.06–3.80 (m, 10H), 3.37 (t,
1H, J = 3.4 Hz), 2.91 (dd, 1H, J = 8.5 Hz, J = 5.1 Hz), 2.73–2.56 (m,
2H, SCH2), 1.62–1.57 (m, 2H, CH2), 1.37–1.04 (m, 40H, CH, CH2,
CH3), 0.87 (t, 3H, J = 6.4 Hz, CH3). 13C NMR (CDCl3): d 165.5
(C@O), 139.0, 138.7, 138.2, 133.0, 130.4, 129.8, 128.5, 129.4 (2C),
128.3, 128.2, 128.1, 127.8, 127.7, 127.6, 127.5, 127.3 (PhC), 99.4
(C-1), 79.1, 78.7, 76.7, 75.9, 75.1, 74.8 (PhCH2), 74.1 (PhCH2), 73,
1 (PhCH2), 72.9 (PhCH2), 70.1, 68.0, 67.8 (C-6), 61.4 (C-6), 31.8,
29.9,.29.1, 28.9, 22.6 (CH2), 17.4, 17.3, 17.1, 16.8, 16.7, 14.0 , 13.1,
12.9, 12.8, 12.7 (CH, CH3). Anal. Calcd for C67H93O12SSi2
(1197.70): C, 67.19; H, 7.83; S, 2.68. Found: C, 67.10; H, 7.61; S,
2.80.
(C-6g), 67.1 (C-4), 66.7 (C-40), 66.5 (C-6), 62.7 (C-60), 31.7, 31.3,
29.3, 29.2 (2C), 28.8, 22.6 (CH2), 14.0 (CH3). Anal. Calcd for
C89H92O20S (1513.74): C, 70.62; H, 6.13. Found: C, 70.54; H, 6.15.
3.28. 2,3,4,6-Tetra-O-benzoyl-
[2,3,4,6-tetra-O-benzyl- -galactopyranosyl-(1?3)]-2-O-
benzoyl-4-O-(1-fluoro-1,1,3,3-tetraisopropyl-1,3-disiloxane-3-
yl)-1-thio- -mannopyranosyl trichloroacetimidate (31)
a-D-mannopyranosyl-(1?6)-
a
-D
a-D
Treatment of compound 29 (355 mg, 0.2 mmol) with NBS
(53 mg, 0.3 mmol) in aqueous acetone (1% water content, 10 mL)
as described for the preparation of compound 11 and chromatogra-
phy with 15:1 toluene–ethyl acetate gave a colorless intermediate
which was treated with trichloroacetonitrile (0.5 ml) and K2CO3
(0.5 g) in dichloromethane (10 mL) as described for the preparation
of compound 12 and chromatography with 30:1 toluene–ethyl
acetate gave 31 (334 mg, 95%) as a colorless oil which was used
3.26. Octyl 2,3,4,6-tetra-O-benzoyl-
[2,3,4,6-tetra-O-benzyl- -galactopyranosyl-(1?3)]-2-O-ben-
zoyl-4-O-(1-fluoro-1,1,3,3-tetraisopropyl-1,3-disiloxane-3-yl)-
1-thio- -mannopyranoside (29)
a-D-mannopyranosyl-(1?6)-
a-D
a-D
for the next step without further characterization. [
a
]
D
+8.8 (c
Treatment of 28 (681 mg, 0.57 mmol), AgOTf (161 mg,
0.27 mmol), 4 Å molecular sieves (1 g) in dichloromethane
0.4, CHCl3). Anal. Calcd for C95H103Cl3FNO22Si2 (1792.36): C,
63.66; H, 5.79; N, 0.78. Found: C, 63.55; H, 5.74; N, 0.80.
(10 mL) with 15 (435 mg, 0.66 mmol) and sym-collidine (61 lL,
0.46 mmol) in dichloromethane (5 mL) at ꢀ25 °C as described for
the preparation of 16 and chromatography with 5:1 n-hexane–
ethyl acetate gave 29 (741 mg, 73%) as a colorless glassy material.
