Convergent syntheses of LeX analogues

Article abstract of DOI:10.3762/bjoc.6.17

The synthesis of three Le^x derivatives from one common protected trisaccharide is reported. These analogues will be used respectively for competitive binding experiments, conjugation to carrier proteins and immobilization on gold. An N-acetylglucosamine monosaccharide acceptor was first glycosylated at O-4 with a galactosyl imidate. This coupling was performed at 40 °C under excess of BF₃·OEt₂ activation and proceeded best if the acceptor carried a 6-chlorohexyl rather than a 6-azidohexyl aglycon. The 6-chlorohexyl disaccharide was then converted to an acceptor and submitted to fucosylation yielding the corresponding protected 6-chlorohexyl Lex trisaccharide. This protected trisaccharide was used as a precursor to the 6-azidohexyl, 6-acetylthiohexyl and 6-benzylthiohexyl trisaccharide analogues which were obtained in excellent yields (70-95%). In turn, we describe the deprotection of these intermediates in one single step using dissolving metal conditions. Under these conditions, the 6-chlorohexyl and 6-azidohexyl intermediates led respectively to the n-hexyl and 6-aminohexyl trisaccharide targets. Unexpectedly, the 6-acetylthiohexyl analogue underwent desulfurization and gave the n-hexyl glycoside product, whereas the 6-benzylthiohexyl analogue gave the desired disulfide trisaccharide dimer. This study constitutes a particularly efficient and convergent preparation of these three Le^x analogues.

Full text of DOI:10.3762/bjoc.6.17

Convergent syntheses of LeX analogues
An Wang, Jenifer Hendel and France-Isabelle Auzanneau*
Full Research Paper
Open Access
Address:
Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
Department of Chemistry, University of Guelph, Guelph, Ontario, N1G
2W1, Canada
doi:10.3762/bjoc.6.17
Received: 10 August 2009
Accepted: 21 January 2010
Published: 22 February 2010
Email:
France-Isabelle Auzanneau* - fauzanne@uoguelph.ca
* Corresponding author
Guest Editor: T. K. Lindhorst
Keywords:
© 2010 Wang et al; licensee Beilstein-Institut.
License and terms: see end of document.
Birch reduction; convergent synthesis; desulfurization; Lewis X
Abstract
The synthesis of three Lex derivatives from one common protected trisaccharide is reported. These analogues will be used respect-
ively for competitive binding experiments, conjugation to carrier proteins and immobilization on gold. An N-acetylglucosamine
monosaccharide acceptor was first glycosylated at O-4 with a galactosyl imidate. This coupling was performed at 40 °C under
excess of BF3·OEt2 activation and proceeded best if the acceptor carried a 6-chlorohexyl rather than a 6-azidohexyl aglycon. The
6-chlorohexyl disaccharide was then converted to an acceptor and submitted to fucosylation yielding the corresponding protected
6-chlorohexyl Lex trisaccharide. This protected trisaccharide was used as a precursor to the 6-azidohexyl, 6-acetylthiohexyl and
6-benzylthiohexyl trisaccharide analogues which were obtained in excellent yields (70–95%). In turn, we describe the deprotection
of these intermediates in one single step using dissolving metal conditions. Under these conditions, the 6-chlorohexyl and 6-azido-
hexyl intermediates led respectively to the n-hexyl and 6-aminohexyl trisaccharide targets. Unexpectedly, the 6-acetylthiohexyl
analogue underwent desulfurization and gave the n-hexyl glycoside product, whereas the 6-benzylthiohexyl analogue gave the
desired disulfide trisaccharide dimer. This study constitutes a particularly efficient and convergent preparation of these three Lex
analogues.
Introduction
Our group is involved in the design of new anti-cancer vaccines ture that deal with the chemical [12-36] or chemoenzymatic
based on the Tumor Associated Carbohydrate Antigen (TACA) [37,38] preparation of Lex analogues as well as that of Lex
dimeric Lex (dimLex) [1-6]. This tumor specific antigen intermediate building blocks to be further converted into the
consists of a hexasaccharide that displays the Lex trisaccharide Sialyl Lex tetrasaccharide. The chemical syntheses usually
antigen linked to O-3″ of the galactose residue of another Lex follow one of three synthetic schemes: 1. a stepwise approach
trisaccharide. Since it was first characterized [7,8], the Lex anti- involving the successive galactosylation then fucosylation of a
genic determinant, β-D-Galp(1,4)[α-LFucp(1,3)]-D-GlcNAcp, glucosamine acceptor [12-28]; 2. a stepwise approach in which
has been found on numerous cells and tissues such as kidney the sequence of glycosylation of the glucosamine acceptor is
tubules, gastrointestinal epithelial cells, and cells of the spleen reversed, i.e. the fucosylation is followed by the galactosylation
and brain [9-11]. Thus, there are numerous reports in the litera- [28-34]; 3. a block approach in which a lactosamine derivative
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
Figure 1: Structure of Lex analogues 1–3.
Figure 2: Monosaccharide glycosyl acceptors (4–6) and donors (7–9)
used in this study.
prepared from lactose is subjected to fucosylation at O-3
[35,36]. Whereas these reports usually describe the preparation
of one compound to be used in a specific experiment, we
describe here the convergent synthesis of the three Lex deriva- glycosylation at O-4 of glucosamine glycosyl acceptors with
tives 1–3 (Figure 1) from one common protected trisaccharide galactosyl donor 8, which is chloroacetylated rather than acet-
intermediate. These three Lex analogues (1–3) will be used ylated at O-3. Finally, we also investigated the reactivity
respectively for competitive binding experiments (1), conjuga- towards glycosylation of the N-acetylated and phthalimido
tion to carrier proteins (2) and immobilization to a gold plate acceptors 5 and 6, respectively, that both carry a 6-azidohexyl
(3).
aglycon (Figure 2).
Results and Discussion
Synthesis of monosaccharide building blocks. The 6-chloro-
Our synthetic approach to prepare these Lex derivatives began hexyl acceptor 4 was prepared in four steps from the known
with the galactosylation at O-4 of glycosyl acceptor 4 with the [43] chlorohexyl glucoside 10 (Scheme 1). Thus, peracetate 10
known [39-41] galactosyl donor 7 followed by deprotection at was deacetylated (NaOMe/MeOH) and converted to the
O-3 of the glucosamine residue and fucosylation of the resulting benzylidene acetal 11 by reaction with benzaldehyde dimethyl
disaccharide with the known [42] ethylthioglycoside 9. Since in acetal under camphorsulfonic acid (CSA) catalysis. Chloro-
addition to the Lex trisaccharide we are also interested in acetylation of alcohol 11 gave the intermediate 12 which was
preparing fragments of the dimLex antigen, we examined the converted to acceptor 4 via the reductive opening of the
Scheme 1: Synthesis of monosaccharide glycosyl acceptors 4–6.
