Bivalent glycoconjugates based on 1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (“bimane”) as a central scaffold

Article abstract of DOI:10.1016/j.carres.2019.01.002

The heteroaromatic fused diazabicyclic “bimane” ring system, discovered four decades ago, is endowed with remarkable chemical and photophysical properties. No carbohydrate derivatives of bimanes have, however, been described thus far. Here we report on the syntheses of a range of bimanes decorated with various glycosyl residues. Mono- and disaccharide residues were attached to syn- or anti-bimane central cores via thio-, disulfido- or selenoglycosidic linkages to obtain novel fluorescent or nonfluorescent glycoconjugates. Cu(I)-catalyzed cycloaddition of glycosyl azides to a bimane diethynyl derivative furnished further bivalent glycoconjugates with sugar residues linked to the central bimane core via 1,2,3-triazole rings. We have determined the crystal and molecular structures of several glycosylated and non-glycosylated bimanes and report fluorescence data for the new compounds.

Full text of DOI:10.1016/j.carres.2019.01.002

Accepted Manuscript
Bivalent glycoconjugates based on 1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
(“bimane”) as a central scaffold
Tamás Szabó, Attila Bényei, László Szilágyi
PII:
S0008-6215(18)30609-8
https://doi.org/10.1016/j.carres.2019.01.002
CAR 7661
DOI:
Reference:
To appear in: Carbohydrate Research
Received Date: 18 October 2018
Revised Date: 7 December 2018
Accepted Date: 5 January 2019
Please cite this article as: Tamá. Szabó, A. Bényei, Láó. Szilágyi, Bivalent glycoconjugates based on
1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (“bimane”) as a central scaffold, Carbohydrate Research
(2019), doi: https://doi.org/10.1016/j.carres.2019.01.002.
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ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
Bivalent glycoconjugates based on 1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
(“bimane”) as a central scaffold.
a
b
c*
Tamás Szabó , Attila Bényei and László Szilágyi
a
b
c
Richter Gedeon Nyrt. Budapest, Department of Physical Chemistry and Department of
Organic Chemistry,University of Debrecen, H-4032 Debrecen, Egyetem tér 1. Hungary.
lszilagyi@unideb.hu
Keywords:
heteroaromatics, fluorescence, thioglycoside, selenoglycoside, disulfide,
1
,2,3-triazole, X-ray structures
Declerations of interest: none.
1

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Abstract
The heteroaromatic fused diazabicyclic “bimane” ring system, discovered four
decades ago, is endowed with remarkable chemical and photophysical properties. No
carbohydrate derivatives of bimanes have, however,been described thus far. Here we report
on the syntheses of a range of bimanes decorated with various glycosyl residues. Mono- and
disaccharide residues were attached to syn- or anti-bimane central cores via thio-, disulfido-
or selenoglycosidic linkages to obtain novel fluorescent or nonfluorescent glycoconjugates.
Cu(I)-catalyzed cycloaddition of glycosyl azides to a bimane diethynyl derivative furnished
further bivalent glycoconjugates with sugar residues linked to the central bimane core via
1
,2,3-triazole rings. We have determined the crystal and molecular structures of several
glycosylated and non-glycosylated bimanes and report fluorescence data for the new
compounds.
1
. Introduction
Cell surface-presented glycans are targets of molecular recognition events in a wide
array of physiological processes. Binding of proteins with carbohydrate-recognition domains,
such as lectins, to glycan chains anchored to the cell wall play pivotal roles in intracellular
signaling, cell-cell-, or cell-extracellular matrix interactions (for recent reviews see references
[
1,2]). Small molecular weight oligo/multivalent glycoconjugates were often found
instrumental to model the glycan partner in experimental settings under in vitro conditions
3]. Compounds displaying promising activity in these preliminary tests then qualify them to
[
be carried over to further assays of increasing physiological relevance such as inhibition of
lectin binding to cultured cells or histochemical assays in fixed animal tissue sections [4].
Although lectins are exempt of enzymic activity, as a rule (exceptions exist), O-glycosidic
bonds in lectin-directed glycoconjugates are, however, frequently replaced by non-
hydrolyzable (thio-, disulfido- or selenoglycosidic) linkages. This may be regarded as a
prudential measure to avoid inhibitor deactivation by glycan hydrolases or other carbohydrate
processing enzymes that may be present in complex biological systems used for testing lectin
inhibition activities.
A novel class of carbohydrate structures designed along this principle, diglycosyl
disulfides and disulfidoglycosides (for a review see reference [5]), received attention recently
for their various biological activities. Symmetric diglycosyl disulfides, as one of the first
platforms in this line, were found to display lectin inhibition activities in solid phase tests and
in tumor cell lines [6,7]. Some oligovalent derivatives featuring mannosyl moieties attached
2

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to benzene central cores by disulfide linkages were shown to bind to concanavalin A with
affinities surpassing that of the cognate sugar [8]. Remarkable inhibitory activities against
Trypanosoma cruzi, the etiologic agent of Chagas’s disease, were recorded in vitro for similar
structures with galacto configurations [9]. Further glycoconjugates with galactose or lactose
moieties attached to central naphthalene scaffolds proved to inhibit the binding of a plant
agglutinin and different human galectins to tumor cell lines, this bioactivity being preserved
when tested in animal tissue sections as well [4]. Exploring the scope for potential
th
biorelevance of sugar mimetics with the glycosidic oxygen substituted with another VI
column heteroatom, selenium, di-β-D-galactopyranosyl selenide and -diselenide have been
identified to display lectin inhibition efficiency comparable to that of their thio analogs [10].
On the other hand, we have recently observed significant growth inhibiton efficiency of
symmetric diglycosyl diselenides against T. brucei species, the parasite causing African
sleeping sickness [11]. As a further extension of the palette of three-bond interglyosidic
connections with two non-oxygen heteroatoms (for a review see reference [5]) we have
recently introduced the disulfide analog selenosulfide linkage which is of rare occurrence in
carbohydrate chemistry [12].
Building upon various biological activities discussed above we have set out to explore
novel glycoconjugates with a central heteroaromatic core called “bimane”. Here we wish to
report on the syntheses and properties of bivalent glycosyl derivatives based on this
remarkable heterocyclic skeleton. The beautifully symmetric heterocyclic ring system, 1,5-
diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (bimane) was discovered, after some preliminary
attempts, by E.M. Kosower et al. in 1978 [13]; an alternative synthesis being reported just
recently [14]. Bimanes are fluorescent molecules, the syn isomers displaying much stronger
effects than the anti forms (see Chart 1) [15]. The latter show, on the other hand, low
temperature phosphorescence [16,17].
Chart 1. Syn and anti forms of substituted bimanes labeled using the shorthand notation
introduced by Kosower et al. [13]
3

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Bimane fluorophores are extensively used in biophysical studies such as for
monitoring conformational changes, motions and interactions in proteins [18-21]. We have
envisioned that glycoconjugates endowed with fluorescent properties might be further
exploited to explore potential interactions with cells or higher organisms. (see, e.g. references
[
22,23]). Another motivation for our studies was provided by the observation of antiparasitic
activity of certain bimane derivatives [24]. Finally, while sugars are vital components of all
biosystems and displaying a large variety of biological activities, to some surprise, no
carbohydrate derivatives of bimanes have been described thus far to the best of our
knowledge.
Haloalkyl-bimanes like the syn and anti forms of dibromobimane (DBB) derivatives
1
a and 1b [15] (Chart 2) are reactive species and reactions with thiols are of interest, in
particular, for fluorescent labeling in biological systems [18-21].
Chart 2. Structures of syn-(CH Br,CH )B (1a) and anti-(CH Br,CH )B (1b)
2
3
2
3
Bis-thioether derivatives were obtained in smooth reactions of DBB with aliphatic/aromatic
thiols, as well as with cysteine [22], substituted cysteines [25] or glutathione [22,26].
Formation of thia-bridged derivatives (Chart 3) as minor side products have also been
observed in the reactions with thiols [22,27], with sodium sulfide [27] or H S [22].
2
Chart 3. Bimane thioethers described previously
4