3.29. 2,3,4,6-Tetra-O-benzyl-
anhydro-2-O-benzoyl-b- -mannopyranoside (33) and octyl
2,3,4,6-tetra-O-benzoyl-1-thio- -mannopyranoside (34)
a-D-galactopyranosyl-(1?3)-1,6-
D
a-D
[a]
+25.3 (c 1.3, CHCl3). 1H NMR: (CDCl3): not assignable due to
D
extensive overlapping of signals. 13C NMR (CDCl3): d 166.1, 165.7,
165.3, 164.9 (C@O), 138.8, 137.9, 133.2, 133.1, 132.9, 132.8,
130.1, 130.0, 129.8, 129.7, 129.6, 129.3, 129.2, 128.5, 128.4, 128.3
(2C), 128.2, 128.1, 128.0, 127.8, 127.6, 127.4, 127.3, 127.1 (PhC),
79.2, 77.2, 76.0, 74.9, 74.8 (PhCH2), 73.1 (PhCH2), 72.9 (PhCH2),
70.3, 70.2, 68.5, 67.6 (C-6), 66.7, 62.7 (C-6), 31.7,28.9, 22.6 (CH2),
17.9, 17.7, 17.2, 17.1, 17.0, 16.8, 16.7, 14.0, 12.7, 12.5 (CH, CH3).
Anal. Calcd for C101H119FO21SSi2 (1776.25): C, 68.29; H, 6.75; S,
1.81. Found: C, 68.15, H, 6.75, S, 1.9.
BF3 etherate (0.2 mL, 1.6 mmol) was added at rt to a solution of
21 (1.2 g, 1.02 mmol) and 3212 (0.61 g, 1.02 mmol) and the mixture
was stirred for 1.5 h, poured into water, and extracted with dichlo-
romethane (3 ꢁ 50 mL). The combined extracts were washed with
aqueous NaHCO3 solution, dried and concentrated. Treatment of
the residue with Bu4NF trihydrate (50 mg) in THF (10 mL) as de-
scribed for the preparation of compound 17 and chromatography
with a 70:1–5:1 gradient toluene–ethyl acetate gave first 34
(476 mg, 64%). [
a
]
D
ꢀ2.5 (c 0.8, CHCl3). 1H NMR (CDCl3) d 8.11–
7.19 (m, 20H, PhH), 6.14 (t, 1H, J3,4 = J4,5 = 9.8 Hz, H-4), 5.85 (dd,
1H, J2,3 = 3.2 Hz, H-3), 5.81 (dd, 1H, J1,2 = 1.5 Hz, H-2), 5.55 (d, 1H,
H-1), 4.84 (ddd, 1H, J5,6a = 2.5 Hz, J5,6b = 4.5 Hz, H-5), 4.68 (dd, 1H,
J6a,6b = ꢀ12.2 Hz, H-6a), 4.53 (dd, 1H, H-6b), 2.80–2.62 (m, 2H,
SCH2), 1.72–1.61 (m, 2H, CH2), 1.38–1.12 (m, 10H, CH2), 0.88 (t,
3H, J = 6.8 Hz, CH3). 13C NMR (CDCl3): d 166.1, 165.4 (2C), 165.3
(C@O), 133.4 (2C), 133.2, 133.0, 129.8, 129.7, 129.3, 128.9 (2C),
128.6, 128.4 (2C), 128.3 (PhC), 82.5 (C-1), 72.1 (C-2), 70.5 (C-3),
69.2 (C-5), 67.1 (C-4), 62.9 (C-6), 31.7, 31.4, 29.5, 29.1, 29.0, 28.8,
22.6 (CH2), 14.1 (CH3). Anal. Calcd for C42H44O9S (724.86): C,
69.59; H, 6.12. Found: C, 69.43; H, 6.12.