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
benzylidene acetal using NaCNBH3 and HCl·Et2O in anhyd- AcOH at 70 °C to remove the isopropylidene group affording
rous THF at 0 °C.
diol 22. The diol 22 was selectively acetylated at O-4 by
converting it to the corresponding cyclic methylorthoacetate
Both 6-azidohexyl acceptors 6 and 5 were prepared from the and opening the orthoacetate in situ by adding water to the reac-
anomeric mixture of the known tetraacetate 13 [44]. Thus, tion mixture. The resulting triacetate was chloroacetylated at
tetraacetate 13 was reacted with 6-chlorohexanol (4 equiv) in O-3 and the resulting fully protected thioglycoside 23 was
the presence of BF3·OEt2 (5 equiv). To promote coupling, the converted to the corresponding hemiacetal that was, in turn,
reaction mixture was either stirred for 1 h at 50 °C in an oil bath treated with trichloroacetonitrile and DBU to give the
(Supporting Information File 1, Method A) or submitted to α-trichloroacetimidate galactosyl donor 8.
microwave irradiation for 5 min at 50 °C (Supporting Informa-
tion File 1, Method B). After acetylation of the excess chloro- Glycosylation at O-4 of glucosamine acceptors. It is well
hexanol to ease its removal, pure glycoside 14 was isolated in known that the hydroxyl group at C-4 of N-acetylglucosamine
excellent yield whether method A or B was followed. Thus, is a poor nucleophile and has reduced reactivity towards glyco-
these syntheses of glycoside 14 constitute efficient alternatives sylation when compared to other acceptors [48-50]. However,
to that reported by Nitz et al. in which the starting material was we have recently reported the successful O-4 glucosylation of
the corresponding anomeric bromide [45]. Nucleophilic an N-acetylglucosamine monosaccharide acceptor using a
displacement of the chlorine atom in glycoside 14 (NaN3, DMF, peracetylated glucopyranose α-trichloroacetimidate donor under
80 °C) gave the known [46] 6-azidohexyl glycoside 15 quantita- activation with 2 equiv of BF3·OEt2 at room temperature [51].
tively. Zemplén deacetylation of triacetate 15 followed by We applied similar conditions: 2 equiv BF3·OEt2, 5 equiv of
conversion of the triol to the 4,6-benzylidene acetal (16) and donor, 1 h at 40 °C for the coupling of donors 7 and 8 with the
then chloroacetylation at O-3 gave intermediate 17 that was acceptors 4-6 (Table 1). As can be seen in Table 1 the 6-chloro-
submitted to reductive opening of the benzylidene group hexyl glycoside acceptor 4 was easily glycosylated with either
(NaCNBH3, HCl·Et2O) to yield acceptor 6.
donors 7 or 8, affording the desired disaccharides 24 and 25 in
about 70% yield for both reactions (entries 1 and 2).
The triacetate 15 was also converted in seven steps to acceptor
5. The phthalimido group was first removed (ethylenediamine, In contrast, the coupling of donor 8 with the 6-azidohexyl
EtOH) and the free amine acetylated. Zemplén deacetylation glycoside acceptor 5 did not proceed well (entry 3). Monitoring
was followed by conversion of the triol to the 4,6-benzylidene of the reaction by TLC showed degradation of the acceptor, and
acetal 18 which was chloroacetylated at O-3 to give the fully isolation of the desired disaccharide required both silica gel
protected intermediate 19. Finally, the benzylidene acetal in
compound 19 was reductively opened with Et3SiH and TfOH in
Table 1: Glycosylation at O-4 of glucosamine acceptors 4–6a.
CH2Cl2 at −30 °C to give acceptor 5.
The trichloroacetimidate glycosyl donor 8 was prepared from
the p-thiotolyl glycoside 20 [47] (Scheme 2). Diol 20 was first
acetylated to the diacetate 21 which was then treated with 90%
Entry
Donor
Acceptor
Product (%)
1
2
3
4
7
8
8
8
4
4
5
6
24 (69%)
25 (72%)
26 (27%)b
27 (11%)
aReagents and conditions: BF3·OEt2 (2 equiv), donor (5 equiv), CH2Cl2,
40 °C, 1 h.
bContaminated with degraded acceptor.
Scheme 2: Synthesis of the galactosyl donor 8.
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
chromatography and RP-HPLC. Indeed, despite our efforts, and
even though its structure was confirmed by NMR and HR-ESI
mass spectrometry, disaccharide 26 could not be isolated free of
degraded acceptor and/or disaccharide. To further test if the
N-acetyl group was impacting negatively the glycosylation of
acceptor 5, we attempted to couple trichloroacetimidate 8 with
the phthalimido acceptor 6. However as can be seen in Table 1,
entry 4, this glycosylation also gave disappointing results: TLC
showed a considerable amount of degraded products and the
isolation of the desired disaccharide from the reaction mixture
required both silica gel chromatography and RP-HPLC. In this
case, the disaccharide 27 could be obtained pure albeit in very
low yield. These last two reactions suggest that the presence of
the azido group on the hexyl aglycon carried by acceptors 5 and
6 is not compatible with the glycosylation conditions that we
have established previously [51] for the glycosylation at O-4 of
glucosamine acceptors. The disaccharide 24 was further used in
the preparation of the Lex analogues 1–3.
Preparation of protected Lex analogues. The chloroacetate in
disaccharide 24 was removed with thiourea (C5H5N/EtOH, 70
°C) to give the acceptor disaccharide 28 (61%), which was then
Scheme 3: Convergent synthesis of trisaccharides 29–32.
fucosylated with the thioethyl glycoside 9 under copper (II) bro- chloride 29 was allowed to react for 16 h with excess benzyl-
mide–tetrabutylammonium bromide activation (Scheme 3). The thiol (15 equiv) and sodium hydride (15 equiv) in DMF at 80
desired Lex trisaccharide 29 was obtained in excellent yield and °C. These reaction conditions led to the displacement of the
the α-configuration of the newly formed fucosidic bond was chloride as well as to some deacetylation of the galactose
confirmed by 1H NMR (JH-1′,H-2′ = 3.7 Hz). The 6-chlorohexyl residue. Thus, after acetylation of the crude product, the desired
trisaccharide glycoside 29 was in turn used as a precursor to the 6-benzylthiohexyl trisaccharide 32 was isolated in excellent
6-azidohexyl, 6-acetylthiohexyl and 6-thiobenzylhexyl trisac- yield (Scheme 3). It is important to point out that the 6-chloro-
charides 30–32 (Scheme 3).
