Structure-Reactivity Relationships of Conformationally Armed Disaccharide Donors and Their Use in the Synthesis of a Hexasaccharide Related to the Capsular Polysaccharide from Streptococcus pneumoniae Type 37

Article abstract of DOI:10.1021/acs.joc.7b01264

To advance the field of glycobiology, efficient synthesis methods of oligosaccharides and glycoconjugates are a requisite. In glycosylation reactions using superarmed donors, both selectivity and reactivity issues must be considered, and we herein investigate these aspects for differently protected β-linked 2-O-glycosylated glucosyl donors carrying bulky tert-butyldimethylsilyl groups to different extents. The acceptors in reactions being secondary alcohols presents a challenging situation with respect to steric crowding. Conformational pyranose ring equilibria of the superarmed disaccharide donors with axial-rich substituents contained skew and boat conformations, and three-state models were generally assumed. With NIS/TfOH as the promotor, 2,6-di-tert-butyl-4-methylpyridine as the base, and a dichloromethane/toluene solvent mixture, ethyl 1-thio-β-d-glucosyl disaccharide donors having 6-O-benzyl group(s) besides tert-butyldimethylsilyl groups were efficiently coupled at -40 °C to the hydroxyl group at position 3 of glucopyranosyl acceptors to form β-(1 → 2),β-(1 → 3)-linked trisaccharides, isolated in excellent 95% yield. The more axial-rich donors in skew and boat conformations are thus preorganized closer to the assumed transition state in these glycosylation reactions. The developed methodology was subsequently applied in the synthesis of a multibranched hexasaccharide related to the capsular polysaccharide from Streptococcus pneumoniae type 37, which consists of a β-(1 → 3)-linked backbone and a β-(1 → 2)-linked side chain of d-glucosyl residues in disaccharide repeating units.

Full text of DOI:10.1021/acs.joc.7b01264

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Article
Structure-Reactivity Relationships of Conformationally Armed Disaccharide
Donors and Their Use in Synthesis of a Hexasaccharide Related to
the Capsular Polysaccharide from Streptococcus pneumoniae type 37
Thibault Angles d'Ortoli, Christoffer Hamark, and Göran Widmalm
J. Org. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.joc.7b01264 • Publication Date (Web): 06 Jul 2017
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Structure-Reactivity Relationships of Conformationally Armed Disaccharide
Donors and Their Use in Synthesis of a Hexasaccharide Related to the Capsular
Polysaccharide from Streptococcus pneumoniae type 37
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Thibault Angles d’Ortoli§, Christoffer Hamark§ and Göran Widmalm*
Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University,
S-106 91 Stockholm, Sweden
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TOC GRAPHIC
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ABSTRACT
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To advance the field of glycobiology efficient synthesis methods of oligosaccharides and
glycoconjugates are a requisite. In glycosylation reactions using super-armed donors both
selectivity and reactivity issues must be considered, and we herein investigate these aspects for
differently protected β-linked 2-O-glycosylated glucosyl donors carrying bulky tert-
butyldimethylsilyl groups to different extent. The acceptors in reactions being secondary alcohols
present a challenging situation with respect to steric crowding. Conformational pyranose ring
equilibria of the super-armed disaccharide donors with axial-rich substituents contained skew and
boat conformations and three-state models were generally assumed. With NIS/TfOH as promotor,
2,6-di-tert-butyl-4-methylpyridine as base and a dichloromethane:toluene solvent mixture ethyl 1-
thio-β-D-glucosyl disaccharide donors having 6-O-benzyl group(s) besides tert-butyldimethylsilyl
groups were efficiently coupled at –40 °C to the hydroxyl group at position three of glucopyranosyl
acceptors to form β-(1→2),β-(1→3)-linked trisaccharides, isolated in excellent 95% yield. The
more axial-rich donors in skew and boat conformations are thus pre-organized closer to the assumed
transition state in these glycosylation reactions. The developed methodology was subsequently
applied in synthesis of a multi-branched hexasaccharide related to the capsular polysaccharide from
Streptococcus pneumoniae type 37, which consists of a β-(1→3)-linked backbone and a β-(1→2)-
linked side-chain of D-glucosyl residues in disaccharide repeating units.
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KEY WORDS: super-armed donors, convergent carbohydrate synthesis, glycan building-blocks,
preorganization, conformational analysis, NMR spin-simulation, transition state conformation, ring
puckering
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INTRODUCTION
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In the present era of functional glycomics, light is increasingly shed on the vast number of
important functions of carbohydrates in biological processes.1–3 Glycans act as receptors for, inter
alia, cells, hormones and pathogens and they govern immune reactions. This is valid to the extent
that there is essentially no vital process in mammalians, including pathogenicity, not intervened by
carbohydrates.4 The advancement of efficient protocols for synthesis of these molecules is therefore
of great importance. Chemical synthesis in general, and carbohydrate synthesis in particular, require
control over selectivity, protective group orthogonality and substrate reactivity.5–9 Due to their
manifestation of numerous hydroxyl groups and the special nature of the anomeric center, the
preparation of homologues oligosaccharides with minor structural modifications generally involve
multi-step procedures. The herein presented study aims at systematically introducing functional
glycan building blocks for the application in convergent syntheses of challenging targets.
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2-O-Glycosylated glucosyl donors introduce selectivity issues as well as steric bulk, impeding
acceptor accessibility in glycosylation reactions.10 If a participating group is not present at O2 and if
the β-anomeric configuration is desired, other means of directing selectivity have to be employed.
Yamada and co-workers have proposed the introduction of bulky protective groups, particularly at
position 2 of an aldohexose, to induce a ring-flip into a skew conformation and thus sterically
hinder access to the α-face of the donor.11 This approach resulted in excellent selectivity but
moderate yields for various targets (Scheme 1a).12 Changes in ring conformation into axial-rich
arrangements bring about additional benefits to glycosyl donors, i.e., the reactivity increases. A
decade ago, Bols and co-workers introduced the notion of conformational arming,13,14 thereby
extending the established armed-disarmed concept15 to comprise super-armed glycosyl donors.16 In
subsequent studies various 1-thio-glycoside donors were investigated, showing a 20-fold reactivity
enhancement17 (Scheme 1b) and more recently a 100-fold higher reactivity18 compared to
non-axial-rich analogues. Conformationally super-armed thio-ethyl glycosyl donors have also been
used in one-pot synthesis of trisaccharides.19 Furthermore, for alkoxy substituents, axial
arrangements will be less destabilizing for the charged transition state through an amplified charge-
dipole interaction.20,21 Coupling reactions with 2-O-glucosylated glucosyl donors are intrinsically
challenging due to potential steric crowding as was noted in the synthesis of the four anomeric
combinations of D-Glcp-(1→2)-D-Glcp-(1→3)-α-D-Glcp-OMe,10 a structural arrangement that we
will refer to as (1→2),(1→3)-linked trisaccharides. In targeting tetra- and pentasaccharides
containing β-D-Glcp residues at these types of linkages, glucosyl monosaccharide donors were
employed.22
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Scheme 1. Examples of axial-rich donors with the purposes of attaining stereoselectivity (a),
enhanced reactivity (b), or stereoselectivity as well as enhanced reactivity (c) by work of Yamada
and co-workers, Bols and co-workers, and Angles d’Ortoli and Widmalm, respectively.12,13,23
To utilize disaccharides efficiently in such syntheses very potent donors would be necessary. We
recently showed that by employing a super-armed donor, glycosylation of a disaccharide acceptor
was possible (Scheme 1c) resulting, after deprotection, in the tetrasaccharide glycoside moiety of
the glycoalkaloid Solaradixine.23 By using different super-armed disaccharide donors and exploring
their scope in glycosylation reactions we herein show that not only can excellent selectivity in the
formation of β-linked trisaccharides products11 be obtained but also excellent yields for challenging
situations, in which the hydroxyl group is attached to a secondary carbon atom. Tetrasaccharides
were also synthesized from the same donors using a convergent approach employing a disaccharide
acceptor, and the reaction conditions were investigated to shed light on the influence of steric
crowding. With this acquired knowledge we apply the developed methodology in the synthesis of a
hexasaccharide (Figure 1) corresponding to three repeating units of the capsular polysaccharide
(CPS) from Streptococcus pneumoniae type 37 and the exopolysaccharide from Propionibacterium
freudenreichii ssp. shermanii JS.24,25
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Figure 1. Schematic of a glucosyl-containing hexasaccharide (14) corresponding to three repeating
units of the CPS from S. pneumoniae type 37. The oligosaccharide is colored to highlight its
repetitive disaccharide structural elements. The sugar residues are denoted by capital letters in the
β-(1→3)-linked backbone and by primed capital letters in the β-(1→2)-linked side-chains.
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RESULTS AND DISCUSSION
Synthesis of glycosyl donors.
Synthesis of armed donors D1, D3 and D5 (Scheme 2) having a 2-naphthylmethyl (NAP or just
naphthyl) group26 at O3 of the reducing end residue started from previously described disaccharides
1, 4 and 6.23 Acetal 1 was hydrolyzed at 80 °C for 2 h27 using a solution of 80% AcOH (aq), which
gave diol 2 in 82% yield. Compounds 2, 4 and 6 had their O-acyl groups removed under standard
conditions employing a solution of 1 M NaOMe in MeOH and the resulting hexaol 3, pentaol 5 and
tetraol 7 were obtained in 90 − 92% yield. In separate experiments, alcohols 3, 5 and 7,
respectively, together with 4-dimethylaminopyridine (DMAP), were dissolved in pyridine;
TBDMSOTf was added at 0 °C and the mixture was stirred overnight at 80 °C. Fully protected
super-armed donors D1, D3 and D5 were isolated after flash column chromatography in 85 – 88%
yield. The syntheses of the super-armed donors D2, D4 and D6 have been described previously.11,23
In total, six donors, D1 – D6, armed to different extent (Figure 2a) are thus available for subsequent
glycosylation reactions.
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Scheme 2. Synthesis of armed donors D1, D3 and D5. Reagents and conditions: (a) 80% AcOH
(aq), 80 °C, 4 h, 82%; (b) 1 M NaOMe, MeOH, 16 h, 90 – 92%; (c) TBDMSOTf, DMAP, Pyr, 80
°C, 24 h, 85 − 88%.
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Figure 2. Disaccharide donors (a) and acceptors (b) used to investigate reactivity in glycosylation
reactions and isolated by-products (c).
Conformational analysis of glycosyl donors.
The differently functionalized β-(1→2)-linked disaccharide donors D1 – D6 all have tert-
butyldimethylsilyl substituents (TBS), which are bulky protective groups known to induce
distortions to the pyranose chair conformation (Scheme 1a). It has been shown that TBS-group
1
insertion at positions 2 and 3 on β-D-glucopyranose forces the ring into a C4 conformation,28
whereas the same substituents at positions 3 and 4 do not change the ring puckering; to alter the ring
conformation in the latter case requires an even bulkier protecting group, viz., tert-
butyldiphenylsilyl (TBDPS) group at O3 and at O4.29 Furthermore, full TBS-protection yields a 3S1
ring conformation in β-D-glucopyranoside donors12,14,30 and this conformation has also been
observed for 2-O-glycosylated12 (Scheme 1a) and 6-O-benzylated12,14 (Scheme 1b) derivatives of 1-
thio-β-D-glucose.
To this end we set out to investigate how various types of protective groups would affect the
conformation and consequently the reactivity of 1-thio-β-D-glucosyl donors. The introduction of a
NAP group, orthogonal to silyl and benzyl ethers as well as to ester protective groups, would enable
selective deprotection. It is known that glycosylation with an axial-rich donor equipped with an acid
labile group at position 6 at the reducing end (e.g. D1 and D2 in Figure 2a) could afford internally
cyclized by-products, viz. 1,6-anhydro-hexopyranose derivatives.14 The exchange for a more robust
protective group, such as a benzyl ether, has shown to prevent this by-product formation14 and such
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a group was introduced in compounds D3 – D6. Given the fact that the terminal glucose units of the
donors also reside in distorted conformations, computer modeling suggested that protective groups
at O6 of these residues, may explore three-dimensional space to different extent (Figure S1). The
insertion of a benzyl group, instead of a TBS group, might thus impede steric hindrance, as in D5
and D6.