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These reactions have been proposed for the purpose of quantification of glutathione [28],
sulfide- or thiol levels or H S [22] in physiological concentrations. It is to be noticed that
2
reaction of monochlorobimane (syn-(CH Cl,Me)B) with selenocysteine did not result in the
2
expected selenoether derivative, syn-(Me,Me)bimane was obtained instead via reduction of
the CH Cl group by Se-cysteine [29].
2
2
. Results and Discussion
Dibromobimanes 1a and 1b (Chart 2) were found to be suitable starting compounds
for the attachment of glycosyl residues to the bimane skeleton. First we have explored
reactions of 1a and 1b with per-O-acetylated glycosyl thiols 4a,b,αc,βc,d and observed the
formation of bis-thioglycosylated bimanes in syn (8a,b,αc,βc,d) and anti forms
(7a,b,αc,βc,d), respectively, under mild conditions and in good yields (Scheme 1 and Table
1
). Reactions with syn- and anti DBB (1a, 1b) occurred with retention of the anomeric
configurations of the starting thiols to provide β-glycosylated derivatives as ascertained
trivially by the JH1,H2 values for the gluco- (7a, 8a), galacto- (7b, 8b) or lacto (7d, 8d)
configurations (see Experimental). Availability of mannosyl thiols in both anomeric
configurations, α (4αc) and β (4βc) [30], enabled the preparation of both α and β
thioglycosides (7,8αc and 7,8βc, respectively). In these cases the anomeric configurations
were deduced from NOESY spectra wherein the presence of crosspeaks between H1/H3 and
H1/H5 of the mannosyl moiety clearly indicated β configurations for 7,8βc whereas such
1
crosspeaks were missing from the spectra of 7,8αc. H chemical shifts of H-5 were,
furthermore, found highly diagnostic for the assignment of anomeric configurations of
thiomannosides as values for α-anomers are ~ 0.5 ppm larger than those measured in the β-
counterparts (see Experimental). Selenoglycosides cannot be prepared in analogy with
glycosyl thiols because of the known instability of the corresponding glycosyl selenols [31].
On the other hand, selenoglycosides 6a,b and 5a,b,c, respectively, were readily obtained from
1
a, 1b using per-O-acetylated glycosyl isoselenuronium salts 3a,b,c as glycosyl-selenenol
equivalents following a procedure we have previously described [32] (Scheme 1 and Table 1).
5

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Scheme 1. Syntheses of glycosylated bimanes with thio- or selenoglycoside linkages
In view of the versatile activities of carbohydrate molecules with disulfide glycosidic
linkages observed in interactions with systems of increasing biorelevance [4] we have set out
to prepare glycosylated bimanes featuring this type of glycosidic bond. Of the several
methods/approaches available to synthesize unsymmetrical disulfides in general [33] and in
sugar chemistry, in particular [5,34-37], we have chosen to take advantage of the
glycosylsulfenyl-transfer properties of glycosyl sulfenamides (9) in reactions with thiols [36].
Indeed, reactions of syn-(CH SH,CH )B (1d) with the appropriate N-phthalyl-S-(2,3,4,6-
2
3
tetra-O-acetyl-β-D-glycopyranosyl)sulfonamides [36] furnished the desired
bis(glycopyranosyl-dithio) bimane derivatives 10a-c (Scheme 2, Table 1).
Scheme 2. Syntheses of bis(glycopyranosyl-dithio)bimanes
The required syn-(CH SH,CH )B (1d) was obtained from 1a via reaction with potassium
2
3
thiolacetate followed by acidic deacetylation. No anti-thiol (1g) could, however, be obtained
6

ACCEPTED MANUSCRIPT
from 1b as treatment of the thiolacetate 1f with HCl/MeOH resulted in, surprisingly, total
decomposition with the formation of polymeric products. Attempted deacetylation of 1f under
basic conditions (ammonia, LiOH) led to similar result. On the other hand, we have observed
the formation of internal disulfide 1e upon oxidation of 1d with H O (Scheme 3, Table 1).
2
2
Scheme 3. Reactions to obtain bimane thiols from DBB
As a further option to attach glycosyl moieties to the bimane skeleton we have
explored approaches by taking advantage of the Cu(I)-catalyzed azide-alkyne cycloaddition
(CuAAC, “click” ) reaction (for a review of applications in carbohydrate chemistry, see
reference [38]) for this purpose. We have selected the bimane diacetylene derivative 1h [39]
for reactions with glycosyl azides 12a,b,c,d under “click” conditions and, to our satisfaction,
3
,7-disubstituted bimane triazole derivatives 13a,b,c,d could be obtained in moderate to good
yields (Scheme 4 and Table 1).
Scheme 4. Syntheses of glycosylated bimane triazole derivatives
7

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Table 1. List of the new synthesized compounds
Structure*
Entry Compound
Yield
%)
syn / anti (R ,R )B
2
1
(
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
.
.
.
.
.
.
.
.
1c
1d
1e
1f
5a
5b
5c
6a
6b
7a
7b
7βc
7αc
7d
8a
8b
8βc
8αc
8d
10a
10b
10βc
13a
13b
13c
13d
syn
syn
syn
anti
anti
anti
anti
syn
syn
anti
anti
anti
anti
anti
syn
syn
syn
syn
syn
syn
syn
syn
syn
syn
syn
syn
(CH SAc,CH )B
(CH SH,CH )B
2 3
(CH S*,CH )B
2 3
(CH SAc,CH )B
2 3
(CH Se-β-GlcAc ,CH )B
2 4 3
(CH Se-β-GalAc ,CH )B
2 4 3
(CH Se-β-ManAc ,CH )B
2 4 3
(CH Se-β-GlcAc ,CH )B
2 4 3
(CH Se-β-GalAc ,CH )B
2 4 3
(CH S-β-GlcAc ,CH )B
2 4 3
(CH S-β-GalAc ,CH )B
2 4 3
(CH S-β-ManAc ,CH )B
2 4 3
62
87
63
85
83
59
64
62
68
80
84
98
77
95
86
77
73
21
90
81
71
11
97
57
60
57
2
3
.
0.
1.
2.
3.
4.
5.
6.
7.
8.
9.
0.
1.
2.
3.
4.
5.
6.
(CH S-α-ManAc ,CH )B
2
4
3
(CH S-β-LacAc ,CH )B
2
7
3
(CH S-β-GlcAc ,CH )B
2 4 3
(CH S-β-GalAc ,CH )B
2 4 3
(CH S-β-ManAc ,CH )B
2 4 3
(CH S-α-ManAc ,CH )B
2
4
3
(CH S-β-LacAc ,CH )B
2
7
3
(CH S -β-GlcAc ,CH )B
2 2 4 3
(CH S -β-GalAc ,CH )B
2
2
4
3
(CH S -β-ManAc ,CH )B
2
2
4
3
(CH ,TA-β-GlcAc )B
3
4
(CH ,TA-β-GalAc )B
3
4
(CH ,TA-β-ManAc )B
3
4
(CH ,TA-β-LacAc )B
3
7
*
For the notation see Experimental
Of the new compounds the structures of glycosylated and non-glycosylated derivatives have
been confirmed by crystallographic analyses. For ORTEP views of structures 1c and 1d
(Figure S1) as well as for details of structure determination and refinement see Supplementary
Information (Table S1). 1e (Figure 1) represents the first example of a bimane structure with
an intramolecular disulfide incorporated into a seven-membered ring anellated to the bimane
skeleton.
8

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Figure 1. ORTEP View of 1e at 50% probability level.
In case of 7a (Figure 2) one half of the molecule occupies the asymmetric unit and the other
half is given by a twofold screw axis. For 1c, 1e and 8a (Figure 3) two independent molecules
could be found in the asymmetric unit with small conformational differences (Figure S2).
Figure 4 shows the molecular structure of triazole derivative 13c.
Figure 2. ORTEP view of 7a at 30% probability level with partial numbering scheme.
Hydrogen atoms are omitted for clarity. Symmetry code (i): -y+1, -x+1, -z+3/2.
9

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Figure 3. ORTEP view of 8a with partial numbering scheme at 50 % probability level.
Hydrogen atoms are omitted for clarity. Only one of the molecules in the asymmetric unit is
shown.
Figure 4. ORTEP view of 13c at 50% probability level with partial numbering scheme.
Hydrogen atoms are omitted for clarity.
Distances between glycosyl units in similar diglycosylated naphthalene derivatives have been
shown to matter in sugar-lectin interactions [4]. Compounds 7a and 13c represent increasing
interglycosidic separations as measured by the through-space distances between the anomeric
C-atoms from the crystal structures (Figures 5 & 6).
1
0

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Figure 5. Interglycosidic separation distance (in Ǻ) in 7a.
Figure 6. Interglycosidic separation distance (in Ǻ) in 13c.
Search of the Cambridge Structural Database [40] (Ver. 5.39 updates May, 2018) resulted 28
hits for syn- and 11 hits for anti bimane structures. Comparison of the bond distance and bond
angle data found in the CSD reveals that the corresponding data in our compounds are in the
expected range (Table S2). The bimane structures found in the CSD also revealed
nonplanarity in several cases, i.e. angles > 0 deg. subtended by the mean planes of the two
anellated pyrazolone rings, values being varied between 0 and 50 deg. In the present cases we
have often detected two independent molecules in the asymmetric unit of the crystal. It is of
note that the extent of nonplanarity within these pairs varied significantly, for example, 13 /
2
2 deg. in 1c or 5 / 19 deg. in 8a. Even for 1e where the two rings are bridged by a disulfide
linkage the respective data are 4.4 / 18.8 deg. (Figure S2 and Table S3). Moreover, this type
of bending occurs in both syn- (8a) and anti (7a) bimanes (Table S3). These results suggest
that the planarity of the bimane backbone is not as prevalent, as it was suggested in early
single crystal studies [41, 42]. Further investigations of the same authors had shown the
1
1