hexyl glycoside 29 and the 6-benzylthiohexyl glycoside 32
co-eluted on silica gel and that only a very careful analysis of
Thus, nucleophilic displacement of the chloride with sodium the NMR data recorded for the product could confirm the
azide or potassium thioacetate was carried out in DMF at 80 °C absence of unreacted starting material. Indeed, the large excess
and provided the 6-azidohexyl and 6-acetylthiohexyl trisacchar- of benzylthiolate used to displace the chloride in trisaccharide
ides 30 and 31, respectively. The introduction of the azido or 29 was essential for its complete conversion to the desired
thioacetyl groups into trisaccharides 30 and 31 was confirmed 6-benzylthiohexyl glycoside 32. The structure of trisaccharide
by HR-ESI mass spectrometry and by NMR. Indeed, the signals 32 was confirmed by HR-ESI MS as well as by NMR. The
assigned to the methylene CH2Cl in trisaccharide 29 (1H NMR methylene CH2SBn gave signals at 2.36 and 31.3 ppm, in the
δ 3.50 ppm, 13C NMR δ 44.9 ppm) were no longer observed in 1H and 13C NMR spectra, respectively whereas the S-benzyl
trisaccharides 30 and 31. The methylene CH2N3 in tri- group gave additional signals in the aromatic regions as well as
saccharide 30 gave signals at 3.20 and 54.3 ppm in the 1H and signals corresponding to the SCH2Ph methylene around 3.70
13C NMR spectra, respectively, whereas the methylene and 36.3 ppm in the 1H and 13C NMR spectra, respectively.
CH2SAc in trisaccharide 31 gave signals at 2.81 and around
28.5 ppm, in the 1H and 13C NMR spectra, respectively. In add- Deprotection of trisaccharides 29–32 under dissolving metal
ition, signals corresponding to the thioacetyl group in tri- conditions. As reported by Seeberger et al. [52], the removal of
saccharide 31 were identified at 2.29 ppm and 30.6 ppm in the O- and S-benzyl groups as well as that of O-acetyl groups can
1H and 13C NMR spectra, respectively. Since, as will be be accomplished in one step and concurrently with the reduc-
described below, the deprotection of trisaccharide 31 under tion of azido groups to the corresponding amines, using Birch
dissolving metal conditions did not provide the desired tri- reduction conditions. Thus we embarked on the one step depro-
saccharide 3, the 6-benzylthiohexyl glycoside 32 was also tection of trisaccharides 29–32 with sodium in ammonia
prepared from the 6-chlorohexyl glycoside 29. Thus, the (Table 2).
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
Table 2: One step deprotection of trisaccharides 29–32a.
Scheme 4: Proposed mechanism for the desulfurization of thioacetate
31 under dissolving metal conditions.
Entry
Trisaccharide Product
Yield (%)
the formation of intramolecular Lex–Lex interactions such as
those reported by de la Fuente and Penadés for a similar
analogue [33]. Following published procedures, the disulfide
dimer 3 will be reduced immediately prior to its conjugation to
proteins [53] or immobilization on gold surface or gold nano-
particules [34].
1
2
3
4
29
30
31
32
1
2
1
3
82
59
73
70
aReagents and conditions: Na/NH3(l), −78 °C, 50 min.
In conclusion, we have reported above the efficient and conver-
Treatment of trisaccharides 29 and 30 with sodium in liquid gent preparation of three Lex derivatives (1–3) from one
ammonia at −78 °C followed by neutralization of the reaction common protected trisaccharide (29). Our results seem to indi-
mixtures with AcOH gave the desired trisaccharides 1 and 2 cate that glycosylation at O-4 of a glucosamine monosac-
(entries 1 and 2) that were isolated pure after chromatography charide acceptor under excess BF3·OEt2 activation at 40 °C is
on a Biogel P2 column eluted with water for compound 1, and compatible with a chlorinated aglycon but not with an aglycon
0.05 M ammonium acetate for the 6-aminohexyl compound 2. carrying an azido group. We have also established that the fully
Whereas the structure of trisaccharide 1 was confirmed by protected precursors could be deprotected in one single step to
HR-ESI mass spectrometry and NMR, the structure of the give the final target compounds using dissolving metal condi-
6-aminohexyl glycoside 2 was confirmed by comparing its tions. However, we observed that a thioacetylated derivative
analytical data to that previously reported [31]. To our surprise, will undergo an undesired reductive desulfurization. This study
treatment of the 6-acetylthiohexyl trisaccharide 31 under Birch constitutes a particularly efficient convergent preparation of
reduction conditions did not lead to the desired corresponding analogues that can each be used for a specific biochemical
thiol or disulfide product but produced the hexyl glycoside 1. application.
The mechanism proposed to explain this reductive desulfuriz-
ation is shown in Scheme 4. It involves first a single electron Experimental
transfer to the thioacetyl group that is followed by the cleavage General Methods: 1H (600.14, 400.13 or 300.13 MHz) and
of the carbon sulfur bond giving a thioacetate salt and an alkyl 13C NMR (150.9, 100.6 or 75.5 MHz) spectra were recorded for
radical. The alkyl radical is then converted to the correspond- compounds solubilized in CDCl3 (internal standard, for 1H:
ing anion by a second electron transfer and the resulting anion residual CHCl3 δ 7.24; for 13C: CDCl3 δ 77.0) or D2O [external
is protonated by ammonia giving trisaccharide 1.
standard 3-(trimethylsilyl)-propionic acid-d4, sodium salt (TSP)
for 1H δ 0.00, for 13C δ 0.00]. Chemical shifts and coupling
In contrast to the thioacetate 31, treatment of the 6-benzylthio- constants were obtained from a first-order analysis of one-
hexyl glycoside 32 under Birch reduction conditions did not dimensional spectra. Assignments of proton and carbon reson-
lead to desulfurization and gave the disulfide trisaccharide ances were based on COSY and 13C–1H heteronuclear correl-
dimer 3. Under these reductive conditions, and based on the ated experiments. Mass spectra were obtained under electron
work by Seeberger et al. [52], we did not expect the formation spray ionization (ESI) on a high resolution mass spectrometer.
of the disulfide dimer as the major product but rather that of the TLC were performed on precoated aluminum plates with Silica
corresponding thiol. However, the structure and homogeneity of Gel 60 F254 and detected with UV light and/or charred with a
disulfide dimer 3 was unequivocally confirmed by HR-ESI solution of 10% H2SO4 in EtOH. Compounds were purified by
mass spectrometry and NMR. Interestingly this dimer gave a flash chromatography with Silica Gel 60 (230–400 mesh) unless
well resolved 1H NMR spectrum in D2O that did not support otherwise stated. Solvents were distilled and dried according to
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
standard procedures [54], and organic solutions were dried over H-4, H-6b, H-3′, H-4″, OCHHCH2); 3.78 (d, 1H, J = 3.0 Hz,
Na2SO4 and concentrated under reduced pressure below 40 °C. H-4′); 3.73 (m, 2H, H-6a″, H-6b″); 3.70 (m, 1H, H-2′); 3.66 (m,
HPLC purifications were run with HPLC grade solvents.