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By means of NMR spectroscopy, the donor ring protons were exploited to relate their J couplings to
torsion angles and thereby ring conformations by Karplus-type relationships. Initially, H
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resonances were assigned using 1D and 2D NMR experiments suitable for carbohydrates31 and the
chemical shifts and scalar coupling constants were refined by NMR spin-simulation using an
iterative total-lineshape analysis approach (Figure S2 and Table S1).32 Observed J couplings will
reflect population-weighted averages and by fitting experimental data to models, a distribution can
be obtained. Built structures of relevant canonical ring puckers33 were energy minimized and with
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the generalized Haasnoot-Altona equation34 their ring-defining JHH couplings were calculated.
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These computed JHH values were compared by a root-mean-square deviation (RMSD) to those
experimentally determined. The RMSD was minimized with the generalized reducing gradient
algorithm,35 by altering the populations of the different conformations and a weighted population
distribution was thus achieved for both residues of donors D1 – D6 (Table 1a). For the residues
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populating axial-rich conformations, significant long-range JHH couplings were observed for the
H2-H4 and H3-H5 proton pairs typically being around −1 Hz and −0.7 Hz, respectively (Table S1),
which supported the presence of the conformational equilibria established. It should be noted that
the conformational equilibria for each donor correspond to similar or adjacent conformers with
puckering geometries that readily interconvert in-between each other for itineraries on the
conformational sphere as described for β-D-glucose by Mayes et al.33
Table 1. Populations (%) of donors D1 – D6 (a) and products P3b and P4 (b) at 25 °C in CDCl3.
a
Residue B'
Residue B
1S5
3S1 RMSD 4C1 3S1 1C4 RMSD 1S5
3S1 RMSD
3,OB
3,OB
D1
D2
D3
D4
D5
D6
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0
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84 12
85 13
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28 72
13 87
11 84
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37 57
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b
Residue B'
Residue B
Residue A
4C1 3S1 1C4 RMSD 4C1 1S5 1C4 RMSD 4C1 RMSD
0
0
73 27
70 29
0.09
0.08
12 88
94
0
3
0.19 100 0.25
0.17 100 0.23
P3b
P4
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The results of the conformational analysis indicated interconversion pathways somewhat extending
from the geometry itineraries of cyclohexane proposed by Stoddart36 to include transformations
between stable energy minimum puckers for β-glucose (Scheme 3), investigated by Beckham and
co-workers in a recent study.33 The terminal residues (B' in Table 1a), not directly affecting the
reactivity, were all shown to reside in a major 3S1 conformation as would be expected from previous
studies (vide supra). However, this single conformer was not sufficient to describe the
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conformational space and an equilibrium with the axial-rich arrangements S1 ⇌ 3,OB ⇌ S5 was
considered. Upon benzylation at the O6 position, as in D5 and D6, a slight shift toward a higher
population of 3,OB was observed. Conformational changes were also investigated by analysis of
3JH5,H6 coupling constants for the terminal residue B', which in D4 had JH5,H6pro-R = 5.63 Hz and
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3JH5,H6pro-S = 8.15 Hz; in D6 these were JH5,H6pro-R = 6.30 Hz and JH5,H6pro-S = 6.34 Hz. Based on
these spin-spin coupling constants the rotamer populations at the ω torsion angle were estimated.37
In D4 a relative distribution of 0.24:0.04:0.72 was deduced for the gt:gg:tg conformational states
and in D6 the corresponding distribution was 0.39:0.09:0.52. Given the fact that the J couplings and
thus rotamer distribution do differ, besides the 3D shape of the protecting group per se, these data
suggest an increased flexibility upon exchanging the TBS group for the benzyl group. The
corresponding effect was not observed for D3 vs D5 (cf. Figure S1). For these compounds the inter-
residual strain is likely to be lower due to the difference in conformations between the two rings.
3
3
3
Thus, each conformational equilibrium of donors in Table 1 is present with a major S1 skew
conformation and adjacent ones that are populated via interconversion between states.
Scheme 3. Selected ring conformational interconversion pathways of β-D-glucose adapted from
Beckham and co-workers,33 including local minima and transition states between them. Only the
pyranoid ring is shown for simplicity.
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For the reducing end moieties (B in Table 1a) it is evident that donors with a large naphthyl group
at O3 (D1, D3 and D5) instead of a TBS group, do not undergo any major interconversion from 4C1,
despite the bulkiness of the bicyclic aromatic group. Nevertheless, a slight perturbation of the
equilibrium, involving also the S1 conformer, was detected; changes in conformation upon
benzylation at O6 were not detected. For the reducing end residue of D2, D4 and D6, a more equal
3
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population distribution along the pathway 3S1 ⇌ 3,OB ⇌ S5 was observed, where benzylation of O6
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(D4 and D6) shifted the equilibrium toward the S5 conformer. Since the subsequent glycosylation
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reactions were all performed at temperatures between –80 and –20 °C, an investigation of the H
NMR temperature dependence was conducted for donor D4 at temperatures between –30 and 25 °C
(Figure S3). The results indicated that residue B' does not change conformation with temperature
whereas for residue B the conformational equilibrium shifts as a consequence of temperature
changes. The resonances of residue B are successively broadened as a result of lower temperature,
further supporting the presence of a dynamic conformational equilibrium.
Glycosylations.
Trisaccharides. – The glycosylation reactions were first investigated using donors D1 and D2; it
was anticipated that product formation would be difficult since previous attempts with similar but
monosaccharide donor substrates only gave 1,6-anhydro products.14 As a matter of fact, using
different kind of promoter systems, several attempts were made to couple donors D1 and D2 with
acceptor A1 (Figure 2b), which all resulted in the formation of the intramolecularly glycosylated
by-products B1 and B2 (Figure 2c). The use of reaction conditions developed by Okada et al.11,12 to
couple donor D1 and acceptor A1, MeOTf and 2,6-lutidine as base in DCM as such, only resulted to
traces amount of trisaccharide product P1 with the major formation of compound B2. It is to be
noted that 1,6-anhydro by-product formation was higher employing donor D2 as opposed to D1,
65% and 48%, respectively, with NIS/TfOH as the activator system (Scheme 4). From these results
it was concluded that D2 is more reactive than D1, which carries a NAP group at O3. Nevertheless,
the latter compound still undergoes the internal cyclization, which is a strong indication that it
readily experiences a ring-flip toward an axial-rich conformation, which the conformational
analysis revealed (Table 1a).
Subsequently, suitable glycosylation procedures for donors D3 – D6 were elaborated. Donor D4
represented the potentially most acid labile compound and was therefore selected as a model for
optimizations of the reaction conditions in glycosylations with acceptor A2 (Figure 2b). At an
activation temperature of –78 °C, a wide range of relevant promoter systems were tested (Table 2)
including MeOTf and NIS/TfOH, previously described to be suitable for related reactions.11,12,14
NIS/TfOH appeared to give the best results and gave rise to the least by-product formation; it was
consequently chosen in further optimizations. The reactions were all monitored by MS analyses,
which revealed that cleavage of the silyl ether protective groups were major side-reactions. Also
hydrolysis reactions were repeatedly observed. To minimize the loss of the acid-labile silyl ethers,
different sterically hindered bases were tested during the glycosylation reactions. The bulky base
2,6-di-tert-butyl-4-methylpyridine (DTBMP) gave the best results for neutralizing acidic species in
the reaction solution. Interestingly, it was only when dry toluene was added in equal proportions to
dry dichloromethane, that hydrolysis reactions could be suppressed yielding the desired product as
the major one. Allowing the temperature to reach –40 °C and employing the optimized conditions
permitted isolation of trisaccharide P4 in an excellent 95% yield (Scheme 4).
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Table 2. Glycosylation of D4 with A2.
Activation
temp.
Amount
By-products
Entry
Activator
Base
Solvent
Yield (%)
TfO-/Tf2O
i:ii (%)a
(°C)
6
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9
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5
6
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9
MeOTf
NIS/AgOTf
NIS/TMSOTf
NIS/TfOH
Tf2O-DMDS
Tf2O-DPSO
Tf2O-BSP
NIS/TfOH
NIS/TfOH
2,6-Lutidine
2,6-Lutidine
2,6-Lutidine
2,6-Lutidine
DTBMP
TTBP
TTBP
DTBMP
DTBMP
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
traces
25
20
25
29
17
20
40
95
major:minor
minor:major
minor:major
minor:major
major:minor
minor:51
minor:48
minor:55
traces:traces
cat
cat
cat
−78 → RT
−78
−78
−78
−78
−78
−78
−78
cat
1.1 eq
1.25 eq
1.25 eq
cat
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
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37
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40
41
42
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44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
CH2Cl2/Tol 1:1
cat
−78 → −40
a i and ii refer to hydrolysis- or elimination by-products and de-O-silylated by-products,
respectively. Entries devoid of values infer by-product detected with MS, otherwise isolated yield.
DMDS: Dimethyl disulfide;
DPSO: Diphenyl sulfoxide;
BSP: Benzenesulfinylpiperidine;
DTBMP: 2,6-Di-tert-butyl-4-methylpyridine;
TTBP: 2,4,6-Tri-tert-butylpyrimidine.
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Scheme 4. Glycosylation reactions using different donors and acceptors, products formed and the
respective yields. Reagents and conditions: (a) NIS, TfOH, 4 Å MS, DCM. (b) NIS, TfOH,
DTBMP, 4 Å MS, DCM/Tol 1:1.
The subsequent glycosylations with donors D3 – D6 were performed using the improved conditions
stated by entry 9 in Table 2 though the reactions temperature was set to −60 ºC to increase solubility
of the acceptors (A1 and A3 in particular) in the toluene-containing solvent mixture. These changes
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had limited or no effect on reaction yields. Donor D6 was reacted with acceptor A2 to give product
P6 in 95% yield also (Scheme 4). NAP-protected donor D3 was reacted with A1 and A2 yielding
products P3a and P3b, respectively, in moderate to good isolated yields. Compound P3b was
obtained in a better yield than for P3a due to the fact that A2 is a more electron-rich nucleophile
than acceptor A1, but steric aspects may also influence the yield. Furthermore, donor D5 was
coupled with acceptors A2 whereafter product P5b was isolated in a good yield of 75%.
10
11
12
13
14
15
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In the previous synthesis of a β-(1→2),β-(1→3)-linked glucosyl trisaccharide (cf. compounds P4,
P5b and P6) a disaccharide acceptor corresponding to residues B and A was used employing a
monosaccharide donor, corresponding to residue A', in the silver triflate promoted glycosylation
reaction to furnish the target trisaccharide in a good 75% yield.10 These glycosylations utilized
monosaccharide donors thereby ‘capping’ the acceptor in syntheses whose directionality can be
described as ‘toward the nonreducing end’. The developments of the super-armed disaccharide
donors facilitate chemical synthesis in a direction ‘toward the reducing end’ of an oligosaccharide.
Importantly, and in contrast to our previous synthesis protocols, we have herein shown that it is
possible to carry out glycosidic bond formation with complete stereoselectivity in an excellent yield
of 95% using a disaccharide donor and an acceptor where the substitution takes place at a
secondary alcohol, i.e., resulting in that the C3 atom of a glucose residue becomes glycosyloxylated
in the present case.
Tetrasaccharides. – The highly O-acetylated, disarmed, as well as sterically crowded acceptor A3
(Figure 2b), which was synthesized in a two-step procedure by glycosylation of 2,3,4,6-tetra-O-
acetyl-β-D-glucopyranosyl
trichloroacetimidate
with
methyl
3-p-methoxybenzyl-4,6-O-
benzylidene-α-D-glucopyranoside followed by removal of the p-methoxybenzyl group by DDQ,
was coupled with donor D3 in a 2+2 fashion,23,38 to form tetrasaccharide P3c, but only in 30%
yield. Additionally, donor D5 was coupled with acceptor A3, whereafter product P5c was isolated
in a moderate 56% yield. All glycosylation reactions gave excellent anomeric selectivity for the
formation of the β-(1→3)-linkages as would be expected from the work of Yamada and co-
workers.11 (vide supra). Indeed, none or only supposedly trace amounts of oligosaccharides having
1
the α-configuration at this glycosidic linkage could be observed in H NMR spectra of isolated
products. In the earlier synthesis of the tetrasaccharide corresponding to compound P5c a β-(1→3)-
linked disaccharide acceptor was used and condensed with two monosaccharides as donors to
produce the tetrasaccharide in a very good yield of 84%.22
Conformational aspects related to product formation.