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flexibility of the bimane system [43, 44]. Scatter plot of the N-N distance versus interplanar
angles revealed no correlation between these structural descriptors. Should electronic effects,
such as overlap of orbitals, govern the interplanar bending, these effects are expected to be
very similar for two molecules occupying the same asymmetric unit. However, this is not the
case as noted above. We therefore suggest that differences in the deviations from planarity for
chemically identical molecules may arise from lattice (packing) constraints in the solid state
in these particular structures.
Bimanes are fluorescent compounds with the syn isomers displaying distinctly
stronger effect than their anti counterparts as noted above. As this photophysical property is
influenced not only by syn/anti relationships but other structural subtleties as well [45,46], it
was of interest to investigate the effects, if any, of the pending glycosyl moieties on the
fluorescent behavior of these molecules. First, we note that absorption and fluorescence
spectra of compounds with the same glycosidic bond type but bearing different sugar moieties
(eg. 8a,b,c,d, see Figures S4 & S7) are practically superimposable therefore data for glucosyl
derivatives have only been listed in Table 2. Clearly, the sugar part has no effect on the
optical absorption and fluorescence properties of these molecules with absorption/ emission
maxima and extinction coefficients similar to those published for non-glycosylated bimane
derivatives [15,47]. Data for anti-bimane derivatives are not listed either because, in line with
literature reports [15,47], these isomers are non-fluorescent. It is of note, however, that the syn
selenoglycoside 6a was found nonfluorescent unlike its thio-analog 8a. It is known, however,
that when “Se is connected to the fluorophore ( …) the efficient photoinduced eletron-transfer
(PET) process between the Se and fluorophore quenches the fluorescence” [48]. On the other
hand, this behavior may also be related to small value of the extinction coefficient (800) for
the band used for fluorescent excitation in 6a in comparison with significantly higher values
in other syn-bimanes (4 ~ 5000, Table 2).
1
2

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Table 2. Ultraviolet-visible absorption and emission properties of syn-bimanes
[
syn-(R ,R )B]*
2 1
Compd. R2
No.
R1
Absorbance
Excitation Emission
Quantum
yield Φ F
0.86
0.70
0.74
-1 .
-1
λabs (nm) / ε (M cm )
375 / 5600, 238 / 21100
375 / 5400, 237 / 14300
360 /4200, 232 / 13100
360 / 800, 237 / 26100
364 / 5100, 255 / 23100
378 / 4600, 245 / 13200
370 / 3500, 242 / 4400
λex (nm)
λem (nm)
1
1
1
6
8
1
1
c
CH
2
2
2
2
2
2
3
SAc
SH
S-
CH
CH
CH
CH
CH
CH
3
3
3
3
3
3
370
445
d
e
a
a
0a
3a
CH
CH
CH
CH
CH
CH
370
445
370
442
Se-β-GlcAc
S-β-GlcAc
-β-GlcAc
370
n.d.
-
4
370
444
0.89
0.57
0.86
4
S
370
449
2
4
TA-β-
370
475
GlcAc4
*
For the notation, see Experimental
3
. Conclusion
In summary we have described the syntheses and characterization of a set of novel
derivatives, including those with appended mono- and disaccharide moieties, based on the
,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (bimane) heteroaromatic ring system. Mono-
1
and disaccharide residues were attached to syn- or anti-bimane central cores via thio-, dithio-
or selenoglycosidic linkages to obtain novel fluorescent or nonfluorescent glycoconjugates.
Cu(I)-catalyzed cycloaddition of glycosyl azides to a bimane diethinyl derivative furnished
further bivalent glycoconjugates with sugar residues linked to the central bimane core via
1
,2,3-triazole rings. We have determined the crystal and molecular structures of several
glycosylated and non-glycosylated bimanes and report fluorescence data for the new
compounds.
4
. Experimental
.1. General information
4
syn-(CH Br,CH )B, anti-(CH Br,CH )B [15], syn-(CH ,C≡CTMS)B [39], N-phthalyl-S-
2
3
2
3
3
(
2,3,4,6-tetra-O-acetyl-β-D-glycopyranosyl)sulfonamides [36], per-O-acetylated-β-D-
glycopyranosyl azides [49], 2,3,4,6-Tetra-O-acetyl-β- -glycopyranosyl isoselenuronium
bromides [31,32], 1-thio-2,3,4,6-tetra-O-acetyl-β- -glycopyranoses [30,50-52] and 1-thio-
D
D
2
,3,6-tri-O-acetyl-4-(2,3,4,6-tetra-O-acetyl-β-
D
-galactopyranosyl)-β-D-glucopyranose [53]
1
3

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were prepared according to literature procedures. TLC was performed on DC-Alurolle
Kieselgel F254 (Merck), and the spots on the plates visualized under UV light (254 nm) and
by heating. For column chromatography Kieselgel 60 (Merck, particle size 0.063-0.200) was
1
13
used. NMR spectra were recorded on a Bruker Avance II 500 (500/125 MHz for H/ C)
spectrometer. Chemical shifts measured in ppm are referenced to internal Me Si (0.00 ppm
4
1
13
for H), or to the residual solvent signals (CDCl : 77.23 ppm, DMSO-d : 39.51 ppm for C).
3
6
Mass spectra were measured using Bruker microOTOF-Q or Thermo LTQ FT Ultra
spectrometers. UV and steady state fluorescence data were determined on a Perkin Elmer
Lambda 11 and Jasco FP-8200 spectrometer using 1 µmol solutions in MeCN in a quartz
cuvette of 10 mm length The excitation and emission spectra were recorded using 2.5 nm
excitation, 5.0 nm emission bandwith and 200 nm/min scanning speed. Quantum yields (Φ)
were calculated according to the equation:
F (ε c ΦF )
s
ref ref
ref
ΦF =
s
F (ε c )
ref
s s
using syn-(CH ,CH )B as reference (ΦF = 0.72) [15]. Symbols: F: integrated area under
3
3
ref
fluorescence curve, s: sample, ref: syn-(CH ,CH )B, c: concentration, ε: absorption
3
3
coefficient.
X-ray diffraction data were collected at 293-298 K using a Bruker-D8 Venture diffractometer
equipped with INCOATEC IµS 3.0 dual (Cu and Mo) sealed tube microsources and Photon 2
Charge-integrated Pixel Array detector. For compounds 1c, 1d, 1e and 13c Mo Kα (λ =
0
.7107 Å) while for 7a and 8a Cu Kα (λ = 1.541 Å) radiation was applied. For the software
used for data collection and processing see Supporting Information. The structures could be
2
solved using direct methods and refined on F using SHELXL program [54] incorporated into
the APEX3 suite. Refinement was performed anisotropicaly for all non-hydrogen atoms.
1
4

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Hydrogen atoms atoms were placed into geometric positions except the SH protons in 1d as
these hydrogen atoms could be found at the difference electron density map. Tables were
extracted from the edited CIF file using publCIF [55]. The PLATON program [56] was used
for crystallographic calculations. Further information on the data collection and refinement
for the respective compounds can be found in Table S1. CCDC numbers for compounds 1c,
1
d, 1e, 7a, 8a, 13c are 1847474-1847479, respectively. The absolute configurations for
compounds 7a, 8a and 13c were determined on the basis of the configuration of stereogenic
centers in the carbohydrate moiety.
4
.2.
syn-(CH SAc,CH )B, 4,6-bis-[(acetylthio)methyl]-3,7-dimethyl-1,5-
2 3
diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (1c)
To a solution of 1a (3.50 g, 10.0 mmol) in CH Cl (50 mL) potassium thioacetate
2
2
(3.43 g, 30.0 mmol) was added. The suspension was stirred at room temperature for 1.5 h,
then filtered through silica gel (2.00 g), and washed with CH Cl (50 mL). The filtrate was
2
2
evaporated to dryness and the residue crystallized from MeOH, to yield 1c (2.10 g, 62 %) as
1
yellow needles. R 0.48 (9:1 CHCl -EtOAc), m.p. 131-135 °C, H NMR (DMSO-d , 500
f
3
6
1
3
MHz): δ (ppm) 4.25 (s, 4H, 2xCH ), 2.43 (s, 6H, 2xCH ), 1.78 (s, 6H, 2xCH ), C NMR
2
3
3
(
DMSO-d , 125 MHz): δ (ppm) 193.4 (SCO), 159.6 (C-2, C-8), 146.9 (C-4, C-6), 114.2 (C-3,
6
+
C-7), 30.1 (COCH ), 22.4 (CH ), 6.8 (CH ). HRMS m/z calc. for C H N O S [M+H] :
3
2
3
14 16
2
4 2
3
41.062 Found: 341.062.
4
.3.
syn-(CH SH,CH )B, 4,6-bis-(mercaptomethyl)-3,7-dimethyl-1,5-
2 3
diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (1d)
To a cold (0 °C) mixture of MeOH (65 mL) and acetylchloride (2.6 mL, 36.4 mmol)
1
c (2.60 g, 7.6 mmol) was added. The mixture was allowed to warm to room temperature and
1
5