1H, H-3″); 3.60 (m, 3H, H-5, H-5″, OCHHCH2); 3.59 (m, 1H,
H-2″); 2.99 (t, 2H, J = 7.0 Hz, CH2NH2); 2.03, 2.01 (s, 6H,
n-Hexyl 2-acetamido-2-deoxy-3-O-(α-L-fucopyranosyl)-4-O- CH3CO); 1.57, 1.67 (m, 4H, OCH2CH2, CH2CH2NH2);
(β-D-galactopyranosyl)-β-D-glucopyranoside (1). Tri- 1.30–1.42 (m, 4H, OCH2CH2CH2CH2); 1.17 (d, 3H, J = 6.0
saccharide 29 (20 mg, 0.017 mmol) or trisaccharide 31 (19 mg, Hz, H-6′). 13C-NMR (100 MHz, D2O): 173.96 (C=O); 101.64
0.016 mmol) were dissolved in THF (5 mL) and liquid (C-1″); 100.81 (C-1); 98.45 (C-1′); 75.16 (C-5); 74.73 (C-3,
ammonia (20 mL) was condensed into the solution at −78 °C. C-5″); 73.17 (C-4); 72.28 (C-3″); 71.70 (C-4′); 70.85 (C-2″);
Na (74 mg, 3.2 mmol) was added and the mixture was stirred 70.30 (OCH2CH2); 69.01 (C-3′); 68.15 (C-4″); 67.51 (C-2′);
for 50 min at −78 °C. The reaction was quenched with MeOH 66.53 (C-5′); 61.31 (C-6″); 59.57 (C-6); 55.65 (C-2); 39.21
(5 mL) and the ammonia was allowed to evaporate at room (CH2NH2); 28.18, 26.46, 25.05, 24.45 [OCH2(CH2)4]; 22.05
temp. The remaining solution was neutralized with acetic acid (CH3CO); 15.10 (C-6′). HRESIMS calcd for C26H48N2O15
(203 μL, 3.5 mmol), the solvent was evaporated and the residue [M+H]+ 629.3133, found 629.3121.
was passed twice through a Biogel P2 column (100 × 1 cm)
eluted with Milli-Q water to give the trisaccharide 1 (8.5 mg, 6,6′-Dithio-bis(hexan-1,6-diyl)-bis[2-acetamido-2-deoxy-3-
82% from 29; 7.0 mg, 73% from 31) as a white amorphous O-α-L-fucopyranosyl-4-O-(β-D-galactopyranosyl)-β-D-gluc-
powder after lyophilization. [α]D = −47 (c 0.5, MeOH), 1H opyranoside] (3). The 6-benzylthiohexyl trisaccharide 32 (30
NMR (400 MHz, D2O): δ 5.12 (d, 1H, J = 4.5 Hz, H-1′); 4.83 mg, 0.024 mmol) was deprotected in the same conditions as
(m, 1H, H-5′); 4.53 (d, 1H, J = 7.5 Hz, H-1); 4.46 (d, 1H, J = described above for the deprotection of trisaccharide 29. After
7.5 Hz, H-1″); 4.00 (dd, 1H, J = 12.0, 1.0 Hz, H-6a); 3.83–3.95 work up, the residue was passed through a Biogel P2 column
(m, 7H, H-2, H-3, H-4, H-6b, H-3′, H-4″, OCHHCH2); 3.78 (d, eluted with water to give the trisaccharide 3 (10.6 mg, 70%) as
1H, J = 3.0 Hz, H-4′); 3.73 (m, 2H, H-6a″, H-6b″); 3.70 (m, 1H, white amorphous powder after lyophilization. [α]D = −57 (c 0.7,
H-2′); 3.66 (m, 1H, H-3″); 3.60 (m, 3H, H-5, H-5″, MeOH), 1H NMR (600 MHz, D2O): δ 5.05 (d, 1H, J = 3.8 Hz,
OCHHCH2,); 3.59 (m, 1H, H-2″); 2.03 (s, 3H, CH3CO); 1.55 H-1′); 4.82–4.75 (m, 1H, H-5′); 4.47 (d, 1H, J = 7.7 Hz, H-1);
( m , 2 H , O C H 2 C H 2 ) ; 1 . 2 4 – 1 . 3 7 ( m , 6 H , 4.39 (d, 1H, J = 7.9 Hz, H-1″); 3.95 (d, 1H, J = 10.9 Hz, H-6a);
OCH2CH2CH2CH2CH2); 1.17 (d, 3H, J = 6.0 Hz, H-6′); 0.88 (t, 3.90–3.76 (m, 7H, H-2, H-3, H-4, H-6b, H-3′, H-4″,
3H, J = 6.6 Hz, CH2CH3). 13C-NMR (100 MHz, D2O): 174.17 OCHHCH2); 3.75–3.71 (m, 1H, H-4′); 3.70–3.56 (m, 4H,
(C=O); 101.81 (C-1″); 100.91 (C-1); 98.61 (C-1′); 75.32 (C-5); H-2′H-3″, H-6a″, H-6b″); 3.55–3.49 (m, 3H, H-5, H-5″,
74.94, 74.88 (C-3, C-5″); 73.36 (C-4); 72.44 (C-3″); 71.89 OCHHCH2); 3.44 (t, 1H, J = 8.1 Hz, H-2″); 2.70 (t, 2H, J = 7.1
(C-4′); 71.02 (C-2″); 70.66 (OCH2CH2); 69.19 (C-3′); 68.32 Hz, CH2S); 1.98 (s, 3H, CH3CO); 1.68–1.58 (m, 2H,
(C-4″); 67.68 (C-2′); 66.68 (C-5′); 61.47 (C-6″); 59.76 (C-6); SCH2CH2); 1.54–1.43 (m, 2H, OCH2CH2); 1.41–1.21 (m, 4H,
5 5 . 8 4 ( C - 2 ) ; 3 0 . 6 7 , 2 8 . 5 3 , 2 4 . 7 7 , 2 2 . 0 0 OCH2CH2CH2CH2CH2CH2S); 1.12 (d, 3H, J = 6.6 Hz, H-6′.
(OCH2CH2CH2CH2CH2); 22.23 (CH3CO); 15.27 (C-6′); 13.30 13C-NMR (150 MHz, D2O): 174.09 (C=O); 101.83 (C-1″);
(CH2CH3). HRESIMS Calcd for C26H48NO15 [M+H]+ 100.93 (C-1); 98.63 (C-1′); 75.35 (C-5); 74.96, 74.90 (C-3,
614.3024, found 614.3035.