The results from the glycosylations clearly show that the conformationally distorted D2, D4 and D6
donors were significantly more reactive compared with D1, D3 and D5. The still reasonable yields
when donors D3 and D5 were used, given the hindered system, may be explained by the existence
3
of a minor population of the S1 conformation for the reacting residues, thus implying a slight
conformational arming effect (Table 1a). This reasoning is supported by the substantial
intramolecular by-product formation to B1 over an intermolecular reaction when glycosylation was
carried out with D1, not likely to occur under the employed conditions if the donor was not in an
axial-rich conformation. Also, two of the products, namely trisaccharides P3b and P4 were
subjected to conformational analyses (Table 1b) and clearly, the increased level of steric bulk had
an impact on the conformational preferences. Interestingly, both these products showed similar
conformations where all three residues had distinctly different shapes. The terminal residue B' was
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still adopting mainly a S1 conformation but now in equilibrium with the C4 chair conformation.
Residue B populates mainly a 1S5 conformation, also for P3b, bearing a less bulky naphthyl group.
4
1
The conformationally constrained residue A resided in the expected C1 chair conformation. H
NMR spectra and resonance assignments of products P5b and P6 showed that they were highly
comparable to those of P3b and P4, thus indicating the same type of conformational behavior of all
isolated trisaccharides products.
9
10
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The glycosylation reactions of the type studied herein have been proposed to proceed via an SN1-
like mechanism with an oxocarbenium like transition state (TS).12 Thus, the bulky nature of the
protective groups in the glycosyl donors D1 – D6 imply that axial arrangements of exocyclic
substituents on the pyranose sugar ring should be adopted also in the transient oxocarbenium
intermediate. The influence of conformational preorganization could constitute an additional basis
for the reactivity enhancements of D2, D4 and D6 as well as similar conformationally armed
donors. The pyranose rings of these compounds have puckered to adopt conformations on the
equator of the pseudorotational hemisphere33 and the energy penalty for leaving the low energy well
of the 4C1 conformation is already paid. Consequently, the prearranged axially oriented dipoles can
stabilize the buildup of positive charge in the course of oxocarbenium ion formation. Donors D1,
D3 and D5 reside mainly in C1 conformations but the small presence of S1 might be sufficient to
drive the reaction via this conformer and therefore proceed via a charge-dipole stabilized TS, which
is supported by the significant 1,6-anhydro formation (B1) when D1 is used. The difference in
reactivity compared to D2, D4 and D6 may thus be explained by the altered level of conformational
preorganization.
4
3
Synthesis of a multi-branched hexasaccharide and comparison to polysaccharide NMR data.
In order to avoid potential and superfluous steric clashes during the 2+4 glycosylation step, it was
decided to first remove all bulky silyl ether groups on compound P5c (Scheme 5) and subsequently
replace them by smaller O-acetyl protective groups. A 1 M solution of tetrabutylammonium
fluoride (TBAF) in tetrahydrofuran (THF) was employed to cleave tert-butyldimethylsilyl ether
groups and then directly after a simple workup, the obtained tetraol 7 was O-acetylated using acetic
anhydride (Ac2O) in pyridine in presence of a catalytic amount of DMAP. Thus, compound 8 was
obtained in 92% yield over two steps. An excess of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone
(DDQ) in the solvent mixture DCM/MeOH 4:1 removed the NAP protecting group as well as the
labile benzylidene acetal and triol 9 was furnished in 85% yield. Subsequently, position 6 on sugar
residue
A
was selectively O-silylated39 at −20 ºC using tert-butyldimethylsilyl
trifluoromethanesulfonate (TBDMSOTf) and 2,6-lutidine in DCM to form diol acceptor 10 in 95%
yield. The hydroxyl group at position 4 of residue A was judged to be less reactive than that at
position 3 of residue B in 10 and a glycosylation that should be selective was therefore tried out.
Sugars were dissolved as described above in a mixture of dry DCM/Tol 1:1; reagents and reaction
temperature employed were the same as for the formation of tetrasaccharides. The regioselective
glycosylation was accomplished using 3 eq of donor D5 which was reacted with tetrasaccharide
acceptor 10 to give hexasaccharide 11, isolated in 65% yield (Scheme 5). The subsequent O-
desilylation step was carried out by treatment with a 1 M solution of TBAF in THF followed by O-
acetylation using pyridine and acetic anhydride; compound 12 was isolated in 79% yield over two
steps. All O-acetyl groups were then removed using 1 M NaOMe in MeOH and compound 13 was
obtained in 82% yield. Hydrogenolysis employing Palladium on carbon (loading 10 – 20%) as a
catalyst furnished the hexasaccharide target compound 14 in 73% yield.
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Scheme 5. Formation of hexasaccharide 14. (a) i. 1 M TBAF, THF, 5 h, ii. Ac2O, DMAP, Pyr, 16 h,
92% over two steps; (b) DDQ, DCM/MeOH 4:1, 8 h, 85%; (c) TBDMSOTf, 2,6-lutidine, DCM,
−20 ºC → rt, 16 h, 95% ; (d) NIS, TfOH, DTBMP, DCM/Tol 1:1, −60 ºC → −30 ºC, 2 h, 65%; (e) i.
1 M TBAF, THF, 5 h, ii. Ac2O, DMAP, Pyr, 16 h, 79% over two steps; (f) 1 M NaOMe, MeOH, 16
h, 82%; (g) 10 – 20% Pd/C, H2 10 atm, MeOH, 16 h, 73%.
The structure of the synthesized hexasaccharide 14 was corroborated by high resolution mass
spectrometry data in which the pseudomolecular ion from an ESI-MS spectrum showed [M + Na]+
m/z 1027.3326, in excellent agreement with that calculated for C37H64O31Na, viz., m/z 1027.3329.
The 1H and 13C NMR resonances of 14 were fully assigned using 1D and 2D experiments (Table 3);
3
1
all transglycosidic JCH-based correlations anticipated in the H,13C-HMBC NMR spectrum were
1
observed, thereby leading further credence to the synthesized hexasaccharide structure. H NMR
n
1
chemical shifts and JHH coupling constants were refined by simulation of the H NMR spectrum
using an iterative total line-shape analysis,32 the result of which was in excellent agreement with the
experimental 1H NMR spectrum (Figure 3). The structures of the repeating units of the CPS from S.
pneumoniae type 37 and that the EPS from P. freudenreichii ssp. shermanii JS are the same24,25 and
for the latter 1H and 13C NMR chemical shift assignments have been reported. A comparison of the
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chemical shifts of the central disaccharide entity (B and B') of the hexasaccharide (cf. Figure 1) and
those of the EPS shows full agreement (Figure 4), in contrast to a pentasaccharide22 in which the
terminal C' residue is missing and key chemical shifts deviate from those of the polysaccharide.
Thus, a hexasaccharide model containing three repeating units is required to obtain agreement of
NMR data; consequently, the local environment at B and B' residues should represent the 3D
structure of the polysaccharide.
10
11
12
13
14
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Table 3. 1H and 13C NMR chemical shifts (ppm), nJHH and 1JH1,C1 (Hz) of hexasaccharide 14 at 9 °C.
4
5
Residue
1
2
3
4
5
6
3.739a
3.944b
1H
4.838
3.461
3.531
3.369
3.471
6
7
β-D-Glcp-(1→
C'
8
9
7.95
6.82a 1.96b
−12.35d
3JHH
9.46
9.18
9.99
(164)c
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
13C
1H
104.42
5.113
74.49
3.636
76.54
3.772
70.64
3.460
77.13
3.494
61.90
3.741a
3.935b
→2)-β-D-Glcp-(1→3)
7.87
6.06a 2.14b
−12.31d
3JHH
9.19
9.09
10.14
(~165)c
C
13C
1H
100.29
5.053
82.32
3.379
77.01
3.549
70.34
3.392
76.82
3.492
61.40
3.742a
3.928b
β-D-Glcp-(1→
B'
7.94
6.18a 1.32b
−12.44d
3JHH
9.52
9.02
10.00
(165)c
13C
1H
102.60
5.147
74.86
3.895
76.28
4.056
70.73
3.552
77.21
3.486
61.75
3.751a
3.927b
7.88
→2,3)-β-D-Glcp-(1→3)
5.57a 2.28b
−12.51d
3JHH
9.01
9.12
9.96
(~165)c
B
13C
1H
100.23
4.775
79.81
3.342
82.36
3.522
68.65
3.426
76.23
3.446
61.43
3.757a
3.920b
7.96
β-D-Glcp-(1→
A'
4.80a 2.03b
−12.26d
3JHH
9.45
8.77
9.97
(163)c
13C
1H
103.81
5.015
74.15
4.068
76.65
4.089
70.31
3.585
76.71
3.689
61.40
3.782a
3.886b
→2,3)-α-D-Glcp-OMee
3.77
5.24a 2.22b
71.63
−12.38d
3JHH
9.62
9.10
10.14
68.31
(175)c
A
13C
99.87
80.19
78.22
61.28
a H6pro-R; b H6pro-S. c 1JH1,C1 at 21 °C; d 2JHH; e OMe: δH 3.402, δC 55.46.
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1
Figure 3. H NMR spectrum at 700 MHz of hexasaccharide 14 at 9 °C (a) and the corresponding
spectrum simulated by total-lineshape analysis using the PERCH NMR software (b). The HDO
resonance in the experimental spectrum (δH 4.93) was removed prior to the lineshape fitting
procedure. Anomeric proton resonances are annotated according to residue.
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1
13
Figure 4. Comparison of H and C NMR chemical shifts (top and bottom, respectively) between
residues B and B' from hexasaccharide 14 and EPS from P. freudenreichii ssp. shermanii JS,25
which has the same polysaccharide structure as that of the CPS from S. pneumoniae type 37.24
CONCLUSIONS
In conclusion, a number of conformationally armed glycan building blocks have been introduced
and employed in convergent syntheses of tri- and tetrasaccharides. For some of these glucosyl
donors, excellent yields were obtained in the formation of trisaccharides. Due to the crowded nature
of the substrates, the reactions studied represent challenging systems and suitable reactions were
investigated. Orthogonal protective groups were used allowing selective functionalization of the
donors. The developed methodology was subsequently successfully applied in the synthesis of a
hexasaccharide representing three repeating units of CPS and EPS with β-(1→2)-linked side-chains
and a β-(1→3)-linked backbone. Notably, an NMR chemical shift analysis showed that a
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hexasaccharide is the smallest structure for which the central disaccharide entity (B and B')
represents a model for the above bacterial polysaccharides.
5
6
7
8
A conformational and structure-reactivity analysis was performed with the six disaccharide donors
D1 – D6 as well as the glycosylation products P3b and P4. This analysis rendered high quality data
1
through NMR spin-simulations of very complex H NMR spectra and population-weighted
9
averages could be produced when a single conformer was not sufficient to describe the
conformational space. The explored conformations deviate somewhat from those presented in the
literature for similar compounds, which have been designated as single conformers.11,14,30 The
present study indicates that a more elaborate description is necessary to structurally assign these
species; we foresee that by the use of low-temperature NMR studies one should be able to unravel
conformational preferences and elucidate dynamic equilibria between different ring
conformations,40 indicated to take place e.g. in donor D4. A correlation affirming that the more
axial-rich donors had a higher reactivity was clearly found, i.e., D2, D4 and D6 vs. D1, D3 and D5.
Furthermore, we hypothesize that the axial-rich skew and boat conformations for the particularly
reactive donors are pre-organized close to the anticipated transition states in the glycosylation
reactions. The structures and the corresponding structure-reactivity information obtained in this
study of super-armed disaccharide donors, in conjunction with previously established glycosylation
methodology, should be possible to apply efficiently in future challenging oligo- or polysaccharide
syntheses.