3.27. Octyl 2,3,4,6-tetra-O-benzoyl-
[2,3,4,6-tetra-O-benzyl- -galactopyranosyl-(1?3)]-2-O-ben-
zoyl-1-thio- -mannopyranoside (30)
a-D-mannopyranosyl-(1?6)-
a-D
a-D
Treatment of 29 (60 mg, 0.03 mmol) with Bu4NF trihydrate
(10 mg) in THF (5 mL) as described for the preparation of 17 and
chromatography with 30:1 toluene–ethyl acetate gave 30 (46 mg,
89%) as a colorless oil. [
a]
+22.9 (c 0.7, CHCl3). 1H NMR (CDCl3):
D
d 8.14–7.18 (m, 45H, PhH), 6.16 (t, 1H, J3 ,4 = 10.1 Hz, H-40), 5.96
0
0
(dd, 1H, J2 ,3 = 3.2 Hz, H-30), 5.80 (dd, 1H, J1 ,2 = 1.8 Hz, H-10), 5.59
(dd, 1H, J1,2 = 1.2 Hz, J2,3 = 3.2 Hz, H-2), 5.41 (s, 1H, H-1), 5.21 (d,
1H, H-10), 4.99 (d, 1H, J1g,2g = 3.8 Hz, H-1g), 4.87 (d, 1H, J = –
11.6 Hz, PhCH2), 4.83 (d, 1H, J = ꢀ11.5 Hz, PhCH2), 4.72–4.66 (m,
4H, PhCH2, H-6a0), 4.55–4.45 (m, 2H, JPhCH2 = ꢀ10.3 Hz, H-
0
0
0
0
Eluted next was 33 (408 mg, 51%). [a]
+15.3 (c 0.6, CHCl3). 1H
D
NMR (CDCl3): d8.08–7.04 (m, 25H, PhH), 5.50 (br s, 1H, H-1), 5.09
(dd, 1H, J1,2 = 2.0 Hz, J2,3 = 5.3 Hz, H-2), 4.88 (d, 1H, J = ꢀ11.9 Hz,
PhCH2), 4.88 (d, 1H, J1gal,2gal = 3.4 Hz, H-1gal), 4.84 (d, 1H,
J = ꢀ11.5 Hz, PhCH2), 4.75 (br s, 2H, PhCH2), 4.61 (d, 1H,
J = ꢀ11.3 Hz, PhCH2), 4.51 (d, J = 7.4 Hz, H-6a), 4.45 (d, 1H,
J = ꢀ11.2 Hz, PhCH2), 4.35 (br d, 1H, J = 5.4 Hz, H-5), 4.20–4.18
(m, 1H, H-3), 4.06–3.92 (m, 6H, H-20, H-30, H-40, H-50, PhCH2),
3.83 (br d, 1H, J = 8.5 Hz, H-4), 3.59 (dd, 1H, J6a,6b = 6.9 Hz,
50,PhCH2), 4.45 (dd, 1H, J6a ,6b = ꢀ11.8 Hz, J5,6b = 7.4 Hz, H-6b0),
4.40–4.29 (m, 3H, JPhCH2 = ꢀ11.8 Hz, JPhCH2 = ꢀ11.7 Hz, PhCH2, H-
5), 4.22 (t, 1H, J3,4 = J4,5 = 9.8 Hz, H-4), 4.16 (dd, 1H, J5,6a = 5.6 Hz,
J6a,6b = ꢀ11.3 Hz, H-6a), 4.06 (dd, 1H, J2g,3g = 10.4 Hz, H-2g), 3.96–
3.89 (m, 5H, H-3, H-3g, H-4g, H-5g, H-6b), 3.57 (t, 1H, J = 8.9 Hz,
H-6ag), 3.44 (dd, 1H, J6ag,6bg = ꢀ8.8 Hz, J5g,6bg = 5.3 Hz, H-6bg),
2.80–2.64 (m, 2H, SCH2), 1.73–1.63 (m, 2H, CH2), 1.25–1.20 (m,
10H, CH2), 0.83–0.78 (m, 3H, CH3). 13C NMR (CDCl3): d 166.2,
165.7, 165.4, 165.3, 165.1 (C@O), 138.7, 138.4, 138.2, 137.7,
133.3, 133.2, 133.1, 132.9, 129.9, 129.8, 129.7 (2C), 129.4, 129.2,
129.0, 128.6, 128.5 (3C), 128.4 (2C), 128.3 (2C), 128.2, 128.1,
128.0, 127.9, 127.7, 127.5, 127.4, 127.3 (PhC), 101.9 (C-1g), 97.6
(C-10), 82.6 (C-1), 81.9 (C-3), 79.5 (C-3g), 76.3 (C-2g), 74.7 (PhCH2),
74.4 (PhCH2), 74.2 (C-5g), 73.5 (C-2), 73.2 (PhCH2), 72.4 (PhCH2),
71.7 (C-5), 70.3 (C-20), 70.2 (C-30), 69.7 (C-4g), 68.7 (C-50), 67.9
0
0
0
J6b,5 = 6.3 Hz, H-6), 3.32 (t, J5 ,6a = J6a ,6b = 8.6 Hz, H-6a0), 2.88 (dd,
0
0
0
0
1H, J5 ,6b = 5.4 Hz, H-6b0), 2.61 (d, 1H, J = 8.7 Hz, OH). 13C NMR
(CDCl3): d 165.7 (C@O), 138.7, 138.6, 138.3, 137.9, 133.2, 129.8,
129.4, 129.0, 128.4 (2C), 128.3, 128.2, 128.1 (2C), 127.9 (2C),
127.8, 127.5 (2C), 127.4 (2C), 125.3 (PhC), 99.6 (C-1), 99.0 (C-
0
0
1gal), 78.8 (C-2gal), 76.9 (C-3), 76.5 (C-5), 76.4 (C-3gal), 74.8
(PhCH2), 74.6 (C-4gal), 74.3 (PhCH2), 72.9 (PhCH2), 72.3 (PhCH2),
70.1 (C-4), 69.6 (C-5gal), 69.4 (C-2), 67.6 (C-6gal), 65.1 (C-6). Anal.