C-5″); 73.41 (C-4); 72.48 (C-3″); 71.91 (C-4′); 71.05 (C-2″);
70.45 (OCH2CH2); 69.22 (C-3′); 68.34 (C-4″); 67.72 (C-2′);
6-Aminohexyl 2-acetamido-2-deoxy-3-O-(α-L- 66.71 (C-5′); 61.48 (C-6″); 59.81 (C-6); 55.86 (C-2); 38.16
fucopyranosyl)-4-O-(β-D-galactopyranosyl)-β-D-gluc- (CH2S); 28.44, 28.30, 27.15, 24.67 (OCH2CH2CH2CH2CH2);
opyranoside (2). The azidotrisaccharide 30 (19 mg, 0.16 mmol) 22.35 (CH3CO); 15.30 (C-6′). HRESIMS Calcd for
was deprotected in the same conditions as described above for C59H92N2O30S2Na [M+Na]+ 1311.5074, found 1311.5065.
the deprotection of trisaccharide 29. After work up, the residue
was passed twice through a Biogel P2 column (100 × 1 cm) 6-Chlorohexyl 2-acetamido-4-O-(2,3,4,6-tetra-O-acetyl-β-D-
eluted with 0.05 M ammonium acetate and after repeated galactopyranosyl)-6-O-benzyl-3-O-(chloroacetyl)-2-deoxy-β-
lyophilization from Milli-Q water (3 × 10 mL) the known [31] D-glucopyranoside (24). BF3·Et2O (150 μL, 1.19 mmol, 2.0
trisaccharide 2 (6.5 mg, 59%) was obtained as the acetate salt in equiv) was added to a solution of the acceptor 4 (300 mg, 0.59
the form of a white amorphous powder. [α]D = −54 (c 0.9, mmol) and glycosyl donor 7 (1.46 g, 2.96 mmol, 5.0 equiv) [39-
H2O), lit. [31]: [α]D = −54.3 (c 1, H2O), 1H NMR (400 MHz, 41] in anhyd CH2Cl2 (15 mL) at 40 °C. The reaction mixture
D2O): δ 5.12 (d, 1H, J = 4.5 Hz, H-1′); 4.83 (m, 1H, H-5′); 4.53 was stirred for 1 h at 40 °C. The reaction was quenched with
(d, 1H, J = 7.5 Hz, H-1); 4.46 (d, 1H, J = 7.5 Hz, H-1″); 4.00 Et3N (170 μL, 1.22 mmol) and the solvent was evaporated.
(dd, 1H, J = 12.0, 1.0 Hz, H-6a); 3.83–3.95 (m, 7H, H-2, H-3, Flash chromatography of the residue (EtOAc–hexanes, 1:1 to
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
6:4) gave the disaccharide 24 (341 mg, 69%) as colorless oil. (PhCH2); 71.34 (C-3); 71.08 (C-5′); 70.67 (C-3′); 69.31
[α]D = −5 (c1.0, CHCl3), 1H NMR (400 MHz, CDCl3): δ (CH2O); 68.73 (C-2′); 68.05 (C-6); 66.81 (C-4′); 61.31 (C-6′);
7.40–7.26 (m, 5H, Ar); 5.72 (d, 1H, J = 9.2 Hz, NH); 5.24 (bd, 57.05 (C-2); 44.99 (CH2Cl); 32.44 (CH2CH2Cl); 29.28
1H, J = 3.4 Hz, H-4′); 5.11 (dd, 1H, J = 10.0, 8.9 Hz, H-3); 4.95 (OCH2CH2); 26.49, 25.19 (OCH2CH2CH2CH2); 23.58, 20.65,
(dd, 1H, J = 10.4, 8.0 Hz, H-2′); 4.78 (dd, 1H, J = 10.4, 3.5 Hz, 20.56, 20.53, 20.47 (CH3CO). HRESIMS Calcd for
H-3′); 4.72 (d, 1H, J = 12.0 Hz, PhCHH); 4.50–4.41 (m, 3H, C35H51ClNO15 [M+H]+ 760.2947, found 760.2928.
H-1, PhCHH); 4.39 (d, 1H, J = 8.0 Hz, H-1′); 4.15–4.01 (m,
4H, H-6a′, H-6b′, ClCH2CO); 4.01–3.88 (m, 2H, H-2, H-4); 6-Chlorohexyl 2-acetamido-4-O-(2,3,4,6-tetra-O-acetyl-β-D-
3.86–3.78 (m, 1H, OCHH); 3.73–3.65 (m, 2H, H-6a, H-6b); galactopyranosyl)-6-O-benzyl-3-O-(2,3,4-tri-O-benzyl-α-L-
3.62 (t, 1H, J = 6.5 Hz, H-5′); 3.53–3.38 (m, 4H, H-5, OCHH, fucopyranosyl)-2-deoxy-β-D-glucopyranoside (29). A solu-
CH2Cl); 2.10, 2.04, 1.93, 1.92 (4 s, 15H, CH3CO); 1.77–1.67 tion of the disaccharide acceptor 28 (100 mg, 0.132 mmol) and
(m, 2H, CH2CH2Cl); 1.61–1.49 (m, 2H, OCH2CH2); 1.44–1.27 fucosyl donor 9 (189 mg, 0.395 mmol, 3.0 equiv) [42] in a mix-
(m, 4H, OCH2CH2CH2CH2). 13C NMR (100 MHz, CDCl3): δ ture of CH2Cl2 and DMF (1:1, 8 mL) containing activated
170.30, 170.16, 169.96, 168.96, 167.34 (C=O); 137.64, 128.57, powdered MS 4Å (400 mg) was stirred at room temp for 30
128.07, 127.97 (Ar); 100.87 (C-1); 100.12 (C-1′); 74.48, 74.40, min. Cu(II)Br2 (88 mg, 0.394 mmol, 3.0 equiv) and Bu4NBr
74.25 (C-3, C-4, C-5); 73.62 (PhCH2); 70.75, 70.60 (C-3′, (131 mg, 0.409 mmol, 3.1 equiv) were added and the reaction
C-5′); 69.27 (CH2O); 69.09 (C-2′); 67.35 (C-6); 66.81 (C-4′); mixture was stirred for 20 h at room temp. The reaction mix-
61.02 (C-6′); 53.45 (C-2); 44.97 (CH2Cl); 40.80 (ClCH2CO); ture was filtered over Celite® and the solids were washed with
32.40 (CH2CH2Cl); 29.21 (OCH2CH2); 26.44, 25.14 CH2Cl2 (5 mL). The filtrate was diluted with CH2Cl2 (60 mL)
(OCH2CH2CH2CH2); 23.25, 20.64, 20.58, 20.48, 20.48 and washed sequentially with brine (50 mL) and saturated aq
(CH3CO). HRESIMS Calcd for C37H52Cl2NO16 [M+H]+ NaHCO3 (6 × 50 mL). The aq layers were re-extracted with
836.2663, found 836.2634.