10
11
12
13
14
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EXPERIMENTAL SECTION
General methods
Dry solvents, including toluene (Tol), dichloromethane (DCM), tetrahydrofurane (THF) and
acetonitrile (ACN) were obtained from a VAC solvent purifier (Hawthorne, CA, USA). Dry N,N-
dimethylformamide (DMF) was purchased from Acros Organics (New Jersey, USA) and used as
received. Pyridine (Pyr) was distilled over CaH2 and dried with molecular sieves (4 Å). Methanol
(MeOH) was dried over molecular sieve (4 Å). All reagents were used as received. A nitrogen gas
flow was used for reactions requiring inert atmosphere. Powdered molecular sieves (4 Å) were
activated by heating under high vacuum. Column chromatography was performed on a Biotage
Isolera flash chromatography system (Uppsala, Sweden) using KP-Sil or HP-Sil snap silica gel
cartridges and purification on t-C18 Sep-pak® cartridges. TLC was carried out on silica gel 60 F254
plates (20 × 20 cm, 0.2 mm thickness) and monitored with UV light 254 – 360 nm or by a staining
solution prepared from Ceric Ammonium Sulfate (2 g) in ethanol (40 mL) and 2 M sulfuric acid (40
mL).
NMR spectra for characterization of all isolated compounds were recorded at 25 °C, unless
1
otherwise stated, on spectrometers operating at H frequencies of 400, 500, 600 or 700 MHz. The
NMR chemical shifts (δ) are reported in ppm and referenced to TMS as an internal standard, δH =
0.0, or the residual solvent peaks for CDCl3, δH = 7.26, or MeOH-d4, δH = 3.31. 13C chemical shifts
were referenced to external 1,4-dioxane in D2O, δC = 67.40, or internally to the CDCl3 residual
solvent peak δC = 77.16 or to the MeOH-d4-residual solvent peak δC = 49.00. J coupling constants
1
are reported in Hertz (Hz). All new compounds synthesized were fully characterized using 1D H,
1D 1H-decoupled 13C, 2D 1H,1H-DQF-COSY, 2D 1H,13C-multiplicity-edited-HSQC and 2D 1H,13C-
1
1
1
HMBC NMR experiments. If required, 1D H,1H-TOCSY, 1D H,1H-NOESY, 2D H,1H-TOCSY,
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2D H,13C-H2BC, 2D C,1H-HETCOR or 2D H,13C-HSQC-TOCSY experiments were acquired.
High-resolution mass spectra were recorded in positive mode on spectrometers using electrospray
ionization (ESI) equipped with time-of-flight (TOF) analyzers. Samples of 1 mg·mL−1 were
prepared using a solution of 1:1 ACN/H2O containing 0.1% formic acid.
6
7
8
9
Abbreviations for NMR resonances: br (broad), s (singlet), d (doublet), t (triplet), dd (doublet of
doublet), q (quadruplet), dt (doublet of triplet), dq (doublet of quadruplet), ddd (doublet of doublet
of doublet), m (multiplet). Of the two protons constituting the hydroxylmethyl group, the one
resonating at lower chemical shift is denoted H-6a, and the one at higher chemical shift is denoted
H-6b.
10
11
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General procedure for O-deacylation
The saccharide (x g, y mmol) was dissolved in MeOH (0.1 mM) and NaOMe in MeOH was added
(2 eq per acyl group using a 1 M solution) and the mixture was stirred at room temperature
overnight. When TLC (DCM/MeOH 9:1) indicated completion of the reaction, the solution was
stirred for 30 min with Dowex® 50W X8-H+ resin until pH 6 was reached. The resin was filtered off
and washed with methanol after which solvents were evaporated to yield O-deacylated products.
General procedure for per-O-silylation
The saccharide (x g, y mmol) was dissolved in dry Pyr to a concentration of ~0.03 mM and DMAP
(0.2 eq) was added to the mixture whereafter 3 molar equivalents of TBDMSOTf per alcohol group
to be protected were added dropwise at 0 °C. The reaction mixture was heated to 80 °C and stirred
overnight after which it was left to cool and subsequently quenched by addition of MeOH. The
mixture was extracted with DCM and successively washed with 1 M HCl, saturated aqueous
NaHCO3 and brine. The solution was dried over anhydrous Na2SO4 prior to solvent evaporation
under vacuum. The resulting residue was purified by flash chromatography to afford the respective
O-silylated disaccharide donor as colorless syrup.
General procedure for glycosylation
Donor (50 mg, y mmol), acceptor (3 eq) and base DTBMP (1.5 eq) were dissolved in dry solvents
(DCM/Tol 1:1) at room temperature to a concentration of ~0.04 mM with respect to the donor and
stirred with 4 Å molecular sieves under N2 flow. The temperature was decreased to −60 °C and NIS
(1.2 eq), previously dissolved in dry DCM, was added. TfOH (1.5 µL, cat amount) was
subsequently added at the same temperature. The mixture was allowed to reach −30 °C and stirred
for 30 min, while monitored by mass spectrometry analysis. The reaction was then quenched by the
addition of NEt3 (50 µL). Once room temperature was attained, the mixture was filtered through a
Celite pad and subsequently diluted into DCM. The resulting organic phase was washed with a 10%
Na2S2O3 solution and then with brine. The acquired mixture was dried over Na2S2O4 after which
solvents were evaporated. The obtained residue was purified by flash column chromatography
(Pentane/EtOAc 3:1) to yield tri- or tetrasaccharides as colorless oils.
Ethyl
2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→2)-3-O-(2-naphthyl)methyl-1-thio-β-D-
glucopyranoside (2).
Disaccharide 123 (0.3 g, 0.38 mmol) was dissolved in AcOH/H2O, 8:2 (5 mL). The reaction mixture
was heated at 80 °C during 4 h, monitored by TLC (Rf = 0.6 Tol/EtOAc 1:4). At completion, the
mixture was left to reach room temperature and then the reaction was quenched by NEt3 (0.5 mL).
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The solvents were evaporated and co-evaporated with toluene. The product was purified by flash
chromatography to yield the desired product 2, 220 mg (82% yield) as a colorless oil. H NMR
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(CDCl3, 25 °C, 400 MHz): Residue B: δ 7.95-7.44 (m, 7H, H-Ar), 5.05 (d, Jgem −11.05, 1H,
NapCH2), 4.90 (d, Jgem −11.05, 1H, NapCH2), 4.47 (d, JH1,H2 9.64, 1H, H-1), 3.86 (dd, JH5,H6b 3.72,
Jgem −11.95, 1H, H-6b), 3.76 (dd, JH5,H6a 4.78, Jgem −11.95, 1H, H-6a), 3.74 (dd, JH1,H2 9.64, JH2,H3
8.66, 1H, H-2), 3.66 (dd, JH3,H4 9.06, JH4,H5 9.28, 1H, H-4), 3.57 (dd, JH2,H3 8.66, JH3,H4 9.06, 1H, H-
3), 3.32 (ddd, JH4,H5 9.28, JH5,H6a 4.78, JH5,H6b 3.72, 1H, H-5), 2.70 (2q, 2H, SCH2CH3), 1.27 (t, 3H,
SCH2CH3). Residue B': 5.24-5.10 (m, 4H, H-1, H-2, H-3, H-4), 4.23 (dd, JH5,H6b 4.98, Jgem −12.37,
1H, H-6b), 4.13 (dd, JH5,H6a 2.56, Jgem −12.37, 1H, H-6a), 3.63 (ddd, JH4,H5 9.31, JH5,H6a 2.56, JH5,H6b
4.98, 1H, H-5), 2.08, 2.07, 2.02, 2.01 (4 s, 12H, CH3). 13C NMR (CDCl3, 25 °C, 100 MHz):
Residue B: 135.5, 133.5, 133.2 (3 C-ipso), 128.9, 128.1, 127.8, 127.3, 126.5, 126.4, 126.1 (7 C-Ar),
86.7 (C-2), 83.7 (C-1), 79.2 (C-5), 77.2 (C-3), 75.9 (NapCH2), 71.2 (C-4), 62.4 (C-6), 24.1
(SCH2CH3), 14.7 (SCH2CH3). Residue B': 170.8, 170.4, 169.5, 169.3 (4 C=O), 99.9 (C-1), 73.3 (C-
2), 72.1 (C-4), 71.8 (C-5), 68.5 (C-3), 62.1 (C-6), 21.0, 20.9, 20.7, 20.6 (4 CH3). ESI-HRMS: [M +
Na]+ m/z calc for C33H42O14SNa 717.2193, found 717.2191.
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Ethyl β-D-glucopyranosyl-(1→2)-3-O-(2-naphthylmethyl)-1-thio-β-D-glucopyranoside (3).
General procedure for O-deacylation: 2 eq per O-acyl group to be cleaved using a 1 M solution of
NaOMe in MeOH were added. Starting from ethyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-
(1→2)-3-O-(2-naphthyl)methyl-1-thio-β-D-glucopyranoside 2 (320 mg) the procedure yielded
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compound 3 (TLC: Rf = 0.35 DCM/MeOH 9:1) in 91% yield (220 mg) as a colorless syrup. H
NMR (CD3OD, 25 °C, 400 MHz): Residue B: δ 7.99-7.42 (m, 7H, H-Ar), 5.12 (s, 2H, NapCH2),
4.52 (d, JH1,H2 9.46, 1H, H-1), 3.88 (dd, JH5,H6b 2.26, Jgem −12.16, 1H, H-6b), 3.79 (dd, JH1,H2 9.46,
JH2,H3 8.62, 1H, H-2), 3.72 (dd, JH2,H3 8.62, JH3,H4 9.10, 1H, H-3), 3.67 (dd, JH5,H6a 6.03, Jgem −12.16,
1H, H-6a), 3.54 (dd, JH3,H4 9.10, JH4,H5 9.82, 1H, H-4), 3.34 (ddd, JH4,H5 9.82, JH5,H6a 6.03, JH5,H6b
2.26, 1H, H-5), 2.77 (2q, 2H, SCH2CH3), 1.27 (t, 3H, SCH2CH3). Residue B': 4.86 (d, JH1,H2 7.65,
1H, H-1), 3.81 (dd, JH5,H6b 2.49, Jgem −11.77, 1H, H-6b), 3.63 (dd, JH5,H6a 5.89, Jgem −11.77, 1H, H-
6a), 3.29-3.28 (m, 3H, H-3, H-4, H-2), 3.11 (ddd, JH4,H5 8.75, JH5,H6a 5.89, JH5,H6b 2.49, 1H, H-5).
13C NMR (CD3OD, 25 °C, 100 MHz): Residue B: δ 137.6, 134.9, 134.5 (3 C-ipso), 129.0, 128.8,
128.6, 128.1, 127.8, 127.0, 126.8 (7 C-Ar), 88.1 (C-3), 84.6 (C-1), 82.2 (C-5), 77.4 (C-2), 76.3
(NapCH2), 72.3 (C-4), 62.8 (C-6), 24.4 (SCH2CH3), 14.9 (SCH2CH3). Residue B': 103.6 (C-1), 78.0
(C-3), 78.0 (C-5), 75.4 (C-2), 71.8 (C-4), 63.0 (C-6). ESI-HRMS: [M + Na]+ m/z calc for
C25H34O10SNa 549.1770, found 549.1772.
Ethyl
2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-3-O-(2-
naphthylmethyl)-4,6-di-O-tert-butyldimethylsilyl-1-thio-β-D-glucopyranoside (D1).