ACCEPTED MANUSCRIPT
stirred overnight. The precipitated solid was filtered and washed with and recrystallized from
MeOH, to yield 1d (1.70 g, 87 %) as yellow needles. R 0.17 (4:1 CHCl -MeOH), m.p. 169-
f
3
1
1
1
72 °C, H NMR (DMSO-d , 500 MHz): δ (ppm) 3.88 (s, 4H, 2xCH ), 3.75 (s, 2H, 2xSH),
6 2
1
3
.79 (s, 6H, 2xCH ), C NMR (DMSO-d6, 125 MHz): δ (ppm) 160.2 (C-2, C-8), 150.0 (C-4,
3
+
C-6), 111.7 (C-3, C-7), 17.1 (CH ), 6.4 (CH ). HRMS m/z calc. for C H N O S [M+H] :
2
3
10 12
2
2 2
2
57.041 Found: 257.041.
4
3
.4. syn-(CH S,CH )B, 4,6-(dithiamethylene)-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-
2 3
,6-diene-2,8-dione (1e)
To the mixture of MeOH (100 mL) and CH Cl (50 mL) 1d (500 mg, 0.51 mmol),
2
2
then 30% aq. H O (3.6 mL) was added. The solution was stirred at room temperature for 2 h,
2
2
then the reaction mixture was poured into a solution of 10 % NaHSO in water (200 mL) and
3
the water phase extracted with EtOAc (200 mL). The organic phase was washed with water,
dried (Na SO ) and evaporated. The crude product was purified by column chromatography
2
4
(EtOAc) to yield 1e (310 mg, 63 %) as a yellow solid. Crystals for X-ray diffraction were
1
grown from a solution in THF overlayered with diisopropylether. M.p. 151-157 °C, H NMR
1
3
(
CDCl , 500 MHz): δ (ppm) 4.12 (s, 4H, 2xCH ), 1.81 (s, 6H, 2xCH ), C NMR (CDCl , 125
3 2 3 3
MHz): δ (ppm) 159.0 (C-2, C-8), 149.4 (C-4, C-6), 113.1 (C-3, C-7), 36.1 (CH ), 6.5 (CH ).
2
3
+
HRMS m/z calc. for C H N O S [M+Na] : 277.008 Found: 277.007.
1
0
10
2
2 2
4
.5. anti-(CH SAc,CH )B, 4,8-bis-[(acetylthio)methyl]-3,7-dimethyl-1,5-
2 3
diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione (1f)
To a solution of 1b (800 mg, 2.3 mmol) in CH Cl (100 mL), potassium thioacetate
2
2
(784 mg, 6.9 mmol) was added. The suspension was stirred at room temperature for 1.5 h,
filtered through silica gel (2.00 g), and washed with CH Cl (50 mL). The filtrate was
2
2
1
6

ACCEPTED MANUSCRIPT
evaporated, and the residue crystallized from MeOH, to yield 1f (660 mg, 85 %) as a white
1
solid. R 0.64 (4:1 toluene-isopropanol), H NMR (ppm) (CDCl , 500 MHz): δ 4.19 (s, 4H,
f
3
13
2
xCH ), 2.35 (s, 6H, 2xCH ), 1.93 (s, 6H, 2xCH ), C NMR (DMSO-d , 125 MHz): δ (ppm)
2 3 3 6
1
94.0 (SCO), 160.3 (C-2, C-6), 143.9 (C-4, C-8), 114.3 (C-3, C-7), 30.0 (COCH ), 22.1
3
+
(
CH ), 6.7 (CH ). HRMS m/z calc. for C H N O S [M+Na] : 363.045 Found: 363.044.
2
3
14 16
2
4 2
4
.6. General method A for the preparation of seleno- and thio-glycosides
Compound 1a or 1b and the appropriate 3(a-c), 4(a-d) were dissolved in DMF with
stirring under nitrogen atmosphere at room temperature and triethylamine added through a
septum. Stirring was continued at room temperature for 30 min then the reaction mixture was
poured into water. A solid deposited, which was filtered, rinsed with water and dried. The
crude products were purified by column chromatography (9.5:0.5 toluene-isopropanol) to give
5
(a-c), 6(a,b) selenoglycosides and 7(a-d), 8(a-d) thioglycosides.
4
.7. anti-(CH Se-β-GlcAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2
4
3
glucopyranosyl)seleno]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione
5a)
(
According to general method A, the reaction of 1b (100 mg, 0.29 mmol) with 3a (313
mg, 0.58 mmol) and triethylamine (0.2 mL, 1.4 mmol) gave 5a (239 mg, 83 %) as a white
2
D
7
1
solid. R 0.52 (4:1 toluene-isopropanol), [α] – 5 (c 0.25 CDCl ). H NMR (CDCl , 500
f
3
3
Glc
Glc
Glc
MHz): δ (ppm) 5.23 (t, 2H, H-2,2’ , J = J2,3 = 9.5 Hz), 5.10 (m, 6H, H-3,3’ , H-4,4’ ,
1
,2
Glc
Glc
Glc
H-1,1’ ), 4.29 (dd, 2H, H-6a,6a’ , J = 4.5 Hz, J = 12.5 Hz), 4.14 (m, 4H, H-6b,6b’ ,
5
,6a
a,b
Bim
Bim
Glc
CH2a,a’ , JCH2a,b = 13.0 Hz), 3.93 (d, 2H, CH2b,b’ ), 3.77 (m, 2H, H-5,5’ ), 2.09, 2.03,
Glc
Bim 13
2
.02, 2.01 (s, 24H, 8xCOCH3 ), 1.87 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ
3
Glc
Bim
Bim
(
ppm) 170.6, 170.1, 169.5 (COCH ) , 160.7 (C-2, C-6) , 147.2 (C-4, C-8) , 112.6 (C-3,
3
1
7

ACCEPTED MANUSCRIPT
Bim
Glc
Glc
Glc
Glc
Glc
C-7) , 77.3 (C-1,1’) , 76.8 (C-5,5’) , 73.5 (C-3,3’) , 70.7 (C-2,2’) , 68.0 (C-4,4’) ,
Glc
Glc
Bim
Bim
6
1.8 (C-6,6’) , 20.7, 20.6 (COCH ) , 12.6 (CH ) , 6.7 (CH3) . HRMS m/z calc. for
3 2
+
C H N O Se [M+Na] : 1035.102 Found: 1035.102.
38
48
2
20
2
4
.8.
anti-(CH Se-β-GalAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
galactopyranosyl)seleno]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-
dione (5b)
According to general method A, the reaction of 1b (200 mg, 0.57 mmol) with 3b (626
mg, 1.2 mmol) and triethylamine (0.4 mL, 2.9 mmol) gave 5b (340 mg, 59 %) as a white
2
D
7
1
solid. R 0.52 (4:1 toluene-isopropanol), [α] + 64 (c 0.25 CDCl ). H NMR (CDCl , 500
f
3
3
Gal
Gal
MHz): δ (ppm) 5.47 (d, 2H, H-4,4’ , J = 2.5 Hz), 5.26 (t, 2H, H-2,2’ , J = J2,3 = 10.0
3
,4
1,2
Gal
Gal
Gal
Gal
Bim
a,a’
Hz), 5.08 (m, 4H, H-3,3’ , H-1,1’ ), 4.12 (m, 6H, H-6a,6a’ , H-6b,6b’ , CH
), 3.99
2
Gal
Bim
b,b’
Gal
(
m, 2H, H-5,5’ , CH2
), 2.16, 2.05, 2.04, 1.99 (s, 24H, 8xCOCH3 ), 1.88 (s, 6H,
Bim 13
Gal
2
2
xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.3, 169.9, 169.8 (COCH ) , 160.6 (C-
3
3
Bim
Bim
Bim
Gal
Gal
, C-6) , 147.1 (C-4, C-8) , 112.5 (C-3, C-7) , 78.2 (C-1,1’) , 75.6 (C-5,5’) , 71.4
Gal
Gal
Gal
Gal
Gal
(
C-3,3’) , 68.0 (C-4,4’) , 67.3 (C-2,2’) , 61.1 (C-6,6’) , 20.7 (COCH ) , 12.9
3
Bim
Bim
+
(
CH2) , 6.6 (CH3) . HRMS m/z calc. for C H N O Se [M+Na] : 1035.102 Found:
38 48 2 20 2
1
035.103.
4
.9.
anti-(CH Se-β-ManAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
mannopyranosyl)seleno]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-
dione (5c)
According to general method A, the reaction of 1b (50 mg, 0.14 mmol) with 3c (157
mg, 0.29 mmol) and triethylamine (0.2 mL, 1.4 mmol) gave 5c (92 mg, 64 %) as a white
2
D
7
1
solid. R 0.52 (4:1 toluene-isopropanol), [α] – 18 (c 0.25 CDCl ). H NMR (CDCl , 500
f
3
3
1
8