Calcd for C47H48O11 (788.88): C, 71.56; H, 6.13. Found: C, 71.20;
H, 6.16.

2360
R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
3.30. Octyl 2,3,4,6-tetra-O-benzoyl-
a
-
D
-mannopyranosyl-(1?2)-
(2H, H-4, H-4Gal), 3.84 (2H, H-6a00, H-6a0), 3.81 (4H, H-5, H-6aGlc
,
H-6bGlc, H-3Gal), 3.78 (2H, H-3, H-2Gal), 3.77 (H-6aGal), 3.76 (H-
6b), 3.71 (2H, H-300, H-500), 3.68 (3H, H-6b0, H-6b00, H-6bGal), 3.66
(H-3Glc), 3.63 (H-50), 3.62 (H-40), 3.58 (2H, H-400, H-4Glc), 3.23 (dd,
1H, J2Glc,3Glc = 10.4 Hz, H-2Glc), 2.68 (t, 2H, J = 7.3 Hz, SCH2), 1.68–
1.62 (m, 2H, CH2), 1.45–1.36 (m, 2H, CH2), 1.30–1.29 (m, 8H,
CH2), 0.89 (t, 3H, J = 6.9 Hz, CH3). 13C NMR (MeOD-d4): d 104.1
(C-100), 103.2 (C-1), 102.4 (C-1Gal), 100.1 (C-10), 85.9 (C-1Glc), 81.5
(C-3), 80.4 (C-20), 78.6 (C-4Glc), 74.9 (C-500), 74.5 (C-50), 74.3 (C-5),
74.0 (C-3Glc), 73.5 (C-5Glc), 73.0 (C-5Gal), 72.4 (C-300), 72.1 (C-30),
71.9 (C-200), 71.5 (C-3Gal), 71.3 (C-4Gal), 71.2 (C-2), 70.8 (C-2Gal),
69.0 (C-40), 68.8 (C-400), 67.8 (C-6), 67.2 (C-4), 63.1 (C-6Gal), 63.0
(C-60), 62.9 (C-600), 62.4 (C-6Glc), 56.4 (C-2Glc), 33.0, 32.6, 30.9,
30.4, 30.3, 29.9, 23.7 (CH2), 14.5 (CH3). HRMS ESI Calcd for
3,4,6-tri-O-acetyl-
a
-
D
-mannopyranosyl-(1?6)-[2,3,4,6-tetra-O-
-manno-
benzyl- -galactopyranosyl-(1?3)]-2-O-benzoyl-a-D
a-
D
pyranosyl-(1?4)-2-azido-3,6-di-O-benzyl-2-deoxy-1-thio-a-D-
glucopyranosid (36)
TMSOTf (5
l
L) was added at ꢀ20 °C to a solution of 25 (270 mg,
0.17 mmol) and 3512 (196 mg, 0.19 mmol) in diethylethylether
(10 mL). After 45 min an additional amount of 35 (50 mg,
0.05 mmol) was added and stirring at ꢀ20 °C continued for 1.5 h.
Workup as described for the preparation of compound 13 followed
by treatment of the intermediate (252 mg, 60%) with Bu4NF trihy-
drate (50 mg) in THF (10 mL) as described for the preparation of 17
and chromatography with 5:1 toluene-ethyl acetate gave 36
(192 mg, 51%). [
a]
+23.5 (c 0.4, CHCl3). 1H NMR (CDCl3): d 8.09–
C
38H70NO24S (M+H+): m/z 956.393; Found: m/z 956.397.
D
7.04 (m, 55H, PhH), 6.11 (t, 1H, J3 ,4 = J4 ,5 = 10.0 Hz, H-400), 5.94
00 00
00 00
(dd, 1H, J2 ,3 = 3.2 Hz, H-300), 5.68 (dd, 1H, J1 ,2 = 1.9 Hz, H-200),
5.54 (dd, 1H, J1,2 = 1.5 Hz, J2,3 = 2.9 Hz, H-2), 5.47 (d, 1H, H-1),
Acknowledgments
00 00
00 00
5.46 (t, 1H, J3 ,4 = J4 ,5 = 9.6 Hz, H-40), 5.40 (d, 1H, J1glc,2glc = 5.4 Hz,
We thank the following colleagues for their support of this
work: K. Albert and his crew for recording the NMR spectra; K.-P.