General procedure for per-O-silylation: to hexaol 3 (92 mg) were added slowly, DMAP (0.2 eq) and
TBDMSOTf (3 eq per alcohol group to be protected) at 0 °C under N2 atmosphere and the mixture
was heated to 80 °C overnight. The product was purified by chromatography (TLC: Rf = 0.5
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Pentane/DCM 1:1) to afford thioglycoside donor D1 in 85% yield (180 mg) as a colorless oil. H
NMR (CDCl3, 25 °C, 400 MHz): Residue B: δ 7.84-7.13 (m, 7H, H-Ar), 5.25 (d, Jgem −11.81, 1H,
NapCH2), 4.76 (d, Jgem −11.81, 1H, NapCH2), 4.69 (d, JH1,H2 9.19, 1H, H-1), 3.94 (dd, JH1,H2 9.19,
JH2,H3 7.99, 1H, H-2), 3.87 (dd, Jgem −11.14, 1H, H-6b), 3.80 (dd, JH2,H3 7.99, JH3,H4 8.24, 1H, H-3),
3.72 (dd, Jgem −11.14, 1H, H-6a), 3.66 (dd, JH3,H4 8.24, JH4,H5 9.29, 1H, H-4), 3.32 (ddd, JH4,H5 9.29,
1H, H-5), 2.70 (2q, 2H, SCH2CH3), 1.26 (t, 3H, SCH2CH3). Residue B': 5.25 (d, JH1,H2 5.95, 1H, H-
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1), 3.81 (ddd, JH4,H5 1.70, 1H, H-5), 3.80 (dd, Jgem −9.94, 1H, H-6b), 3.80 (dd, JH3,H4 3.47, JH4,H5
1.70, 1H, H-4), 3.72 (dd, Jgem −9.94, 1H, H-6a), 3.66 (dd, JH2,H3 0.58, JH3,H4 3.47, 1H, H-3), 3.62
(dd, JH1,H2 5.95, JH2,H3 0.58, 1H, H-2); 0.92, 0.90, 0.84, 0.83, 0.80, 0.78 (6 s, 54H, C(CH3)3), 0.11,
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0.10, 0.08, 0.06, 3 × 0.02, 0.01, 2 × −0.02, −0.03, −0.10 (12 s, 36H, SiCH3). C NMR (CDCl3, 25
°C, 100 MHz): Residue B: δ 137.7, 134.3, 133.7 (3 C-ipso), 128.8, 2 × 128.4, 126.4, 126.3, 126.2,
126.1 (7 C-Ar), 87.5 (C-3), 83.2 (C-1), 81.7 (C-5), 77.7 (C-2), 71.6 (C-4), 74.8 (NapCH2), 63.7 (C-
6), 25.1 (SCH2CH3), 15.7 (SCH2CH3). Residue B': 99.6 (C-1), 83.0 (C-5), 79.4 (C-3), 79.1 (C-2),
71.6 (C-4), 65.8 (C-6); 26.2, 3 × 26.1, 2 × 25.9 (6 C(CH3)3), 2 × 18.6, 2 × 18.2, 2 × 17.9 (6
C(CH3)3), −2.9, −3.0, −3.1, −3.5, 3 × −3.6, −3.9, −4.0, −4.1, −4.2, −4.4 (12 SiCH3). ESI-HRMS: [M
+ Na]+ m/z calc for C61H118O10SSi6Na 1233.6959, found 1233.6956.
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Ethyl
β-D-glucopyranosyl-(1→2)-3-O-(2-naphthylmethyl)-6-O-benzyl-1-thio-β-D-
glucopyranoside (5).
General procedure for O-deacylation: 2 eq per O-acyl group to be cleaved using a 1 M solution of
NaOMe in MeOH were added. Starting from ethyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-
(1→2)-3-O-(2-naphthylmethyl)-6-O-benzyl-1-thio-β-D-glucopyranoside23 4 (250 mg) yielded
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compound 5 (Rf = 0.45 DCM/MeOH 9:1) in 92% yield (181 mg) as a colorless syrup. H NMR
(CD3OD, 25 °C, 400 MHz): Residue B: δ 7.99-7.23 (m, 12H, H-Ar), 5.12 (s, 2H, NapCH2), 4.59 (s,
2H, PhCH2), 4.54 (d, JH1,H2 9.46, 1H, H-1), 3.84 (dd, JH5,H6b 1.95, Jgem −11.10, 1H, H-6b), 3.80 (dd,
JH1,H2 9.46, JH2,H3 8.78, 1H, H-2), 3.73 (dd, JH2,H3 8.78, JH3,H4 8.60, 1H, H-3), 3.67 (dd, JH5,H6a 5.83,
Jgem −11.10, 1H, H-6a), 3.59 (dd, JH3,H4 8.60, JH4,H5 9.94, 1H, H-4), 3.50 (ddd, JH4,H5 9.94, JH5,H6a
5.83, JH5,H6b 1.95, 1H, H-5), 2.74 (2q, 2H, SCH2CH3), 1.27 (t, 3H, SCH2CH3). Residue B': 4.85 (d,
1H, H-1), 3.81 (dd, JH5,H6b 2.56, Jgem −11.82, 1H, H-6b), 3.64 (dd, JH5,H6a 5.81, Jgem −11.82, 1H, H-
6a), 3.32-3.28 (m, 3H, H-3, H-4, H-2), 3.12 (ddd, JH4,H5 8.86, JH5,H6a 5.81, JH5,H6b 2.56, 1H, H-5).
13C NMR (CD3OD, 25 °C, 100 MHz): Residue B: δ 139.7, 137.6, 134.8, 134.4 (4 C-ipso), 2 ×
129.3, 129.0, 3 × 128.8, 2 × 128.6, 128.1, 127.7, 127.0, 126.8 (12 C-Ar), 88.0 (C-3), 84.6 (C-1),
81.0 (C-5), 77.5 (C-2), 76.3 (NapCH2), 74.4 (PhCH2), 72.4 (C-4), 70.8 (C-6), 24.6 (SCH2CH3), 15.2
(SCH2CH3). Residue B': 103.5 (C-1), 78.0 (C-3), 78.0 (C-5), 75.4 (C-2), 71.8 (C-4), 63.0 (C-6).
ESI-HRMS: [M + Na]+ m/z calc for C32H40O10SNa 639.2240, found 639.2238.
Ethyl
2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-3-O-(2-
naphthylmethyl)-4-O-tert-butyldimethylsilyl-6-O-benzyl-1-thio-β-D-glucopyranoside (D3).
General procedure for per-O-silylation: to pentaol 5 (85 mg) were added slowly, DMAP (0.2 eq)
and TBDMSOTf (3 eq per alcool group to be protected) at 0 °C under N2 atmosphere and the
mixture was heated to 80 °C overnight. The product was purified by chromatography (TLC: Rf =
0.8 Pentane/DCM 2:1) to afford thioglycoside donor D3 in 86% yield (141 mg) as a colorless oil.
1H NMR (CDCl3, 25 °C, 400 MHz): Residue B: δ 7.82-7.24 (m, 12H, H-Ar), 5.23 (d, Jgem −11.44,
1H, NapCH2), 4.76 (d, Jgem −11.44, 1H, NapCH2), 4.76 (d, JH1,H2 8.92, 1H, H-1), 4.65 (d, Jgem
−12.20, 1H, PhCH2), 4.52 (d, Jgem −12.20, 1H, PhCH2), 4.01 (dd, JH1,H2 8.92, JH2,H3 7.73, 1H, H-2),
3.82 (dd, JH2,H3 7.73, JH3,H4 8.18, 1H, H-3), 3.77 (dd, Jgem −10.47, 1H, H-6b), 3.69 (dd, JH3,H4 8.18,
JH4,H5 9.46, 1H, H-4), 3.57 (dd, Jgem −10.47, 1H, H-6a), 3.57 (ddd, JH4,H5 9.46, 1H, H-5), 2.76 (q,
2H, SCH2CH3), 1.30 (t, 3H, SCH2CH3). Residue B': 5.24 (d, JH1,H2 6.00, 1H, H-1), 3.81 (dd,
JH3,H4 3.40, JH4,H5 1.55, 1H, H-4), 3.81 (ddd, JH4,H5 1.55, 1H, H-5), 3.81 (dd, Jgem −9.96, 1H, H-6b),
3.73 (dd, Jgem −9.96, 1H, H-6a), 3.67 (dd, JH2,H3 0.54, JH3,H4 3.40, 1H, H-3), 3.63 (dd, JH1,H2 6.00,
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JH2,H3 0.54, 1H, H-2); 0.93, 0.85, 0.81, 0.80, 0.78 (5 s, 45H, C(CH3)3), 0.12, 0.11, 0.05, 0.03, 0.01,
−0.005, −0.02, −0.03, −0.05, −0.12 (10 s, 30H, SiCH3). 13C NMR (CDCl3, 25 °C, 100 MHz):
Residue B: δ 138.7, 136.8, 133.5, 132.9 (4 C-ipso), 2 × 128.4, 128.1, 127.8, 3 × 127.7, 127.5, 125.8,
125.6, 125.5, 125.4 (12 C-Ar), 86.4 (C-3), 82.8 (C-1), 79.7 (C-5), 76.7 (C-2), 73.9 (NapCH2), 73.4
(PhCH2), 71.4 (C-4), 70.1 (C-6), 24.6 (SCH2CH3), 15.0 (SCH2CH3). Residue B': 98.8 (C-1), 82.3
(C-5), 78.6 (C-3), 78.4 (C-2), 70.7 (C-4), 64.9 (C-6); 26.2, 26.1, 26.0, 2 × 25.9 (5 C(CH3)3), 18.6, 2
× 18.2, 2 × 17.9 (5 C(CH3)3), −3.7, −3.8, −3.9, −4.4, 2 × −4.6, 2 × −4.7, −4.9, −5.1 (10 SiCH3). ESI-
HRMS: [M + Na]+ m/z calc for C62H110O10SSi5Na 1209.6564, found 1209.6567.
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Ethyl 6-O-benzyl-β-D-glucopyranosyl-(1→2)-3-O-(2-naphthylmethyl)-6-O-benzyl-1-thio-β-D-
glucopyranoside (7).
General procedure for O-deacylation: 2 eq per acyl group to be cleaved using a 1 M solution of
NaOMe in MeOH were added. Starting from ethyl 2,3,4-tri-O-benzoyl-6-O-benzyl-β-D-
glucopyranosyl-(1→2)-3-O-(2-naphthylmethyl)-6-O-benzyl-1-thio-β-D-glucopyranoside 623 (210
mg) yielded compound 7 (Rf = 0.55 DCM/MeOH 9:1) in 92% yield (134 mg) as a colorless syrup.
1H NMR (CD3OD, 25 °C, 400 MHz): δ 8.00-7.17 (m, 17H, H-Ar); Residue B: 5.12 (s, 2H,
NapCH2), 4.59 (s, 2H, PhCH2), 4.53 (d, JH1,H2 9.45, 1H, H-1), 3.84 (dd, JH5,H6b 1.93, Jgem −11.11,
1H, H-6b), 3.80 (dd, JH1,H2 9.45, JH2,H3 8.74, 1H, H-2), 3.74 (dd, JH2,H3 8.74, JH3,H4 8.57, 1H, H-3),
3.67 (dd, JH5,H6a 5.93, Jgem −11.11, 1H, H-6a), 3.58 (dd, JH3,H4 8.57, JH4,H5 9.88, 1H, H-4), 3.49 (ddd,
JH4,H5 9.88, JH5,H6a 5.93, JH5,H6b 1.93, 1H, H-5), 2.71 (2q, 2H, SCH2CH3), 1.22 (t, 3H, SCH2CH3).
Residue B': 4.88 (d, 1H, H-1), 4.60 (2d, Jgem −11.97, 2H, PhCH2), 3.81 (dd, JH5,H6b 2.00, Jgem
−11.29, 1H, H-6b), 3.64 (dd, JH5,H6a 5.93, Jgem −11.29, 1H, H-6a), 3.36-3.29 (m, 3H, H-3, H-4, H-2),
3.23 (ddd, JH4,H5 9.31, JH5,H6a 5.93, JH5,H6b 2.00, 1H, H-5). 13C NMR (CD3OD, 25 °C, 100 MHz): δ
139.9, 139.7, 137.6, 134.8, 134.4 (5 C-ipso), 130.4, 129.6, 4 × 129.3, 129.0, 4 × 128.8, 2 × 128.6,
128.4, 128.1, 127.7, 127.0 (17 C-Ar); Residue B: 88.2 (C-3), 84.8 (C-1), 81.0 (C-5), 77.2 (C-2),
76.3 (NapCH2), 74.4 (PhCH2), 72.4 (C-4), 70.8 (C-6), 24.5 (SCH2CH3), 15.3 (SCH2CH3). Residue
B': 103.5 (C-1), 78.1 (C-3), 77.4 (C-5), 75.3 (C-2), 71.8 (C-4), 70.8 (C-6). ESI-HRMS: [M + Na]+
m/z calc for C39H46O10SNa 729.2709, found 729.2706.
Ethyl
2,3,4-tri-O-tert-butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→2)-3-O-(2-
naphthylmethyl)-4-O-tert-butyldimethylsilyl-6-O-benzyl-1-thio-β-D-glucopyranoside (D5).