ACCEPTED MANUSCRIPT
Man
Man
MHz): δ (ppm) 5.73 (s, 2H, H-1,1’ ), 5.42 (d, 2H, H-2,2’ , J2,3 = 3.5 Hz), 5.34 (t, 2H, H-
Man
Man
Man
Man
4
,4’ , J3,4 = J4,5 = 10.0 Hz), 5.23 (dd, 2H, H-3,3’ ), 4.31 (m, 4H, H-6a,6a’ , H-5,5’
,
Man
J5,6a = 5.0 Hz), 4.16 (dd, 2H, H-6b,6b’ , J5,6b = 1.9 Hz, J6a,6b 12.0 Hz), 3.96 (dd, 4H,
Bim
Man
2
2
6
3
xCH2 , JCH2a,b = 13.0 Hz), 2.17, 2.10, 2.06, 1.98 (s, 24H, 8xCOCH3 ), 1.86 (s, 6H,
Bim 13
Man
xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.5, 169.7 (COCH ) , 160.1 (C-2, C-
3
3
Bim
Bim
Bim
Man
Man
)
, 145.4 (C-4, C-8) , 113.7 (C-3, C-7) , 77.9 (C-1,1’) , 71.1 (C-5,5’) , 70.6 (C-
Man
Man
Man
Man
Man
,3’) , 69.8 (C-2,2’) , 66.1 (C-4,4’) , 62.3 (C-6,6’) , 20.9, 20.7 (COCH3) , 13.9
Bim
Bim
+
(
CH2) , 6.8 (CH3) . HRMS m/z calc. for C H N O Se [M+Na] : 1035.102 Found:
38 48 2 20 2
1
035.104.
4
.10. syn-(CH Se-β-GlcAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
glucopyranosyl)seleno]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
6a)
(
According to general method A, the reaction of 1a (200 mg, 0.57 mmol) with 3a (625
mg, 1.2 mmol) and triethylamine (0.2 mL, 1.4 mmol) yielded 6a (358 mg, 62 %) as a white
2
D
7
1
solid. R 0.47 (4:1 toluene-isopropanol), [α] – 5 (c 0.25 CDCl ). H NMR (CDCl , 500
f
3
3
Glc
Glc
MHz): δ (ppm) 5.22 (t, 2H, H-2,2’ , J = J = 9.5 Hz), 5.08 (t, 2H, H-3,3’ , J = 9.5
2
,3
1,2
3,4
Glc
Glc
Glc
Hz), 5.07 (t, 2H, H-4,4’ ), 4.81 (d, 2H, H-1,1’ ), 4.28 (dd, 2H, H-6a,6a’ , J = 5.0 Hz,
5
,6a
Glc
Bim
J6a,6b = 12.5 Hz), 4.14 (dd, 2H, H-6b,6b’ , J5,6b = 1.5 Hz), 4.08 (d, 2H, CH2a,a’ , JCH2a,b
=
Bim
Glc
1
8
1
1
2
3.0 Hz), 3.99 (d, 2H, CH2b,b’ ), 3.73 (m, 2H, H-5,5’ ), 2.11, 2.06, 2.04, 2.02, (s, 24H,
Glc
Bim 13
xCOCH3 ), 1.90 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.5, 169.9,
3
Glc
Bim
Bim
Bim
69.4 (COCH ) , 160.0 (C-2, C-8) , 146.2 (C-4, C-6) , 114.2 (C-3, C-7) , 77.4 (C-
3
Glc
Glc
Glc
Glc
Glc
Glc
,1’) , 76.8 (C-5,5’) , 73.2 (C-3,3’) , 70.3 (C-2,2’) , 67.9 (C-4,4’) , 61.9 (C-6,6’) ,
Bim
Glc
Bim
0.7, 20.5 (CH2) , 13.6 (COCH ) , 7.1 (CH ) . HRMS m/z calc. for C H N O Se
2
3
3
38 48
2
20
+
[
M+H] : 1013.120 Found: 1013.123.
1
9

ACCEPTED MANUSCRIPT
4
.11. syn-(CH Se-β-GalAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2
4
3
galactopyranosyl)seleno]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-
dione (6b)
According to general method A, the reaction of 1a (100 mg, 0.29 mmol) with 3b (313
mg, 0.58 mmol) and triethylamine (0.2 mL, 1.4 mmol) furnished 6b (176 mg, 61 %) as a
27
D
1
yellow solid. R 0.47 (4:1 toluene-isopropanol), [α] – 75 (c 0.25 CDCl ). H NMR (CDCl ,
f
3
3
Gal
Gal
5
1
6
2
1
00 MHz): δ (ppm) 5.45 (d, 2H, H-4,4’ , J = 2.0 Hz), 5.24 (t, 2H, H-2,2’ , J = J2,3 =
3,4 1,2
Gal
Gal
Gal
0.0 Hz), 5.06 (dd, 2H, H-3,3’ ), 4.81 (d, 2H, H-1,1’ ), 4.11 (m, 8H, H-6a,6a’ , H-
Gal
Bim
Gal
b,6b’ , 2xCH2 ), 3.96 (t, 2H, H-5,5’ , J = J5,6b = 10.0 Hz), 2.19, 2.07, 2.06, 2.00 (s,
5,6a
Gal
Bim 13
4H, 8xCOCH3 ), 1.92 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.3,
3
Gal
Bim
Bim
Bim
70.1, 169.8 (COCH ) , 160.0 (C-2, C-8) , 146.2 (C-4, C-6) , 114.0 (C-3, C-7) , 78.3
3
Gal
Gal
Gal
Gal
Gal
(
C-1,1’) , 76.1 (C-5,5’) , 71.2 (C-3,3’) , 67.5 (C-4,4’) , 67.1 (C-2,2’) , 61.3 (C-
Gal
Gal
Bim
Bim
6
,6’) , 20.7 (COCH ) , 14.0 (CH ) , 7.1 (CH3) . HRMS m/z calc. for C H N O Se
3 2 38 48 2 20
2
+
[
M+H] : 1013.120 Found: 1013.123.
4
.12. anti-(CH S-β-GlcAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
glucopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione
7a)
(
According to general method A, the reaction of 1b (100 mg, 0.29 mmol) with 4a (219
mg, 0.60 mmol) and triethylamine (0.2 mL, 1.4 mmol) gave 7a (210 mg, 80 %) as a white
2
D
7
solid. R 0.56 (4:1 toluene-isopropanol), [α] + 6 (c 0.25 CDCl ). Crystals for X-ray
f
3
1
diffraction were grown from a solution in tBuOH overlayered with cyclohexane. H NMR
Glc
Glc
(
CDCl , 500 MHz): δ (ppm) 5.24 (t, 2H, H-2,2’ , J = J = 9.5 Hz), 5.12 (t, 2H, H-4,4’ ,
3 1,2 2,3
Glc
Glc
Glc
J3,4 = J4,5 9.5 Hz), 5.06 (t, 2H, H-3,3’ ), 4.76 (d, 2H, H-1,1’ ), 4.24 (dd, 2H, H-6a,6a’ ,
Bim
J5,6a = 4.5 Hz, J6a,6b = 10.0 Hz), 4.16 (d, 2H, CH2a,a’ , JCH2a,b = 14.5 Hz), 4.10 (dd, 2H, H-
2
0