CH2Cl2 (50 mL) and the combined organic layers were dried
and concentrated. Flash chromatography of the residue
6-Chlorohexyl 2-acetamido-4-O-(2,3,4,6-tetra-O-acetyl-β-D- (EtOAc–hexanes, 3:7) gave the trisaccharide 29 as colorless oil
galactopyranosyl)-6-O-benzyl-2-deoxy-β-D-glucopyranoside (126 mg, 81%). [α]D = −19 (c 0.5, CHCl3), 1H NMR (600
(28). Thiourea (162 mg, 2.13 mmol, 6.0 equiv) was added to a MHz, CDCl3): δ 7.45–7.24 (m, 20H, Ar); 6.02 (bs, 1H, NH);
solution of the disaccharide 24 (298 mg, 0.356 mmol) in a mix- 5.29 (d, 1H, J = 3.2 Hz, H-4″); 5.09 (d, 1H, J = 3.7 Hz, H-1′);
ture of pyridine and EtOH (2:1, 15 mL). The solution was 5.04 (dd, 1H, J = 10.4, 8.2 Hz, H-2″); 4.98 (d, 1H, J = 11.9 Hz,
stirred for 10 h at 70 °C, the solvents removed by evaporation PhCHH); 4.92–4.85 (m, 2H, H-1, PhCHH); 4.85–4.79 (m, 3H,
and the residue co-concentrated with toluene (2 × 10 mL). The H-3″, PhCH2); 4.75 (d, 1H, J = 11.9 Hz, PhCHH); 4.71 (d, 1H,
crude residue was dissolved in CH2Cl2 (20 mL) and washed J = 11.8 Hz, PhCHH); 4.69 (d, 1H, J = 12.0 Hz, PhCHH); 4.58
sequentially with 2 M HCl (10 mL), saturated aq NaHCO3 (10 (d, 1H, J = 8.2 Hz, H-1″); 4.45 (d, 1H, J = 12.0 Hz, PhCHH);
mL) and brine (10 mL). The aq phases were re-extracted with 4.43–4.38 (m, 1H, H-5′); 4.19 (t, 1H, J = 7.6 Hz, H-3);
CH2Cl2 and the combined organic layers were dried and 4.16–4.11 (m, 2H, H-2′, H-6a″), 4.02 (dd, 1H, J = 10.9, 5.9 Hz,
concentrated. Flash chromatography of the residue (EtOAc- H-6b″); 3.97–3.90 (m, 2H, H-4, H-3′); 3.85–3.74 (m, 3H, H-6a,
hexanes, 6:4) gave the pure disaccharide 28 (165 mg, 61%) as a H-6b, OCHHCH2); 3.68 (s, 1H, H-4′); 3.58 (t, 1H, J = 7.0 Hz,
white amorphous powder. [α]D = +1 (c 1.3, CHCl3), 1H NMR H-5″); 3.55–3.45 (m, 4H, H-2, H-5, CH2Cl); 3.44–3.38 (m, 1H,
(400 MHz, CDCl3): δ 7.37–7.26 (m, 5H, Ar); 5.62 (d, 1H, J = OCHHCH2); 2.03, 2.02, 1.98, 1.84, 1.78 (5 s, 15H, CH3CO);
7.7 Hz, NH); 5.32 (bd, 1H, J = 3.4 Hz, H-4′); 5.13 (dd, 1H, J = 1.76–1.69 (m, 2H, CH2CH2Cl); 1.58–1.47 (m, 2H, OCH2CH2);
10.4, 8.0 Hz, H-2′); 4.90 (dd, 1H, J = 10.4, 3.4 Hz, H-3′); 4.74 1.44–1.27 (m, 4H, OCH2CH2CH2CH2); 1.18 (d, 3H, J = 6.5
(d, 1H, J = 8.2 Hz, H-1); 4.68 (d, 1H, J = 12.1 Hz, PhCHH); Hz, H-6′). 13C NMR (150 MHz, CDCl3): δ 170.45, 170.00,
4.47 (d, 1H, J = 12.1 Hz, PhCHH); 4.45 (d, 1H, J = 8.0 Hz, 169.88, 169.85, 169.21 (C=O); 138.72, 138.68, 138.47, 137.78,
H-1′); 4.13–4.05 (m, 2H, H-6a′, H-6b′); 4.04–3.96 (m, 1H, 128.46, 128.40, 128.30, 128.24, 128.11, 127.90, 127.73, 127.69,
H-3); 3.96–3.92 (bs, 1H, OH); 3.90–3.79 (m, 2H, H-5′, OCHH); 127.61, 127.49, 127.28, 127.00 (Ar); 99.38, 99.34 (C-1, C-1″);
3.69–3.57 (m, 3H, H-4, H-6a, H-6b); 3.53–3.41 (m, 4H, H-5, 97.33 (C-1′); 79.82 (C-3′); 76.79 (C-4′); 76.31 (C-2′); 74.23
OCHH, CH2Cl); 3.41–3.31 (m, 1H, H-2); 2.12, 2.03, 1.97, 1.95 (PhCH2); 74.21 (C-5); 74.03 (C-4); 73.58, 73.35 (PhCH2);
(4 s, 15H, CH3CO); 1.78–1.69 (m, 2H, CH2CH2Cl); 1.62–1.50 73.32 (C-3); 72.38 (PhCH2); 70.52 (C-3″); 70.27 (C-5″); 69.23
(m, 2H, OCH2CH2); 1.46–1.29 (m, 4H, OCH2CH2CH2CH2). (OCH2CH2); 68.73 (C-2″); 68.33 (C-6); 66.61 (C-4″); 66.39
13C NMR (100 MHz, CDCl3): δ 170.36, 170.07, 169.98, (C-5′); 60.22 (C-6″); 56.20 (C-2); 44.92 (CH2Cl); 32.38
169.91, 169.15 (C=O); 138.02, 128.48, 127.86, 127.78 (Ar); (CH2 CH2 Cl); 29. 12 (OCH2 CH2 ); 26. 45, 25. 08
101.13 (C-1′); 99.96 (C-1); 80.98 (C-4); 73.92 (C-5); 73.59 (OCH2CH2CH2CH2); 22.95, 20.65, 20.52, 20.49, 20.44
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
(CH3CO). HRESIMS Calcd for C62H79ClNO19 [M+H]+ NMR (400 MHz, CDCl3): δ 7.40–7.20 (m, 20H, Ar); 5.85 (d,
1176.4935, found 1176.4933.