General procedure for per-O-silylation: to tetraol 7 (105 mg) were added slowly, DMAP (0.2 eq)
and TBDMSOTf (3 eq per alcool group to be protected) at 0 °C under N2 atmosphere and the
mixture was heated to 80 °C overnight. The product was purified by chromatography (TLC: Rf =
0.75 Pentane/DCM 2:1) to afford thioglycoside donor D5 in 88% yield (152 mg) as a colorless oil.
1H NMR (CDCl3, 25 °C, 400 MHz): δ 7.82-7.24 (m, 17H, H-Ar); Residue B: 5.22 (d, Jgem −11.89,
1H, NapCH2), 4.77 (d, Jgem −11.89, 1H, NapCH2), 4.70 (d, JH1,H2 9.02, 1H, H-1), 4.64 (d, Jgem
−12.16, 1H, PhCH2), 4.53 (d, Jgem −12.16, 1H, PhCH2), 4.00 (dd, JH1,H2 9.02, JH2,H3 7.98, 1H, H-2),
3.78 (dd, JH2,H3 7.98, JH3,H4 8.23, 1H, H-3), 3.76 (dd, Jgem −10.60, 1H, H-6b), 3.66 (dd, JH3,H4 8.23,
JH4,H5 9.50, 1H, H-4), 3.57 (dd, Jgem −10.60, 1H, H-6a), 3.53 (ddd, JH4,H5 9.50, 1H, H-5), 2.75 (2q,
2H, SCH2CH3), 1.30 (t, 3H, SCH2CH3). Residue B': 5.24 (d, JH1,H2 5.43, 1H, H-1), 4.65 (d, Jgem
−12.08, 1H, PhCH2), 4.55 (d, Jgem −12.08, 1H, PhCH2), 3.99 (ddd, JH4,H5 2.95, 1H, H-5), 3.85 (dd,
JH3,H4 3.41, JH4,H5 2.95, 1H, H-4), 3.70 (dd, Jgem −9.67, 1H, H-6b), 3.65 (dd, JH1,H2 5.43, JH2,H3 0.73,
1H, H-2), 3.65 (dd, JH2,H3 0.73, JH3,H4 3.41, 1H, H-3), 3.64 (dd, Jgem −9.67, 1H, H-6a); 0.81, 0.79,
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0.77, 0.75 (4 s, 36H, C(CH3)3), 0.07, −0.01, −0.04, 2 × −0.05, −0.06, −0.07, −0.14 (8 s, 24H,
SiCH3). 13C NMR (CDCl3, 25 °C, 100 MHz): δ 2 × 138.7, 136.7, 133.5, 132.9 (5 C-ipso), 4 ×
128.4, 128.1, 3 × 127.8, 3 × 127.7, 2 × 127.6, 125.8, 125.5, 2 × 125.4 (17 C-Ar); Residue B: 86.8
(C-3), 83.1 (C-1), 80.0 (C-5), 76.6 (C-2), 74.0 (NapCH2), 73.5 (PhCH2), 71.4 (C-4), 70.1 (C-6),
24.7 (SCH2CH3), 15.1 (SCH2CH3). Residue B': 99.2 (C-1), 79.4 (C-5), 78.6 (C-3), 77.6 (C-2), 73.4
(PhCH2), 71.9 (C-6), 71.5 (C-4); 26.1, 26.0, 2 × 25.9 (4 C(CH3)3), 2 × 18.1, 2 × 17.9 (4 C(CH3)3),
−3.7, 2 × −4.0, −4.3, −4.4, −4.6, 2 × −4.7 (8 SiCH3). ESI-HRMS: [M + Na]+ m/z calc for
C63H102O10SSi4Na 1185.6168, found 1185.6164.
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Methyl
2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→2)-4,6-O-benzylidene-α-D-
glucopyranoside (A3).
A solution of 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl trichloroacetimidate41 (245 mg, 2 eq) and
methyl 3-O-p-methoxybenzyl-4,6-O-benzylidene-α-D-glucopyranoside42 (100 mg) in dry CH2Cl2 (4
mL) was stirred for 20 min in the presence of 4 Å molecular sieves. TMSOTf (0.08 eq) was added
at −40 °C. The reaction was monitored by TLC (pentane/EtOAc 1:2) and allowed to warm up to
room temperature. NEt3 was added to quench the reaction and the mixture was filtered through a
pad of Celite. It was then diluted with CH2Cl2 and washed successively with solutions of saturated
NaHCO3 and brine and dried over Na2SO4. The solvent was removed in vaccuo, and the residue
obtained was purified by flash chromatography (TLC: Rf = 0.79 Pentane/EtOAc 1:2) to yield the
precursor
disaccharide
methyl
2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→2)-3-O-p-
methoxybenzyl-4,6-O-benzylidene-α-D-glucopyranoside (A3p) in 92% yield (168 mg) as a white
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powder. H NMR (CDCl3, 25 °C, 400 MHz): Residue A: δ 7.48-6.83 (m, 9H, H-Ar), 5.54 (s, 1H,
CHPh), 4.83 (d, JH1,H2 3.66, 1H, H-1), 4.73 (d, Jgem −10.58, 1H, CH3OArCH2), 4.64 (d, Jgem −10.58,
1H, CH3OArCH2), 4.28 (dd, JH5,H6a 10.26, JH5,H6b 4.68, Jgem −10.01, 1H, H-6b), 3.98 (dd, JH2,H3 9.43,
JH3,H4 9.29, 1H, H-3), 3.84 (ddd, JH4,H5 9.36, JH5,H6a 5.86, JH5,H6b 4.68, 1H, H-5), 3.79 (s, 3H,
CH3OArCH2), 3.73 (dd, JH5,H6a 10.26, Jgem −10.01, 1H, H-6a), 3.71 (dd, JH1,H2 3.66, JH2,H3 9.43, 1H,
H-2), 3.66 (dd, JH3,H4 9.29, JH4,H5 9.36, 1H, H-4), 3.42 (s, 3H, OMe). A': 5.20 (dd, JH2,H3 9.36, JH3,H4
9.30, 1H, H-3), 5.12 (dd, JH1,H2 7.80, JH2,H3 9.36, 1H, H-2), 5.10 (dd, JH3,H4 9.30, JH4,H5 9.92, 1H, H-
4), 4.87 (d, JH1,H2 7.80, 1H, H-1), 4.18 (m, 2H, H-6b, H-6a), 3.67 (ddd, 1H, H-5), 2.08, 2.03, 2.00,
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1.91 (4s, 12H, CH3). C NMR (CDCl3, 25 °C, 100 MHz): Residue A: δ 158.5 (C-para), 137.5,
130.6 (2 C-ipso), 2 × 129.7, 129.1, 2 × 128.4, 2 × 126.2, (7 C-Ar), 114.0 (2 C-Ar), 101.5 (CHPh),
100.2 (C-1), 82.4 (C-4), 80.2 (C-2), 77.3 (C-3), 75.0 (CH3OArCH2), 69.3 (C-6), 62.4 (C-5), 55.6
(OMe), 55.4 (CH3OArCH2). A': 170.7, 170.5, 169.5, 169.3 (4 C=O), 101.8 (C-1), 73.3 (C-3), 72.1
(C-5), 71.7 (C-2), 68.6 (C-4), 62.2 (C-6), 20.9, 2 × 20.8, 20.7 (4 CH3). ESI-HRMS: [M + Na]+ m/z
calc for C36H44O16Na 755.2527, found 755.2525.
To a solution of A3p (163 mg) in CH2Cl2 (3 ml) was added water (0.3 ml) and 2,3-dichloro-5,6-
dicyano-1,4-benzoquinone (DDQ, 76 mg, 1.5 eq).43 The reaction mixture was stirred at room
temperature for 2 h. The mixture was diluted with CH2Cl2 and washed twice with a solution of
saturated NaHCO3 and then dried over Na2SO4. The solvent was evaporated, and the oil obtained
was purified by flash chromatography (TLC: Rf = 0.65 Pentane/EtOAc 1:2) to yield the acceptor
methyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→2)-4,6-O-benzylidene-α-D-glucopyranoside
1
A3 in 88% yield (120 mg) as a white powder. H NMR (CDCl3, 25 °C, 400 MHz): Residue A: δ
7.51-7.33 (m, 5H, H-Ar), 5.53 (s, 1H, CHPh), 4.82 (d, JH1,H2 3.69, 1H, H-1), 4.29 (dd, JH5,H6b 4.70,
Jgem −10.10, 1H, H-6b), 4.13 (ddd, JH2,H3 9.42, JH3,H4 9.20, JH3,OH3 1.67, 1H, H-3), 3.85 (ddd, JH4,H5
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9.40, JH5,H6a 10.35, JH5,H6b 4.70, 1H, H-5), 3.73 (dd, JH5,H6a 10.35, Jgem −10.10, 1H, H-6a), 3.61 (dd,
JH1,H2 3.69, JH2,H3 9.42, 1H, H-2), 3.51 (dd, JH3,H4 9.20, JH4,H5 9.40, 1H, H-4), 3.42 (s, 3H, OMe),
2.83 (d, JH3,OH3 1.67, 1H, OH-3). A': 5.23 (dd, JH2,H3 9.60, JH3,H4 9.48, 1H, H-3), 5.05 (dd,
JH1,H2 7.98, JH2,H3 9.60, 1H, H-2), 5.05 (dd, JH3,H4 9.48, JH4,H5 10.05, 1H, H-4), 4.84 (d, JH1,H2 7.98,
1H, H-1), 4.21 (dd, JH5,H6b 2.68, Jgem −12.27, 1H, H-6b), 4.15 (dd, JH5,H6a 5.64, Jgem −12.27, 1H, H-
6a), 3.72 (ddd, JH4,H5 10.05, JH5,H6a 5.64, JH5,H6b 2.68, 1H, H-5), 2.08, 2 × 2.03, 2.00 (4 s, 12H, CH3).
13C NMR (CDCl3, 25 °C, 100 MHz): Residue A: δ 137.2 (C-ipso), 129.3, 2 × 128.4, 2 × 126.4, (5
C-Ar), 102.0 (CHPh), 100.0 (C-1), 81.8 (C-2), 81.3 (C-4), 69.2 (C-3), 69.1 (C-6), 62.2 (C-5), 55.7
(OMe). A': 170.7, 170.3, 169.6, 169.5 (4 C=O), 101.7 (C-1), 72.7 (C-3), 72.0 (C-5), 71.6 (C-2),
68.6 (C-4), 62.2 (C-6), 2 × 20.8, 2 × 20.7 (4 CH3). ESI-HRMS: [M + Na]+ m/z calc for C28H36O15Na
635.1952, found 635.1948.
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2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-1,6-anhydro-3-O-(2-
naphthylmethyl)-4-O-tert-butyldimethylsilyl-β-D-glucopyranose (B1).
General procedure for Glycosylation: from Donor D1 (50 mg) and acceptor A1 (40 mg). The
obtained mixture was purified by flash column chromatography (Rf = 0.85 Pentane/DCM 2:1) to
yield disaccharide B1 in 48% yield (21 mg) as a colorless oil. 1H NMR (CDCl3, 25 °C, 400 MHz):
Residue B: δ 7.85-7.41 (m, 7H, H-Ar), 5.72 (d, JH1,H2 1.92, 1H, H-1), 4.83 (d, Jgem −11.71, 1H,
NapCH2), 4.68 (d, Jgem −11.71, 1H, NapCH2), 4.39 (ddd, JH5,H6b 1.00, 1H, H-5), 3.95 (dd, JH5,H6b
1.00, Jgem −7.11, 1H, H-6b), 3.77 (dd, 1H, H-3), 3.73 (dd, 1H, H-4), 3.67 (dd, Jgem −7.11, 1H, H-
6a), 3.64 (dd, JH1,H2 1.92, 1H, H-2). Residue B': 4.89 (d, JH1,H2 5.34, 1H, H-1), 3.92 (dd, 1H, H-4),
3.76 (dd, 1H, H-3), 3.76 (dd, 1H, H-6b), 3.73 (dd, 1H, H-6a), 3.66 (dd, 1H, H-2), 3.63 (ddd, 1H, H-
5); 2 × 0.89, 0.88, 0.87, 0.82 (5 s, 45H, C(CH3)3), 0.10, 0.08, 0.07, 2 × 0.06, 3 × 0.05, 0.04, −0.08
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(10 s, 30H, SiCH3). C NMR (CDCl3, 25 °C, 100 MHz): Residue B: δ 135.7, 133.4, 133.1 (3 C-
ipso), 128.3, 128.0, 127.8, 2 × 126.2, 125.9, 125.6 (7 C-Ar), 102.3 (C-1), 81.2 (C-3), 77.6 (C-2),
77.4 (C-5), 72.7 (C-4), 72.3 (NapCH2), 65.5 (C-6). Residue B': 103.2 (C-1), 81.2 (C-5), 78.8 (C-3),
77.2 (C-2), 70.1 (C-4), 63.9 (C-6); 3 × 26.1, 2 × 26.0 (5 C(CH3)3), 2 × 18.5, 18.1, 2 × 18.0 (5
C(CH3)3), −3.6, −4.0, 2 × −4.3, 2 × −4.5, 2 × −4.6, 2 × −5.2 (10 SiCH3). ESI-HRMS: [M + Na]+ m/z
calc for C53H98O10Si5Na 1057.5904, found 1057.5908.