Zeller and his crew for performing the mass-spectroscopy; P. Krue-
ger for performing the elemental analyses. This work was finan-
cially supported by the Deutsche Forschungsgemeinschaft and
the Fonds der Chemischen Industrie.
0
0
0
0
H-1glc), 5.39 (dd, J2 ,3 = 2.9 Hz, H-30), 5.03 (d, 1H, J1 ,2 = 1.2 Hz, H-
10), 5.01 (d, 1H, H-100), 4.97 (d, 1H, J1gal, 2gal = 3.8 Hz, H-1gal), 4.87
(d, 1H, J = 11.6 Hz, PhCH2), 4.82 (d, 1H, J = 10.7 Hz, PhCH2), 4.80
(d, 1H, J = 11.5 Hz, PhCH2), 4.76 (d, 1H, J = 11.3 Hz, PhCH2), 4.72
(d, 1H, J = 11.6 Hz, PhCH2), 4.62 (s, 2H, PhCH2), 4.56 (H-6a00), 4.55
(2H, PhCH2), 4.52 (H-500), 4.48 (d, 1H, J = 11.3 Hz, PhCH2), 4.40 (H-
6b00), 4.31 (d, 1H, J = 11.8 Hz, PhCH2),4.27 (d, 1H, J = 11.8 Hz,
PhCH2), 4.25 (H-5glc), 4.25 (H-5glc), 4.21 (H-6a0), 4.08 (H-6b0), 4.06
(H-50), 4.04 (H-2Gal), 4.03 (H-20), 4.02 (H-4), 3.92 (H-4Glc), 3.90 (H-
0
0
0
0
References
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Gal), 3.89 (H-5, H-2Glc), 3.88 (H-4Gal), 3.87 (H-3Gal), 3.85 (H-3), 3.84
(H-6aGlc), 3.81 (H-6a), 3.77 (H-6bGlc), 3.74 (H-3Glc), 3.61 (br d, 1H,
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1.25 (m, 12H, CH2), 0.86 (br t, 3H, J = 7.0 Hz, CH3). 13C NMR (CDCl3):
d 171.0, 170.3, 169.5, 165.7, 165.6, 165.2 (2C), 165.0 (C@O), 138.7,
138.4, 138.2 (2C), 137.7, 137.5, 133.5, 133.4, 133.2, 133.1, 132.9,
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(2C), 127.7, 127.5 (3C), 127.4 (2C), 127.3 (PhC), 102.1 (C-1Gal),
99.3 (C-100), 98.7 (C-1), 98.3 (C-10), 83.3 (C-1Glc), 81.5 (C-3Glc),
81.3 (C-3), 79.5 (C-3Gal), 77.4 (C-20), 76.3 (C-2Gal), 75.0 (C-4Glc),
74.9 (PhCH2), 74.2 (C-4Gal), 73.4 (PhCH2), 73.1 (PhCH2), 72.3 (2C,
C-5, PhCH2), 71.9 (C-2), 70.8 (C-5Glc), 70.7 (C-200), 70.4 (C-30), 69.6
(C-5Gal), 69.4 (2C, C-300, C-500), 69.3 (C-6Glc), 68.4 (C-50), 67.8 (C-
12. Ziegler, T.; Dettmann, R.; Duszenko, M.; Kolb, V. Carbohydr. Res. 1996, 295, 7–
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Gal), 67.0 (C-400), 66.6 (C-4), 66.4 (C-6), 66.2 (C-40), 64.2 (C-2Glc),
62.8 (C-600), 62.1 (C-60), 31.8, 30.5, 29.6, 29.2 (2C), 29.0, 22.7
(CH2), 14.1 (CH3). Anal. Calcd for C121H129N3O32S (2169.39): C,
66.99; H, 5.99; N, 1.94. Found: C, 66.89; H, 6.23; N, 1.81.
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(1?4)-2-amino-2-deoxy-1-thio-
a
-
D
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ˇ
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J2 ,3 = 3.1 Hz, H-200), 3.93 (H-20), 3.92 (H-6a), 3.90 (H-30), 3.87
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R. Dettmann, T. Ziegler / Carbohydrate Research 346 (2011) 2348–2361
2361
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