ACCEPTED MANUSCRIPT
Glc
Bim
Glc
6
2
1
7
b,6b’ , J = 1.5 Hz), 3.91 (d, 2H, CH2b,b’ ), 3.75 (m, 2H, H-5,5’ ), 2.09, 2.04, 2.02,
5,6b
Glc
Bim 13
.01 (s, 24H, 8xCOCH3 ), 1.89 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm)
3
Glc
Bim
Bim
70.6, 170.1, 169.4 (COCH ) , 160.5 (C-2, C-6) , 145.9 (C-4, C-8) , 113.8 (C-3, C-
3
Bim
Glc
Glc
Glc
Glc
Glc
)
, 82.5 (C-1,1’) , 76.0 (C-5,5’) , 73.6 (C-3,3’) , 69.8 (C-2,2’) , 68.1 (C-4,4’) , 61.8
Glc
Bim
Glc
Bim
(
C-6,6’) , 21.7 (CH ) , 20.6 (COCH ) , 6.6 (CH ) . HRMS m/z calc. for
2 3 3
+
C H N O S [M+Na] : 939.213 Found: 939.215.
38
48
2
20 2
4
.13. anti-(CH S-β-GalAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
galactopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione
7b)
(
According to general method A, the reaction of 1b (100 mg, 0.29 mmol) with 4b (219
mg, 0.60 mmol) and triethylamine (0.2 mL, 1.4 mmol) gave 7b (220 mg, 84 %) as a white
2
D
7
1
solid. R 0.56 (4:1 toluene-isopropanol), [α] – 40 (c 0.25 CDCl ). H NMR (CDCl , 500
f
3
3
Gal
Gal
MHz): δ (ppm) 5.44 (d, 2H, H-4,4’ , J = 3.0 Hz), 5.20 (t, 2H, H-2,2’ , J = J = 10.0
3
,4
1,2
2,3
Gal
Gal
Gal
Gal
Hz), 5.07 (dd, 2H, H-3,3’ ), 4.78 (d, 2H, H-1,1’ ), 4.08 (m, 10H, H-6a,6a’ , H-6b,6b’ ,
Bim
Gal
Gal
Bim
2
xCH2 , H-5,5’ ), 2.16, 2.05, 2.04, 1.98 (s, 24H, 8xCOCH ), 1.88 (s, 6H, 2xCH3 ),
3
1
3
Gal
Bim
C NMR (CDCl , 125 MHz): δ (ppm) 170.3, 170.1, 169.6 (COCH ) , 160.4 (C-2, C-6)
,
3
3
Bim
Bim
Gal
Gal
Gal
1
6
45.7 (C-4, C-8) , 113.9 (C-3, C-7) , 83.4 (C-1,1’) , 74.5 (C-5,5’) , 71.6 (C-3,3’) ,
Gal
Gal
Gal
Bim
Gal
7.3 (C-4,4’) , (C-2,2’) , 61.0 (C-6,6’) , 22.3 (CH ) , 20.7, 20.5 (COCH ) , 6.6
2
3
Bim
+
(
CH3) . HRMS m/z calc. for C H N O S [M+Na] : 939.213 Found: 939.210.
38 48 2 20 2
4
.14. anti-(CH S-β-ManAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
mannopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione
7βc)
(
2
1

ACCEPTED MANUSCRIPT
According to general method A, the reaction of 1b (100 mg, 0.29 mmol) with 4βc
(219 mg, 0.60 mmol) and triethylamine (0.2 mL, 1.4 mmol) furnished 7βc (260 mg, 98 %) as
2
D
7
1
a white solid. R 0.56 (4:1 toluene-isopropanol), [α] + 10 (c 0.25 CDCl ). H NMR (CDCl ,
f
3
3
Man
Man
5
1
5
00 MHz): δ (ppm) 5.47 (d, 2H, H-2,2’ , J2,3 = 3.5 Hz), 5.29 (t, 2H, H-4,4’ , J3,4 = J =
4,5
Man
Man
Man
0.0 Hz), 5.10 (dd, 2H, H-3,3’ ), 5.04 (s, 2H, H-1,1’ ), 4.24 (dd, 2H, H-6a,6a’ , J5,6a
=
Man
Bim
.0 Hz, J6a,6b = 12.5 Hz), 4.15 (dd, 2H, H-6b,6b’ , J5,6b = 2.0 Hz), 4.11 (d, 2H, CH2a,a’
,
Bim
Man
JCH2a,b = 14.5 Hz), 3.95 (d, 2H, CH2b,b’ ), 3.73 (m, 2H, H-5,5’ ), 2.17, 2.09, 2.04, 1.96 (s,
Man
Bim 13
2
1
4H, 8xCOCH3 ), 1.85 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.7,
3
Man
Bim
Bim
Bim
70.0, 169.5 (COCH3) , 160.2 (C-2, C-6) , 145.4 (C-4, C-8) , 114.0 (C-3, C-7) , 81.5
Man
Man
Man
Man
Man
(
C-1,1’) , 76.5 (C-5,5’) , 71.7 (C-3,3’) , 69.9 (C-2,2’) , 65.7 (C-4,4’) , 62.6 (C-
Man
Bim
Man
Bim
6
,6’) , 22.7 (CH2) , 20.9, 20.7, 20.5 (COCH3) , 6.5 (CH3) . HRMS m/z calc. for
+
C H N O S [M+Na] : 939.213 Found: 939.214.
38
48
2
20 2
4
.15. anti-(CH S-α-ManAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-α-D-
2 4 3
mannopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione
(7αc)
According to general method A, the reaction of 1b (100 mg, 0.29 mmol) with 4αc
219 mg, 0.60 mmol) and triethylamine (0.2 mL, 1.4 mmol) furnished 7αc (201 mg, 77 %) as
(
2
D
7
1
a white solid. R 0.56 (4:1 toluene-isopropanol), [α] + 91 (c 0.25 CDCl ). H NMR (CDCl ,
f
3
3
Man
Man
Man
Man
5
00 MHz): δ (ppm) 5.30 (m, 6H, H-2,2’ , H-1,1’ , H-4,4’ ), 5.19 (dd, 2H, H-3,3’
,
Man
Man
J2,3 = 3.5 Hz, J3,4 = 10.0 Hz), 4.36 (m, 2H, H-5,5’ ), 4.26 (dd, 2H, H-6a,6a’ , J5,6a = 5.5
Man
Bim
Hz, J6a,6b = 12.5 Hz), 4.12 (dd, 2H, H-6b,6b’ , J5,6b = 2.0 Hz), 3.95 (d, 2H, CH2a,a’ , JCH2a,b
Bim
Man
=
14.0 Hz), 3.88 (d, 2H, CH2b,b’ ), 2.13, 2.07, 2.03, 1.94 (s, 24H, 8xCOCH3 ), 1.85 (s, 6H,
Bim 13
Man
2
xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.4, 169.6 (COCH ) , 159.9 (C-2, C-
3 3
Bim
Bim
Bim
Man
Man
6
)
, 143.8 (C-4, C-8) , 115.0 (C-3, C-7) , 81.5 (C-1,1’) , 70.0 (C-5,5’) , 69.4 (C-
2
2

ACCEPTED MANUSCRIPT
Man
Man
Man
Man
Bim
3
,3’) , 69.4 (C-2,2’) , 66.1 (C-4,4’) , 62.4 (C-6,6’) , 22.4 (CH2) , 20.8, 20.5
Man
Bim
+
(
COCH3) , 6.5 (CH3) . HRMS m/z calc. for C H N O S [M+Na] : 939.213 Found:
38 48 2 20 2
9
39.213.
4
.16. anti-(CH S-β-LacAc ,CH )B, 4,8-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-
D
-galactosyl-
2
7
3
(
1→4)-2,3,6-tri-O-acetyl-1-thio-β-
D
-glucosyl)thio]methyl}-3,7-dimethyl-1,5-
diazabicyclo[3.3.0]octa-3,7-diene-2,6-dione (7d)
According to general method A, the reaction of 1b (100 mg, 0.29 mmol) with 4d (392
mg, 0.60 mmol) and triethylamine (0.1 mL, 0.72 mmol) yielded 7d (407 mg, 95 %) as a white
2
D
7
1
solid. R 0.38 (4:1 toluene-isopropanol), [α] + 6 (c 0.25 CDCl ). H NMR (CDCl , 500
f
3
3
Gal
Glc
MHz): δ (ppm) 5.34 (d, 2H, H-4,4’ , J = 3.5 Hz), 5.22 (t, 2H, H-3,3’ , J = J = 9.5
3
,4
2,3
3,4
Gal
Gal
Glc
Hz), 5.09 (t, 2H, H-2,2’ , J1 = J = 8.0 Hz), 4.95 (dd, 2H, H-3,3’ ), 4.93 (t, 2H, H-2,2’ ,
,2
2,3
Glc
Gal
Glc
J1,2 = 9.5 Hz), 4.72 (d, 2H, H-1,1’ ), 4.48 (m, 4H, H-1,1’ , H-6a,6a’ , J
= 12.5 Hz),
6a,6b
Bim
Glc
Glc
Gal
Bim
4
6
2
.09 (m, 8H, CH2a,a’ , H-4,4’ , H-6b,6b’ , H-6b,6b’ ), 3.88 (m, 4H, CH
, H-
2b,b’
Gal
Gal
Glc
a,6a’ ), 3.82 (t, 2H, H-5,5’ , J = J5,6b = 10.0 Hz), 3.64 (m, 2H, H-5,5’ ), 2.14, 2.12,
5
,6a
Lac
Bim 13
.06, 2.04, 2.03, 2.02, 1.96, 1.84 (s, 42H, 14xCOCH3 , 2xCH3 ), C NMR (CDCl , 125
3
Lac
Bim
MHz): δ (ppm) 170.3, 170.0, 169.6, 169.1 (COCH ) , 160.5 (C-2, C-6) , 145.8 (C-4, C-
3
Bim
Bim
Gal
Glc
Glc
8
4
6
)
, 113.8 (C-3, C-7) , 101.0 (C-1,1’) , 82.4 (C-1,1’) , 76.7 (C-5,5’) , 75.9 (C-
Glc
Glc
Gal
Gal
Gal
Glc
,4’) , 73.6 (C-3,3’) , 71.0 (C-5,5’) , 70.7 (C-3,3’) , 70.3 (C-2,2’) , 69.1 (C-2,2’) ,
Gal
Glc
Gal
Bim
6.6 (C-4,4’) , 61.8 (C-6,6’) , 60.8 (C-6,6’) , 21.9 (CH ) , 20.9, 20.8, 20.7, 20.6, 20.5
2
Lac
Bim
+
(
COCH ) , 6.6 (CH ) . HRMS m/z calc. for C H N O S [M+Na] : 1515.382 Found:
3
3
62 80
2
36 2
1
515.390.
2
3