2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-1,6-anhydro-3,4-di-O-tert-
butyldimethylsilyl-β-D-glucopyranose (B2).
General procedure for Glycosylation: from Donor D2 (50 mg) and acceptor A1 (41 mg). The
obtained mixture was purified by flash column chromatography (Rf = 0.55 Pentane/DCM 2:1) to
yield disaccharide B2 in 65% yield (28 mg) as a colorless oil. 1H NMR (CDCl3, 25 °C, 400 MHz):
Residue B: δ 5.72 (d, JH1,H2 1.55, 1H, H-1), 4.38 (ddd, JH4,H5 1.47, JH5,H6a 6.10, JH5,H6b 1.20, 1H, H-
5), 4.04 (dd, JH5,H6b1.20, Jgem −6.80, 1H, H-6b), 3.81 (dd, JH2,H3 0.66, JH3,H4 1.05, 1H, H-3), 3.66 (dd,
JH5,H6a 6.10, Jgem −6.80, 1H, H-6a), 3.52 (dd, JH3,H4 1.05, JH4,H5 1.47, 1H, H-4), 3.33 (dd, JH1,H2 1.55,
JH2,H3 0.66, 1H, H-2). Residue B': 4.82 (d, JH1,H2 5.88, 1H, H-1), 3.92 (dd, JH3,H4 2.91, JH4,H5 1.47,
1H, H-4), 3.76 (dd, JH2,H3 0.71, JH3,H4 2.91, 1H, H-3), 3.75 (ddd, JH4,H5 1.47, JH5,H6a 5.31, JH5,H6b
8.59, 1H, H-5), 3.71 (dd, JH5,H6b 8.59, Jgem −9.93, 1H, H-6b), 3.67 (dd, JH1,H2 5.88, JH2,H3 0.71, 1H,
H-2), 3.66 (dd, JH5,H6a 5.31, Jgem −9.93, 1H, H-6a); 0.93, 2 × 0.89, 3 × 0.88, (6s, 54H, C(CH3)3),
0.12, 0.11, 3 × 0.10, 0.09, 2 × 0.08, 2 × 0.07, 2 × 0.04 (12s, 36H, SiCH3). 13C NMR (CDCl3, 25 °C,
100 MHz): Residue B: δ 102.2 (C-1), 79.7 (C-2), 76.4 (C-5), 73.9 (C-3), 73.7 (C-4), 64.5 (C-6).
Residue B': 104.4 (C-1), 81.6 (C-5), 79.4 (C-3), 77.0 (C-2), 70.0 (C-4), 63.9 (C-6); 26.4, 2 × 26.1, 2
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× 26.0, 25.8 (6 C(CH3)3), 18.7, 18.5, 18.1, 18.0, 2 × 17.9 (6 C(CH3)3), −2.9, −3.9, −4.0, −4.3, 2 ×
−4.4, −4.5, 2 × −4.6, −4.8, 2 × −5.1 (12 SiCH3). ESI-HRMS: [M + Na]+ m/z calc for
C48H104O10Si6Na 1031.6143, found 1031.6146.
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Methyl
2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-3-O-(2-
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naphthylmethyl)-4-O-tert-butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→3)-4,6-O-
benzylidene-2-O-acetyl-α-D-glucopyranoside (P3a).
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General procedure for Glycosylation: from Donor D3 (50 mg) and acceptor A1 (41 mg). The
obtained product was purified by flash column chromatography (Rf = 0.55 Pentane/EtOAc 9:1) to
yield trisaccharide P3a in 50% yield (31 mg) as a colorless oil. 1H NMR (CDCl3, 25 °C, 400 MHz):
δ 7.85-7.21 (m, 17H, H-Ar); Residue B': 5.02 (d, JH1,H2 6.29, 1H, H-1), 3.96 (dd, 1H, H-3), 3.78
(dd, 1H, H-4), 3.76 (ddd, 1H, H-5), 3.71 (dd, 1H, H-6b), 3.67 (dd, 1H, H-6a), 3.62 (dd, JH1,H2 6.29,
1H, H-2); Residue B: 5.24 (d, JH1,H2 2.75, 1H, H-1), 4.86 (d, Jgem −11.71, 1H, NapCH2), 4.55 (d,
Jgem −12.40, 1H, PhCH2), 4.54 (d, Jgem −11.71, 1H, NapCH2), 4.31 (d, Jgem −12.40, 1H, PhCH2),
4.09 (dd, JH1,H2 2.75, 1H, H-2), 4.02 (dd, 1H, H-4), 3.81 (ddd, 1H, H-5), 3.48 (dd, 1H, H-3), 3.48
(dd, 1H, H-6b), 3.41 (dd, 1H, H-6a); Residue A: 5.43 (s, 1H, CHPh), 4.88 (d, JH1,H2 3.71, 1H, H-1),
4.84 (dd, JH1,H2 3.71, JH2,H3 9.39, 1H, H-2), 4.40 (dd, JH2,H3 9.39, JH3,H4 9.32, 1H, H-3), 4.21 (dd,
JH5,H6b 4.71, Jgem −10.20, 1H, H-6b), 3.84 (ddd, JH5,H6b 4.71, 1H, H-5), 3.62 (dd, Jgem −10.20, 1H, H-
6a), 3.61 (dd, JH3,H4 9.32, 1H, H-4), 3.37 (s, 3H, OMe), 1.99 (s, 1H, CH3); 0.88, 0.87, 0.86, 0.83,
0.66 (5 s, 45H, C(CH3)3), 2 × 0.12, 0.09, 0.08, 0.05, 0.01, −0.01, −0.02, −0.15, −0.32 (10 s, 30H,
SiCH3). 13C NMR (CDCl3, 25 °C, 100 MHz): δ 139.2, 137.6, 136.0, 133.4, 133.0 (5 C-ipso), 129.2,
2 × 128.4, 2 × 128.3, 128.1, 127.9, 127.8, 2 × 127.6, 127.3, 2 × 126.8, 2 × 126.0, 2 × 125.7 (17 C-
Ar); Residue B': 98.8 (C-1), 82.6 (C-5), 78.9 (C-4), 77.8 (C-2), 69.8 (C-3), 64.0 (C-6); Residue B:
99.7 (C-1), 84.6 (C-3), 74.7 (C-2), 74.3 (C-5), 72.9 (PhCH2), 71.0 (NapCH2), 71.3 (C-6), 70.0 (C-
4); Residue A: 170.4 (C=O), 102.3 (CHPh), 97.8 (C-1), 80.3 (C-4), 74.6 (C-2), 72.8 (C-3), 69.1 (C-
6), 62.7 (C-5), 55.4 (OMe), 21.1 (CH3).; 26.2, 26.1, 26.0, 2 × 25.9 (5 C(CH3)3), 18.4, 18.2, 18.1, 2 ×
18.0, (5 C(CH3)3), −3.7, −3.8, −4.1, −4.3, −4.4, −4.6, −4.7, 3 × −5.2 (10 SiCH3). ESI-HRMS: [M +
Na]+ m/z calc for C76H124O17Si5Na 1471.7583, found 1471.7582.
Methyl
2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-3-O-(2-
naphthylmethyl)-4-O-tert-butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→3)-4,6-O-
benzylidene-2-O-benzyl-α-D-glucopyranoside (P3b).
General procedure for Glycosylation: from Donor D3 (50 mg) and acceptor A2 (47 mg). The
obtained product was purified by flash column chromatography (Rf = 0.75 Pentane/EtOAc 9:1) to
1
yield trisaccharide P3b in 70% yield (45 mg) as a white powder. H NMR (CDCl3, 25 °C, 400
MHz): δ 7.84-7.13 (m, 22H, H-Ar); Residue B': 5.13 (d, JH1,H2 6.60, 1H, H-1), 4.02 (dd, 1H, H-3),
3.84 (ddd, 1H, H-5), 3.80 (dd, 1H, H-4), 3.71 (dd, 1H, H-6b), 3.67 (dd, JH1,H2 6.60, 1H, H-2), 3.60
(dd, 1H, H-6a); Residue B: 5.42 (d, JH1,H2 2.95, 1H, H-1), 4.91 (d, Jgem −11.66, 1H, NapCH2), 4.66
(d, Jgem −11.66, 1H, NapCH2), 4.58 (d, Jgem −12.56, 1H, PhCH2), 4.35 (d, Jgem −12.56, 1H, PhCH2),
4.24 (dd, JH1,H2 2.95, 1H, H-2), 4.22 (dd, 1H, H-4), 3.84 (ddd, 1H, H-5), 3.56 (dd, 1H, H-3), 3.49
(dd, 1H, H-6b), 3.48 (dd, 1H, H-6a); Residue A: 5.39 (s, 1H, CHPh), 4.61 (d, Jgem −12.49, 1H,
PhCH2), 4.48 (d, Jgem −12.49, 1H, PhCH2), 4.34 (d, JH1,H2 3.66, 1H, H-1), 4.28 (dd, 1H, H-3), 4.14
(dd, 1H, H-6b), 3.75 (ddd, 1H, H-5), 3.52 (dd, 1H, H-4), 3.47 (dd, 1H, H-6a), 3.38 (dd, JH1,H2 3.66,
1H, H-2), 3.32 (s, 3H, OMe); 2 × 0.89, 0.85, 0.79, 0.70 (5 s, 45H, C(CH3)3), 0.15, 0.12, 0.11, 0.09,
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0.04, 0.01, −0.06, −0.08, −0.11, −0.21 (10 s, 30H, SiCH3). C NMR (CDCl3, 25 °C, 100 MHz): δ
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139.1, 138.1, 137.6, 136.1, 133.3, 132.8 (6 C-ipso), 129.0, 2 × 128.6, 2 × 128.4, 2 × 128.3, 128.1,
127.9, 2 × 127.8, 127.7, 3 × 127.4, 127.3, 2 × 126.7, 126.0, 2 × 125.7, 125.4 (22 C-Ar); Residue B':
98.7 (C-1), 82.5 (C-5), 79.3 (C-4), 77.7 (C-2), 69.6 (C-3), 63.6 (C-6); Residue B: 99.8 (C-1), 84.8
(C-3), 74.8 (C-2), 73.9 (C-5), 72.7 (PhCH2), 70.7 (NapCH2), 70.7 (C-6), 70.0 (C-4); Residue A:
101.9 (CHPh), 99.2 (C-1), 80.7 (C-4), 79.0 (C-2), 75.4 (C-3), 73.8 (PhCH2), 69.1 (C-6), 62.4 (C-5),
55.3 (OMe); 26.2, 26.0, 3 × 25.9 (5 C(CH3)3), 18.3, 3 × 18.1, 17.9 (5 C(CH3)3), −3.6, −3.8, −4.1,
−4.3, −4.5, −4.6, −4.8, −5.2, 2 × −5.3 (10 SiCH3). ESI-HRMS: [M + Na]+ m/z calc for
C81H128O16Si5Na 1519.7946, found 1519.7948.
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Methyl
2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→2)[2,3,4,6-tetra-O-tert-
butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-3-O-(2-naphthylmethyl)-4-O-tert-
butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→3)]-4,6-O-benzylidene-α-D-
glucopyranoside (P3c).