1H, J = 7.7 Hz, NH); 5.25 (d, 1H, J = 3.0 Hz, H-4″); 5.07 (d,
1H, J = 3.8 Hz, H-1′), 5.01 (dd, 1H, J = 10.4, 8.2 Hz, H-2″);
6-Azidohexyl 2-acetamido-4-O-(2,3,4,6-tetra-O-acetyl-β-D- 4.98 (d, 1H, J = 11.8 Hz, PhCHH); 4.90–4.83 (m, 2H, H-1,
galactopyranosyl)-6-O-benzyl-3-O-(2,3,4-tri-O-benzyl-α-L- PhCHH); 4.82–4.74 (m, 3H, H-3″, PhCH2); 4.74–4.62 (m, 3H,
fucopyranosyl)-2-deoxy-β-D-glucopyranoside (30). NaN3 (17 PhCH2, PhCHH); 4.54 (d, 1H, J = 8.2 Hz, H-1″); 4.44–4.35 (m,
mg, 0.26 mmol, 8.2 equiv) was added to a solution of the tri- 2H, H-5′, PhCHH); 4.18 (t, 1H, J = 7.7 Hz, H-3); 4.14–4.05 (m,
saccharide 29 (38 mg, 0.032 mmol) in anhyd DMF (2.5 mL) 2H, H-2′, H-6a″); 3.97 (dd, 1H, J = 10.8, 5.9 Hz, H-6b″);
and the reaction mixture was heated at 80 °C for 36 h. The 3.94–3.85 (m, 2H, H-4, H-3′); 3.83–3.68 (m, 3H, H-6a, H-6b,
solvent was evaporated, the residue was dissolved in CH2Cl2 OCHHCH2); 3.65 (d, 1H, J = 2.7 Hz, H-4′); 3.53 (t, 1H, J = 6.8
(50 mL) and washed with water (2 × 10 mL). The aq phases Hz, H-5″); 3.50–3.44 (m, 1H, H-5); 3.43–3.30 (m, 2H, H-2,
were re-extracted with CH2Cl2 and the combined organic layers OCHHCH2); 2.81 (t, 2H, J = 7.2 Hz, CH2S); 2.29 (s, 3H,
were dried and concentrated. Flash chromatography of the SCOCH3); 1.99, 1.97, 1.93, 1.89, 1.71 (5s, 15H, CH3CO);
residue (EtOAc-hexanes, 6:4) afforded the trisaccharide 30 as a 1.59–1.41 (m, 4H, CH2CH2S, OCH2CH2); 1.33–1.20 (m, 4H,
clear glass (36 mg, 95%). [α]D = −47 (c 1.0, CH2Cl2), 1H NMR OCH2CH2CH2CH2); 1.15 (d, 3H, J = 6.5 Hz, H-6′). 13C NMR
(400 MHz, CDCl3): δ 7.41–7.20 (m, 20H, Ar); 5.81 (d, 1H, J = (100 MHz, CDCl3): δ 195.99, 170.16, 170.06, 169.97, 169.92,
7.6 Hz, NH); 5.26 (d, 1H, J = 3.0 Hz, H-4″); 5.06 (d, 1H, J = 169.19 (C=O); 138.86, 138.80, 138.59, 137.90, 128.53, 128.47,
3.8 Hz, H-1′), 5.00 (dd, 1H, J = 10.4, 8.2 Hz, H-2″); 4.94 (d, 128.37, 128.31, 128.19, 127.97, 127.80, 127.77, 127.67, 127.56,
1H, J = 11.8 Hz, PhCHH); 4.91–4.83 (m, 2H, H-1, PhCHH); 127.34, 127.09 (Ar); 99.45, 99.42 (C-1, C-1″); 97.40 (C-1′);
4.82–4.74 (m, 3H, H-3″, PhCH2); 4.73–4.62 (m, 3H, PhCH2, 79.94 (C-3′); 76.94 (C-4′); 76.40 (C-2′); 74.33 (C-5); 74.31
PhCHH); 4.54 (d, 1H, J = 8.1 Hz, H-1″); 4.43–4.34 (m, 2H, (PhCH2); 74.21 (C-4); 73.66, 73.44 (PhCH2); 73.36 (C-3);
H-5′, PhCHH); 4.20–4.05 (m, 3H, H-3, H-2′, H-6a″); 3.98 (dd, 72.49 (PhCH2); 70.64 (C-3″); 70.34 (C-5″); 69.39 (OCH2CH2);
1H, J = 10.8, 5.9 Hz, H-6b″); 3.95–3.87 (m, 2H, H-4, H-3′); 68.81 (C-2″); 68.40 (C-6); 66.71 (C-4″); 66.40 (C-5′); 60.29
3.83–3.68 (m, 3H, H-6a, H-6b, OCHHCH2); 3.65 (d, 1H, J = (C-6″); 56.60 (C-2); 30.61 (SCOCH3); 29.40, 29.40, 29.20,
1.4 Hz, H-4′); 3.57–3.44 (m, 2H, H-5, H-5″); 3.43–3.31 (m, 2H, 28.96, 28.43, 25.36 (OCH2CH2CH2CH2CH2CH2S); 23.18,
H-2, OCHHCH2); 3.20 (t, 2H, J = 6.9 Hz, CH2N3); 1.99, 1.98, 20.70, 20.59, 20.56, 20.51 (CH3CO); 16.71 (C-6′). HRESIMS
1.93, 1.89, 1.70 (5s, 15H, CH3CO); 1.58–1.43 (m, 4H, Calcd for C64H82NO20S [M+H]+ 1216.5151, found 1216.5151.
C H 2 C H 2 N 3 , O C H 2 C H 2 ) ; 1 . 3 3 – 1 . 2 1 ( m , 4 H ,
OCH2CH2CH2CH2); 1.15 (d, 3H, J = 6.5 Hz, H-6′). 13C NMR 6-Benzylthiohexyl 2-acetamido-4-O-(2,3,4,6-tetra-O-acetyl-
(100 MHz, CDCl3): δ 170.09, 170.06, 169.96, 169.91, 169.20 β-D-galactopyranosyl)-6-O-benzyl-3-O-(2,3,4-tri-O-benzyl-
(C=O); 138.88, 138.77, 138.57, 137.89, 128.53, 128.47, 128.38, α-L-fucopyranosyl)-2-deoxy-β-D-glucopyranoside (32).
128.32, 128.19, 127.97, 127.77, 127.75, 127.66, 127.56, 127.36, PhCH2SH (60 µL, 0.44 mmol, 15 equiv) and NaH (21 mg, 0.44
127.08 (Ar); 99.45 (C-1, C-1″); 97.44 (C-1′); 79.97 (C-3′); mmol, 15 equiv) were added to a solution of the trisaccharide
76.90 (C-4′); 76.42 (C-2′); 74.19 (C-5, C-4); 73.70 (PhCH2); 29 (36 mg, 0.030 mmol) in anhyd DMF (3.0 mL) at room temp.