ACCEPTED MANUSCRIPT
4
.17. syn-(CH S-β-GlcAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2
4
3
glucopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
8a)
(
According to general method A, the reaction of 1a (100 mg, 0.29 mmol) with 4a (219
mg, 0.60 mmol) and triethylamine (0.1 mL, 0.72 mmol) yielded 8a (225 mg, 86 %) as a pale
27
D
yellow solid. R 0.61 (4:1 toluene-isopropanol), [α] – 97 (c 0.25 CDCl ). Crystals for X-ray
f
3
1
diffraction were grown from a solution in tBuOH overlayered with cyclohexane. H NMR
Glc
Glc
(
CDCl , 500 MHz): δ (ppm) 5.24 (t, 2H, H-3,3’ , J = J = 9.5 Hz), 5.08 (t, 2H, H-4,4’ ,
3 2,3 3,4
Glc
Glc
J = 9.5 Hz), 5.03 (t, 2H, H-2,2’ , J = 9.5 Hz), 4.53 (d, 2H, H-1,1’ ), 4.27 (dd, 2H, H-
4
,5
1,2
Glc
Glc
6
2
2
a,6a’ , J = 5.0 Hz, J6a,6b = 12.5 Hz), 4.13 (dd, 2H, H-6b,6b’ , J = 1.5 Hz), 4.11 (d,
5
,6a
5,6b
Bim
Bim
Glc
H, CH2a,a’ , JCH2a,b = 14.0 Hz), 4.01 (d, 2H, CH2b,b’ ), 3.73 (m, 2H, H-5,5’ ), 2.11, 2.06,
Glc
Bim 13
.04, 2.02 (s, 24H, 8xCOCH3 ), 1.94 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ
3
Glc
Bim
Bim
(
ppm) 170.5, 170.0, 169.3 (COCH ) , 159.9 (C-2, C-8) , 144.8 (C-4, C-6) , 115.0 (C-3,
3
Bim
Glc
Glc
Glc
Glc
Glc
C-7) , 81.8 (C-1,1’) , 76.5 (C-5,5’) , 73.3 (C-3,3’) , 69.5 (C-2,2’) , 67.9 (C-4,4’) ,
Glc
Bim
Glc
Bim
6
1.8 (C-6,6’) , 22.3 (CH ) , 20.7, 20.6 (COCH ) , 7.2 (CH ) . HRMS m/z calc. for
2 3 3
+
C H N O S [M+H] : 917.231 Found: 917.230.
38
48
2
20 2
4
.18. syn-(CH S-β-GalAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
galactopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
8b)
(
According to general method A, the reaction of 1a (200 mg, 0.57 mmol) with 4b (438
mg, 1.2 mmol) and triethylamine (0.2 mL, 1.4 mmol) gave 8b (401 mg, 77 %) as a pale
27
D
1
yellow solid. R 0.61 (4:1 toluene-isopropanol), [α] – 86 (c 0.25 CDCl ). H NMR (CDCl ,
f
3
3
Gal
Gal
5
00 MHz): δ (ppm) 5.45 (d, 2H, H-4,4’ , J = 1.5 Hz), 5.20 (t, 2H, H-2,2’ , J = J2,3 =
3,4 1,2
Gal
Gal
Gal
1
0.0 Hz), 5.07 (dd, 2H, H-3,3’ ), 4.52 (d, 2H, H-1,1’ ), 4.10 (m, 8H, H-6a,6a’ , H-
2
4

ACCEPTED MANUSCRIPT
Gal
Bim
Bim
Gal
6
2
1
b,6b’ , CH2
, CH2b,b’ ), 3.96 (t, 2H, H-5,5’ , J = J5,6b = 6.5 Hz), 2.20, 2.08, 2.07,
a,a’
5,6a
Gal
Bim 13
.00 (s, 24H, 8xCOCH3 ), 1.96 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm)
3
Gal
Bim
Bim
Bim
70.2, 169.6 (COCH ) , 159.8 (C-2, C-8) , 144.8 (C-4, C-6) , 114.6 (C-3, C-7) , 82.7
3
Gal
Gal
Gal
Gal
Gal
(
C-1,1’) , 75.0 (C-5,5’) , 71.2 (C-3,3’) , 67.0 (C-2,2’) , 66.6 (C-4,4’) , 61.3 (C-
Gal
Bim
Gal
Bim
6
,6’) , 22.6 (CH ) , 20.5 (COCH ) , 6.9 (CH ) . HRMS m/z calc. for C H O N S
2 3 3 38 48 20 2 2
+
[
M+H] : 917.231 Found: 917.232.
4
.19. syn-(CH S-β-ManAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 4 3
mannopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
8βc)
(
According to general method A, the reaction of 1a (200 mg, 0.57 mmol) with 4βc (438
mg, 1.2 mmol) and triethylamine (0.2 mL, 1.4 mmol) gave 8βc (382 mg, 73 %) as a pale
27
D
1
yellow solid. R 0.61 (4:1 toluene-isopropanol), [α] – 106 (c 0.25 CDCl ). H NMR (CDCl ,
f
3
3
Man
Man
5
1
6
=
00 MHz): δ (ppm) 5.50 (d, 2H, H-2,2’ , J2,3 = 3.0 Hz), 5.26 (t, 2H, H-4,4’ , J3,4 = J =
4,5
Man
Man
Man
0.0 Hz), 5.11 (dd, 2H, H-3,3’ ), 5.01 (s, 2H, H-1,1’ ), 4.26 (dd, 2H, H-6a,6a’ , J5,6a
=
Man
Bim
Bim
.0 Hz, J6a,6b = 12.5 Hz), 4.17 (m, 4H, H-6b,6b’ , CH2a,a’ ), 3.93 (d, 2H, CH2b,b’ , JCH2a,b
Man
Man
15.0 Hz), 3.88 (m, 2H, H-5,5’ ), 2.18, 2.10, 2.02, 1.92 (s, 24H, 8xCOCH3 ), 1.88 (s,
Bim 13
Man
6
1
5
2
9
H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.0, 169.7, 169.3 (COCH )
,
3
3
Bim
Bim
Bim
Man
58.7 (C-2, C-8) , 142.6 (C-4, C-6) , 114.9 (C-3, C-7) , 78.7 (C-1,1’) , 75.8 (C-
Man
Man
Man
Man
Man
Bim
,5’) , 71.0 (C-3,3’) , 69.8 (C-2,2’) , 65.2 (C-4,4’) , 62.5 (C-6,6’) , 22.8 (CH2)
,
Man
Bim
+
0.2 (COCH3) , 6.5 (CH3) . HRMS m/z calc. for C H N O S [M+H] : 917.231 Found:
38
48
2
20 2
17.233.
2
5