General procedure for Glycosylation: from Donor D3 (50 mg) and acceptor A3 (77 mg). The
obtained product was purified by flash column chromatography (Rf = 0.40 Pentane/EtOAc 3:1) to
1
yield tetrasaccharide P3c in 30% yield (22 mg) as a white solid. H NMR (CDCl3, 25 °C, 400
MHz): δ 7.89-7.21 (m, 17H, H-Ar); Residue B': 5.15 (d, JH1,H2 6.91, 1H, H-1), 4.10 (dd, 1H, H-3),
3.88 (ddd, 1H, H-5), 3.84 (dd, 1H, H-4), 3.78 (dd, 1H, H-6b), 3.67 (dd, 1H, H-6a), 3.57 (dd,
JH1,H2 6.91, 1H, H-2); Residue B: 5.22 (d, JH1,H2 1.86, 1H, H-1), 4.85 (d, Jgem −13.12, 1H, NapCH2),
4.77 (d, Jgem −13.12, 1H, NapCH2), 4.58 (d, Jgem −12.46, 1H, PhCH2), 4.27 (d, Jgem −12.46, 1H,
PhCH2), 4.18 (dd, JH1,H2 1.86, 1H, H-2), 4.14 (dd, 1H, H-4), 3.90 (ddd, 1H, H-5), 3.54 (dd, 1H, H-
3), 3.38 (2 dd, 2H, H-6b, H-6a); Residue A: 5.49 (s, 1H, CHPh), 4.70 (d, JH1,H2 3.69, 1H, H-1), 4.38
(dd, JH2,H3 9.21, JH3,H4 9.11, 1H, H-3), 4.23 (dd, 1H, H-6b), 3.90 (ddd, 1H, H-5), 3.69 (dd, 1H, H-
6a), 3.55 (dd, JH1,H2 3.69, JH2,H3 9.21, 1H, H-2), 3.52 (dd, JH3,H4 9.11, 1H, H-4), 3.37 (s, 3H, OMe);
Residue A': 5.00 (dd, JH3,H4 9.42, JH4,H5 10.04, 1H, H-4), 4.95 (dd, JH1,H2 7.89, JH2,H3 9.60, 1H, H-2),
4.77 (dd, JH2,H3 9.60, JH3,H4 9.42, 1H, H-3), 4.25 (d, JH1,H2 7.89, 1H, H-1), 4.00 (2 dd, 2H, H-6b, H-
6a), 2.99 (ddd, JH4,H5 10.04, 1H, H-5), 2.12, 2.05, 1.94, 1.87 (4 s, 12H, CH3); 0.92, 0.88, 2 × 0.86,
0.60 (5 s, 45H, C(CH3)3), 2 × 0.12, 0.11, 0.09, 0.08, 0.07, 0.06, 0.05, −0.12, −0.36 (10 s, 30H,
SiCH3). 13C NMR (CDCl3, 25 °C, 100 MHz): δ 139.7, 137.7, 137.2, 133.4, 132.8 (5 C-ipso), 129.5,
2 × 128.5, 3 × 128.2, 3 × 127.9, 3 × 127.7, 127.3, 126.4, 125.8, 124.8, 124.6 (17 C-Ar); Residue B':
97.2 (C-1), 83.3 (C-5), 79.6 (C-4), 77.4 (C-2), 69.8 (C-3), 63.9 (C-6); Residue B: 98.4 (C-1), 83.4
(C-3), 73.6 (C-5), 72.9 (PhCH2), 72.4 (C-2), 71.4 (C-6), 70.6 (C-4), 69.3 (NapCH2); Residue A:
102.9 (CHPh), 100.7 (C-1), 81.1 (C-4), 81.0 (C-2), 72.8 (C-3), 69.5 (C-6), 62.3 (C-5), 55.8 (OMe);
Residue A': 170.6, 170.1, 169.4, 169.1 (4 C=O), 101.1 (C-1), 73.4 (C-3), 71.5 (C-5), 71.2 (C-2),
68.3 (C-4), 61.5 (C-6), 21.6, 20.9, 20.8, 20.6 (4 CH3); 26.3, 26.1, 26.0, 2 × 25.9 (5 C(CH3)3), 18.4,
18.2, 3 × 18.0 (5 C(CH3)3), −3.3, −4.1, −4.2, −4.3, 2 × −4.4, −4.9, −5.0, 2 × −5.4 (10 SiCH3). ESI-
HRMS: [M + Na]+ m/z calc for C88H140O25Si5Na 1759.8428, found 1759.8430.
Methyl
2,3,4,6-tetra-O-tert-butyldimethylsilyl-β-D-glucopyranosyl-(1→2)-3,4-O-di-tert-
butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→3)-4,6-O-benzylidene-2-O-benzyl-α-D-
glucopyranoside (P4).
General procedure for Glycosylation: from Donor D4 (50 mg) and acceptor A2 (48 mg). The
obtained product was purified by flash column chromatography (Rf = 0.55 Tol/EtOAc 12:1) to yield
trisaccharide P4 (60 mg) in 95% yield as a colorless oil. 1H NMR (CDCl3, 25 °C, 400 MHz): δ
7.40-7.20 (m, 15H, H-Ar); Residue B': 4.97 (d, JH1,H2 6.41, 1H, H-1), 4.01 (dd, 1H, H-3), 3.82 (ddd,
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The Journal of Organic Chemistry
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1H, H-5), 3.79 (dd, 1H, H-4), 3.67 (dd, 1H, H-6b), 3.63 (dd, JH1,H2 6.41, 1H, H-2), 3.58 (dd, 1H, H-
6a); Residue B: 5.57 (d, JH1,H2 2.89, 1H, H-1), 4.58 (d, Jgem −12.76, 1H, PhCH2), 4.44 (d, Jgem
−12.76, 1H, PhCH2), 4.03 (dd, JH1,H2 2.89, 1H, H-2), 3.90 (ddd, 1H, H-5), 3.86 (dd, 1H, H-3), 3.77
(dd, 1H, H-4), 3.45 (dd, 1H, H-6b), 3.32 (dd, 1H, H-6a); Residue A: 5.35 (s, 1H, CHPh), 4.84 (d,
Jgem −12.56, 1H, PhCH2), 4.53 (d, Jgem −12.56, 1H, PhCH2), 4.36 (dd, JH2,H3 9.17, JH3,H4 9.40, 1H,
H-3), 4.33 (d, JH1,H2 3.62, 1H, H-1), 4.15 (dd, Jgem −10.21, 1H, H-6b), 3.79 (ddd, JH5,H6b 4.78, 1H,
H-5), 3.58 (dd, Jgem −10.21, 1H, H-6a), 3.42 (dd, JH1,H2 3.62, JH2,H3 9.17, 1H, H-2), 3.39 (dd, JH3,H4
9.40, 1H, H-4), 3.34 (s, 3H, OMe); 0.93, 3 × 0.88, 0.77, 0.67 (6 s, 54H, C(CH3)3), 0.22, 3 × 0.10,
0.09, 0.08, 0.06, 0.05, 0.01, −0.11, −0.13, −0.14 (12 s, 36H, SiCH3). 13C NMR (CDCl3, 25 °C, 100
MHz): δ 139.3, 138.4, 137.8 (3 C-ipso), 129.2, 2 × 128.6, 2 × 128.5, 2 × 128.4, 2 × 128.3, 128.0, 2
× 127.6, 127.3, 2 × 126.9 (15 C-Ar); Residue B': 102.6 (C-1), 82.3 (C-5), 79.7 (C-4), 77.9 (C-2),
69.9 (C-3), 64.0 (C-6); Residue B: 99.0 (C-1), 79.1 (C-2), 77.8 (C-3), 74.0 (C-5), 73.4 (C-4), 72.9
(PhCH2), 72.1 (C-6); Residue A: 102.6 (CHPh), 99.4 (C-1), 80.7 (C-4), 80.6 (C-2), 73.7 (PhCH2),
73.4 (C-3), 69.3 (C-6), 62.7 (C-5), 55.4 (OMe); 26.4, 26.3, 2 × 26.1, 2 × 26.0 (6 C(CH3)3), 18.4,
18.3, 3 × 18.1, 18.0 (6 C(CH3)3), −2.6, −3.2, −3.5, 2 × −3.6, 2 × −4.3, −4.4, −4.7, −4.8, −5.3, −5.4
(12 SiCH3). ESI-HRMS: [M + Na]+ m/z calc for C76H134O16Si6Na 1493.8185, found 1493.8189.
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Methyl
2,3,4-tri-O-tert-butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→2)-3-O-(2-
naphthylmethyl)-4-O-tert-butyldimethylsilyl-6-O-benzyl-β-D-glucopyranosyl-(1→3)-4,6-O-
benzylidene-2-O-benzyl-α-D-glucopyranoside (P5b).
General procedure for Glycosylation: from Donor D5 (50 mg) and acceptor A2 (48 mg). The
obtained product was purified by flash column chromatography (Rf = 0.65 Pentane/EtOAc 9:1) to
yield trisaccharide P5b (48 mg) in 75% yield as a white solid. 1H NMR (CDCl3, 25 °C, 400 MHz):
δ 7.82-7.14 (m, 27H, H-Ar); Residue B': 5.10 (d, JH1,H2 6.04, 1H, H-1), 4.35 (d, 1H, PhCH2), 3.99
(dd, 1H, H-4), 3.89 (dd, 1H, H-3), 3.78 (ddd, 1H, H-5), 3.69 (dd, JH1,H2 6.04, 1H, H-2), 3.59 (dd,
1H, H-6b), 3.49 (dd, 1H, H-6a); Residue B: 5.42 (d, JH1,H2 3.18, 1H, H-1), 4.94 (d, Jgem −11.73, 1H,
NapCH2), 4.65 (d, Jgem −11.73, 1H, NapCH2), 4.56 (d, Jgem −12.47, 1H, PhCH2), 4.34 (d, Jgem
−12.47, 1H, PhCH2), 4.22 (dd, JH1,H2 3.18, 1H, H-2), 4.14 (dd, 1H, H-4), 3.79 (ddd, 1H, H-5), 3.58
(dd, 1H, H-3), 3.49 (2 dd, 2H, H-6b, H-6a); Residue A: 5.37 (s, 1H, CHPh), 4.68 (d, Jgem −12.38,
1H, PhCH2), 4.42 (d, Jgem −12.38, 1H, PhCH2), 4.37 (d, JH1,H2 3.55, 1H, H-1), 4.30 (dd, JH2,H3 9.19,
JH3,H4 9.14, 1H, H-3), 4.15 (dd, 1H, H-6b), 3.77 (ddd, 1H, H-5), 3.52 (dd, JH3,H4 9.14, 1H, H-4), 3.52
(dd, 1H, H-6a), 3.42 (dd, JH1,H2 3.55, JH2,H3 9.19, 1H, H-2), 3.31 (s, 3H, OMe); 0.88, 0.87, 0.83, 0.70
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(4 s, 36H, C(CH3)3), 0.13, 2 × 0.09, 0.04, 0.01, −0.02, −0.09, −0.22 (8 s, 24H, SiCH3). C NMR
(CDCl3, 25 °C, 100 MHz): δ 139.2, 138.5, 138.4, 137.8, 136.4, 133.4, 132.9 (7 C-ipso), 129.1, 2 ×
128.5, 2 × 128.4, 5 × 128.3, 128.0, 127.9, 2 × 127.8, 5 × 127.6, 127.4, 127.3, 2 × 126.8, 126.0,
125.8, 125.7, 125.6 (27 C-Ar); Residue B': 99.7 (C-1), 80.6 (C-4), 78.9 (C-5), 77.4 (C-2), 73.2
(PhCH2), 71.4 (C-6), 70.8 (C-3); Residue B: 99.8 (C-1), 85.0 (C-3), 75.5 (C-2), 74.2 (C-5), 72.9
(PhCH2), 71.0 (C-6), 70.9 (NapCH2), 70.3 (C-4); Residue A: 102.1 (CHPh), 99.2 (C-1), 80.8 (C-4),
79.9 (C-2), 75.2 (C-3), 73.8 (PhCH2), 69.2 (C-6), 62.6 (C-5), 55.3 (OMe); 26.2, 3 × 26.0 (4
C(CH3)3), 18.2, 2 × 18.1, 18.0 (4 C(CH3)3), −3.7, −3.8, −4.0, −4.3, −4.4, −4.6, −4.8, −5.0 (8 SiCH3).
ESI-HRMS: [M + Na]+ m/z calc for C82H120O16Si4Na 1495.7551, found 1495.7549.
Methyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→2)[2,3,4-tri-O-tert-butyldimethylsilyl-6-
O-benzyl-β-D-glucopyranosyl-(1→2)-3-O-(2-naphthylmethyl)-4-O-tert-butyldimethylsilyl-6-O-
benzyl-β-D-glucopyranosyl-(1→3)]-4,6-O-benzylidene-α-D-glucopyranoside (P5c).
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