73.43 (C-3, PhCH2); 72.46 (PhCH2); 70.62 (C-3″); 70.35 After 10 min the reaction mixture was heated to 80 °C for 16 h,
(C-5″); 69.29 (OCH2CH2); 68.80 (C-2″); 68.43 (C-6); 66.69 the solvent was evaporated and the residue was dissolved in
(C-4″); 66.42 (C-5′); 60.28 (C-6″); 56.60 (C-2); 51.32 (CH2N3); Ac2O and pyridine (5 ml, 1:1). After 18 h the reaction mixture
29.23, 28.73, 26.41, 25.43 (OCH2CH2CH2CH2CH2CH2N3); was co-concentrated with toluene (3 × 20 ml), the residue was
23.16, 20.70, 20.59, 20.56, 20.51 (CH3CO); 16.71 (C-6′). dissolved in CH2Cl2 (30 mL) and the solution was washed with
HRESIMS Calcd for C62H79N4O19 [M+H]+ 1183.5339, found water (2 × 10 mL). The aq phases were re-extracted with
1183.5325.
CH2Cl2 and the combined organic layers were dried and
concentrated. Flash chromatography of the residue
6-Acetylthiohexyl 2-acetamido-4-O-(2,3,4,6-tetra-O-acetyl-β- (EtOAc–hexanes, 1:1) gave the trisaccharide 32 (35.6 mg, 94%)
D-galactopyranosyl)-6-O-benzyl-3-O-(2,3,4-tri-O-benzyl-α- as a white solid. [α]D = −28 (c 1.0, CH2Cl2), 1H NMR (400
L-fucopyranosyl)-2-deoxy-β-D-glucopyranoside (31). MHz, CDCl3): δ 7.44–7.13 (m, 25H, Ar); 5.79 (d, 1H, J = 7.6
KSC(O)CH3 (26 mg, 0.22 mmol, 10 equiv) was added to a Hz, NH); 5.25 (d, 1H, J = 3.0 Hz, H-4″); 5.05 (d, 1H, J = 3.8
solution of the trisaccharide 29 (27 mg, 0.023 mmol) in anhyd Hz, H-1′), 5.00 (dd, 1H, J = 10.5, 8.2 Hz, H-2″); 4.94 (d, 1H, J
DMF (1.5 mL) and the reaction mixture was heated at 80 °C for = 11.8 Hz, PhCHH); 4.90–4.74 (m, 5H, H-1, H-3″, PhCH2,
16 h. Work up and chromatography (EtOAc–hexanes, 6:4), as PhCHH); 4.73–4.64 (m, 3H, PhCHH, PhCH2); 4.54 (d, 1H, J =
described above for compound 30 gave the trisaccharide 31 as 8.2 Hz, H-1″); 4.43–4.36 (m, 2H, H-5′, PhCHH); 4.17 (t, 1H, J
colorless glass (19 mg, 70%). [α]D = −43 (c 0.7, CH2Cl2), 1H = 7.7 Hz, H-3); 4.14–4.06 (m, 2H, H-2′, H-6a″); 4.01–3.95 (m,
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Beilstein Journal of Organic Chemistry 2010, 6, No. 17.
5. Nakasaki, H.; Mitomi, T.; Noto, T.; Ogoshi, K.; Hanaue, H.; Tanaka, Y.;
Makuuchi, H.; Clausen, H.; Hakomori, S.-I. Cancer Res. 1989, 49,
3662–3669.
1H, H-6b″); 3.93–3.87 (m, 2H, H-4, H-3′); 3.79–3.64 (m, 6H,
H-6a, H-6b, H-4′, SCH2Ph, OCHHCH2); 3.56–3.45 (m, 2H,
H-5, H-5″); 3.41–3.31 (m, 2H, H-2, OCHHCH2); 2.36 (t, 2H, J
= 7.3 Hz, CH2SBn); 1.99, 1.98, 1.94, 1.90, 1.70 (5s, 15H,
CH3CO); 1.52–1.42 (m, 4H, CH2CH2S, OCH2CH2); 1.33–1.28
(m, 4H, OCH2CH2CH2CH2); 1.15 (d, 3H, J = 6.4 Hz, H-6′).
13C NMR (150 MHz, CDCl3): δ 170.14, 170.10, 170.02,
169.96, 169.20 (C=O); 140.06, 138.91, 138.81, 138.61, 137.93,
137.63, 130.14, 129.02, 128.97, 128.53, 128.49, 128.30, 128.00,
127.89, 127.73, 127.60, 127.39, 126.89 (Ar); 99.48, 99.41 (C-1,
C-1″); 97.47 (C-1′); 80.05 (C-3′); 76.86 (C-4′); 76.42 (C-2′);
74.34 (C-5); 74.31 (PhCH2); 74.25 (C-4); 73.77, 73.46
(PhCH2); 73.35 (C-3); 72.47 (PhCH2); 70.67 (C-3″); 70.33
(C-5″); 69.49 (OCH2CH2); 68.81 (C-2″); 68.39 (C-6); 66.70
(C-4″); 66.41 (C-5′); 60.28 (C-6″); 52.07 (C-2); 36.30
(S-CH2Ph); 31.30 (CH2SBn); 29.29, 29.13, 29.06, 25.53
(OCH2CH2CH2CH2CH2CH2S); 23.23, 20.75, 20.64, 20.61,
20.57 (CH3CO); 16.76 (C-6′). HRESIMS Calcd for
C69H86NO19S [M+H]+ 1264.5515, found 1264.5509.
6. Singhal, A. K.; Ørntoft, T. F.; Nudelman, E.; Nance, S.; Schibig, L.;
Stroud, M. R.; Clausen, H.; Hakomori, S.-I. Cancer Res. 1990, 50,
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0
10.Satoh, J.; Kim, S. U. J. Neurosci. Res. 1994, 37, 466–474.
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11.Croce, M. V.; Isla-Larrain, M.; Rabassa, M. E.; Demichelis, S.;
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doi:10.1016/0008-6215(92)84057-Y
Supporting Information
Supporting Information File 1
Experimental procedures and characteristics for compounds
4–6, 8, 11, 12, 14–19, 21–23, 25–27.
16.Numomura, S.; Iida, M.; Numata, M.; Sugimoto, M.; Ogawa, T.
Carbohydr. Res. 1994, 263, C1–C6.
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[http://www.beilstein-journals.org/bjoc/content/
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1H and 13C NMR spectra for compounds 1–6, 8, 11, 12,
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Acknowledgements
We thank the National Science and Engineering Research
Council of Canada, the Canada Foundation for Innovation and
the Ontario Innovation Trust for financial support of this work.
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