ACCEPTED MANUSCRIPT
4
.20. syn-(CH S-α-ManAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-α-D-
2
4
3
mannopyranosyl)thio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
8αc)
(
According to general method A, the reaction of 1a (176 mg, 0.50 mmol) with 4αc (376
mg, 1.0 mmol) and triethylamine (0.2 mL, 1.4 mmol) furnished 8αc (97 mg, 21 %) as a pale
27
D
1
yellow solid. R 0.61 (4:1 toluene-isopropanol), [α] + 4 (c 0.25 CDCl ). H NMR (CDCl ,
f
3
3
Man
5
4
00 MHz): δ (ppm) 5.34 (dd, 2H, H-2,2’ , J1,2 = 1.5 Hz, J2,3 = 3.5 Hz), 5.30 (t, 2H, H-
Man
Man
Man
,4’ , J3,4 = J = 10.0 Hz), 5.18 (dd, 2H, H-3,3’ ), 5.12 (d, 2H, H-1,1’ ), 4.34 (m, 2H,
4,5
Man
Man
H-5,5’ ), 4.27 (dd, 2H, H-6a,6a’ , J5,6a = 6.0 Hz, J6a,6b = 12.5 Hz), 4.21 (dd, 2H, H-
Man
Bim
Bim
6
2
b,6b’ , J5,6b = 2.0 Hz), 4.00 (d, 2H, CH2a,a’ , JCH2a,b = 15.0 Hz), 3.91 (d, 2H, CH2b,b’ ),
Man
Bim 13
.17, 2.13, 2.08, 1.99 (s, 24H, 8xCOCH3 ), 1.92 (s, 6H, 2xCH3 ), C NMR (CDCl , 125
3
Man
Bim
Bim
MHz): δ (ppm) 169.9, 169.3, 169.0 (COCH3) , 159.0 (C-2, C-8) , 142.9 (C-4, C-6)
,
Bim
Man
Man
Man
Man
1
15.6 (C-3, C-7) , 80.0 (C-1,1’) , 69.6 (C-3,3’) , 69.1 (C-5,5’) , 69.0 (C-2,2’)
,
Man
Man
Bim
Man
Bim
6
5.6 (C-4,4’) , 62.1 (C-6,6’) , 22.6 (CH2) , 20.4, 20.2 (COCH3) , 6.6 (CH3)
.
+
HRMS m/z calc. for C H N O S [M+Na] : 939.214 Found: 939.216.
3
8
48
2
20 2
4
.21. syn-(CH S-β-LacAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-
D
-galactosyl-
2
7
3
(1→4)-2,3,6-tri-O-acetyl-1-thio-β-D-glucosyl)thio]methyl}-3,7-dimethyl-1,5-
diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione (8d)
According to general method A, the reaction of 1a (100 mg, 0.29 mmol) with 4d (392
mg, 0.60 mmol) and triethylamine (0.1 mL, 0.72 mmol) furnished 8d (383 mg, 90 %) as a
27
D
1
pale yellow solid. R 0.41 (4:1 toluene-isopropanol), [α] – 81 (c 0.25 CDCl ). H NMR
f
3
Gal
Glc
(
CDCl , 500 MHz): δ (ppm) 5.27 (d, 2H, H-4,4’ , J = 5.0 Hz), 5.13 (t, 2H, H-3,3’ , J
=
3
3,4
2,3
Gal
Gal
J = 10.0 Hz), 5.01 (t, 2H, H-2,2’ , J = J = 10.0 Hz), 4.89 (dd, 2H, H-3,3’ ), 4.84 (t,
3
,4
1,2
2,3
Glc
Gal
Glc
Glc
2
H, H-2,2’ ), 4.43 (m, 4H, H-1,1’ , H-1,1’ ), 4.38 (d, 2H, H-6a,6a’ , J
= 10.0 Hz),
a,6b
6
2
6

ACCEPTED MANUSCRIPT
Bim
Glc
Gal
Gal
Glc
3
1
1
.98 (m, 10H, 2xCH2 , H-6b,6b’ , H-6a,6a’ , H-6b,6b’ ), 3.83 (t, 2H, H-4,4’ , J
=
4,5
Gal
Glc
0.0 Hz), 3.69 (t, 2H, H-5,5’ , J5 = J5,6b = 9.5 Hz), 3.55 (m, 2H, H-5,5’ ), 2.07, 2.05,
,6a
Lac
Bim 13
.98, 1.97, 1.96, 1.88, 1.83 (s, 42H, 14xCOCH3 , 2xCH3 ), C NMR (CDCl , 125 MHz):
3
Lac
Bim
Bim
δ (ppm) 170.4, 169.9, 169.4, 169.0 (COCH ) , 159.7 (C-2, C-8) , 144.6 (C-4, C-6)
,
3
Bim
Gal
Glc
Glc
Glc
1
14.7 (C-3, C-7) , 100.9 (C-1,1’) , 81.7 (C-1,1’) , 77.4 (C-5,5’) , 75.6 (C-4,4’) , 73.1
Glc
Gal
Gal
Glc
Gal
(
C-3,3’) , 70.8 (C-5,5’) , 70.6 (C-3,3’) , 69.8 (C-2,2’) , 69.0 (C-2,2’) , 66.5 (C-
Gal
Glc
Gal
Bim
Lac
Bim
4
,4’) , 61.9 (C-6,6’) , 60.7 (C-6,6’) , 22.4 (CH ) , 20.5, 20.4 (COCH ) , 7.0 (CH )
.
2
3
3
+
HRMS m/z calc. for C H N O S [M+H] : 1493.400 Found: 1493.404.
6
2
80
2
36 2
4
.22. General method B for the preparation of disulfido-glycosides
Compound 1d was dissolved in DMF with stirring under nitrogen atmosphere at room
temperature and the appropriate 9(a-c) added. Stirring was continued at room temperature for
0 min then the reaction mixture was poured into water. The solids deposited were filtered,
rinsed with water and dried. The crude products were purified by column chromatography
9.5:0.5 toluene:isopropanol) to give 10(a-c) disulfido-glycosides.
3
(
4
.23. syn-(CH S -β-GlcAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 2 4 3
glucopyranosyl)dithio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione
10a)
(
According to general method B, the reaction of 1d (130 mg, 0.51 mmol) with 9a (650
mg, 1.3 mmol) gave 10a (406 mg, 81 %) as a pale yellow solid. R 0.58 (4:1 toluene-
f
27
1
isopropanol), [α]D – 161 (c 0.25 CDCl ). H NMR (CDCl , 500 MHz): δ (ppm) 5.38 (t, 2H,
3
3
Glc
Glc
Glc
H-2,2’ , J = J = 9.5 Hz), 5.29 (t, 2H, H-3,3’ ), 5.15 (t, 2H, H-4,4’ , J = J = 9.5
1
,2
2,3
3,4
4,5
Glc
Bim
Glc
Hz), 4.59 (d, 2H, H-1,1’ ), 4.27 (m, 6H, 2xCH , H-6a,6a’ , J
= 12.5 Hz, J5,6a = 5.0
2
6a,6b
Glc
Glc
Hz), 3.96 (d, 2H, H-6b,6b’ ), 3.85 (m, 2H, H-5,5’ ), 2.10, 2.06, 2.03 (s, 24H,
2
7

ACCEPTED MANUSCRIPT
Glc
Bim 13
8
1
xCOCH3 ), 1.97 (s, 6H, 2xCH3 ), C NMR (CDCl , 125 MHz): δ (ppm) 170.5, 169.9,
3
Glc
Bim
Bim
Bim
69.3, 169.0 (COCH ) , 159.9 (C-2, C-8) , 144.5 (C-4, C-6) , 116.1 (C-3, C-7) , 86.1
3
Glc
Glc
Glc
Glc
Glc
(
C-1,1’) , 77.3 (C-5,5’) , 73.4 (C-3,3’) , 68.6 (C-2,2’) , 67.8 (C-4,4’) , 61.8 (C-
Glc
Bim
Glc
Bim
6
,6’) , 32.7 (CH ) , 20.6, 20.5 (COCH ) , 7.4 (CH ) . HRMS m/z calc. for
2 3 3
+
C H N O S [M+Na] : 1003.158 Found: 1003.153.
38
48
2
20 4
4
.24. syn-(CH S -β-GalAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-D-
2 2 4 3
galactopyranosyl)dithio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-
dione (10b)
According to general method B, the reaction of 1d (68 mg, 0.27 mmol) with 9b (340
mg, 0.67 mmol) gave 10b (185 mg, 71 %) as a pale yellow solid. R 0.58 (4:1 toluene-
f
27
1
isopropanol), [α]D – 108 (c 0.25 CDCl ). H NMR (CDCl , 500 MHz): δ (ppm) 5.55 (t, 2H,
3
3
Gal
Gal
H-2,2’ , J = J = 10.0 Hz), 5.49 (dd, 2H, H-4,4’ , J = 3.0 Hz, J4,5 = 0.5 Hz), 5.12 (dd,
1
,2
2,3
3,4
Gal
Gal
Gal
Gal
Bim
a,a’
2
H, H-3,3’ ), 5.11 (d, 2H, H-1,1’ ), 4.19 (m, 6H, H-6a,6a’ , H-6b,6b’ , CH
), 4.06
2
Gal
Bim
(
td, 2H, H-5,5’ , J5 = J5,6b = 6.5 Hz, J4,5 = 0.5 Hz), 4.01 (d, 2H, CH2a,a’ , JCH2a,b = 14.5
,6a
Gal
Bim 13
Hz), 2.22, 2.07, 2.05, 2.01 (s, 24H, 8xCOCH3 ), 1.97 (s, 6H, 2xCH3 ), C NMR (CDCl ,
3
Gal
Bim
Bim
1
25 MHz): δ (ppm) 170.4, 170.0, 169.4 (COCH ) , 160.0 (C-2, C-8) , 144.3 (C-4, C-6)
,
3
Bim
Gal
Gal
Gal
Gal
1
16.2 (C-3, C-7) , 87.0 (C-1,1’) , 75.5 (C-5,5’) , 71.5 (C-3,3’) , 67.2 (C-2,2’) , 66.1
Gal
Gal
Bim
Gal
Bim
(
C-4,4’) , 61.9 (C-6,6’) , 32.7 (CH ) , 20.7, 20.6 (COCH ) , 7.5 (CH ) . HRMS m/z
2
3
3
+
calc. for C H N O S [M+Na] : 1003.158 Found: 1003.152.
3
8
48
2
20 4
4
.25. syn-(CH S -β-ManAc ,CH )B, 4,6-bis-{1-[(2,3,4,6-tetra-O-acetyl-β-
D
-
2
2
4
3
mannopyranosyl)dithio]methyl}-3,7-dimethyl-1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-
dione (10c)
2
8