Multivalent glycoclusters constructed by chiral self-assembly of mannose functionalized perylene bisimide

Article abstract of DOI:10.1039/c2ob27052h

Water-soluble perylene bisimide derivative 7 modified with six mannoses was synthesized and its self-assembled properties were studied by UV-Vis and CD spectroscopy, which revealed an interesting self-assembly with a solvent-tuning chiral conformation in H₂O-DMSO solution. As H₂O was added to the DMSO solution until a 60% (or 70%) v/v proportion was achieved, the self-assembly of the mannose functionalized compound 7 exhibited a left-handed helical conformation. More interestingly, when the volume of H₂O constituted beyond 85% of the solution, the conformation of the self-assembly turned out to be a right-handed helical conformation. Furthermore, the binding interactions between the self-assembly of compound 7 and Con A were investigated by turbidity assay, CD spectra, TEM and SEM images, and ELLA experiment, which indicated that the self-assembly of compound 7 as multivalent glycoclusters exhibited specific binding to Con A with an IC₅₀ value of 24 μM (144 μM, valency corrected), 10 times stronger than the reference compound (α-MMP).

Full text of DOI:10.1039/c2ob27052h

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Multivalent
Glycoclusters
Constructed
by
Chiral
Self-assembly of Mannose Functionalized Perylene Bisimide
Ke-Rang Wang,*a,b Hong-Wei An,a,b Yue-Qing Wang,a,b Jin-Chao Zhang,a,b and Xiao-Liu
Li*a,b
a
Key Laboratory of Chemical Biology of Hebei Province, College of Chemistry and
Environmental Science, Hebei University, Baoding 071002, China
b
Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of
Education, Hebei University, Baoding 071002, China
Tel & Fax: (+86)312-5971-116; E-mail: kerangwang@hbu.edu.cn; lixl@hbu.cn

Page 2 of 7
Organic BiomoleculOarrganic & Biomolecular Chemistry
Chemistry
Dynamic Article Links ►
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Cite this: DOI: 10.1039/c0xx00000x
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ARTICLE TYPE
Multivalent Glycoclusters Constructed by Chiral Self-assembly of
Mannose Functionalized Perylene Bisimide
Ke-Rang Wang,*a,b Hong-Wei An,a,b Yue-Qing Wang,a,b Jin-Chao Zhang,a,b and Xiao-Liu Li*a,b
Received (in XXX, XXX) Xth XXXXXXXXX 20XX, Accepted Xth XXXXXXXXX 20XX
5
DOI: 10.1039/b000000x
A waterꢀsoluble perylene bisimide derivative 7 modified with six mannoses was synthesized and its selfꢀ
assembled properties were studied by UVꢀVis and CD spectra, which revealed an interesting selfꢀ
assembly with solventꢀtuning chiral conformation in H2O/DMSO solution. As H2O was added to the
DMSO solution up to 60 % (or 70 %), the selfꢀassembly of the mannose functionalized compound 7
10 exhibited a leftꢀhanded helical conformation. More interestingly, when the volume of H2O beyond 85%,
the conformation of the selfꢀassembly turned to be a rightꢀhanded helical conformation. Furthermore, the
binding interactions between the selfꢀassembly of compound 7 and Con A were investigated by turbidity
assay, CD spectra, TEM and SEM images, and ELLA experiment, which indicated that the selfꢀassembly
of compound 7 as multivalent glycoclusters exhibited specific binding to Con A with the IC50 value of 24
15 ꢁM (144 ꢁM, valency corrected), 10 times stronger than the reference compound (αꢀMMP).
30 the number of the studies on supramolecular glycoclusters based
on πꢂꢂꢂπ stacking interactions was limited.6
Introduction
Carbohydrateꢀprotein (lectin) interactions have been recognized
to play a very important function in a wide variety of biological
processes, such as cell growth regulation, differentiation,
20 adhesion, cancer cell metastasis, cellular trafficking,
inflammation by bacteria and viruses, and the immune response.1
Because of their essential role in the maintenance of the diverse
biological systems, much attention is currently focused on the
design and synthesis of carbohydrateꢀbased units for investigating
25 the carbohydrateꢀprotein (lectin) interaction events.2 However,
due to the relatively low affinity of monosaccharideꢀprotein
(lectin) interactions,3 synthetic glycoclusters as multivalent model
systems have been employed to enhance the binding efficiency.4
Though multivalent glycoclusters have been widely investigated,5
Scheme 1 Structural illustration of compound 7, the possible selfꢀ
35 assembly mode and a schematic representation of the possible mode of
interaction with Con A.
a Key Laboratory of Chemical Biology of Hebei Province, College of
Chemistry and Environmental Science, Hebei University, Baoding
071002, China
b
Brunsveld and coworkers6a designed and synthesized a
mannose functionalized discꢀshaped compound which selfꢀ
assembled to polyvalent columnar supramolecular polymers, and
40 found that selfꢀassembled polymers exhibited binding interactions
with Concanavalin A (Con A) and Escherichia coli. Lee and
coworkers6bꢀd constructed supramolecular multivalent ligands by
mannose modified rodꢀcoil molecules which showed selectively
binding with Con A and Escherichia coli strain ORN178. Khiar
45 and Ellervik reported that supramolecular selfꢀassemblies of
carbohydrate modified polydiacetylene derivatives exhibited
Key Laboratory of Medicinal Chemistry and Molecular Diagnosis
of Ministry of Education, Hebei University, Baoding 071002, China
Tel & Fax: (+86)312ꢀ5971ꢀ116; Eꢀmail: kerangwang@hbu.edu.cn;
lixl@hbu.cn
† Electronic Supplementary Information (ESI) available: NMR
spectra and HRMS of compound 2, 4, 6 and 7; NMR spectra, FTꢀIR
spectra and HRMS of compound 6 and 7; Concentrationꢀdependent
and temperatureꢀdependent UV−Vis spectra of 7; CD spectra of
compound 7 upon addition of Con A in PBS buffer; TEM image of
the selfꢀassembly of compound 7 in water; Inhibition curve of αꢀ
MMP. See DOI: 10.1039/b000000x/
This journal is © The Royal Society of Chemistry [year]
[journal], [year], [vol], 00–00 | 1

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colorimetric detection of Con A 7 and efficient inhibitors of 40 (Fig. S1ꢀS14).
Adenoviridae binding (IC50
=
0.9 ꢃM) and Adenoviridae
UVꢀVis spectra of compound 7 in different volume ratios of
DMSO/H2O (Fig. 1) showed that 7 existed in the typically
nonaggregated state in DMSO solution. 14 Three distinguishable
absorption bands between 450 and 650 nm are observed as 588
infectivity (IC50 = 0.7 ꢃM).8 Brunsveld and Schenning et al9
reported multiꢀtargeted fluorescent nanoparticles based on the
selfꢀassembly of πꢀconjugated amphiphilic oligomers, which
5
were capable of selective binding to Con A and Escherichia coli 45 nm, 546 nm and 452 nm, and the most intensity appears at the
strain ORN178. Recently, we reported a chiral waterꢀsoluble selfꢀ
assembly constructed by Dꢀlactose functionalized perylene
bisimide derivative which as multivalent glycoclusters exhibited
first band (588 nm). These characteristic properties were typical
for the modified perylene bisimide derivative at bay areas. 15
Upon addition of water, the absorption of 7 changed obviously.
The intensities of the bands of 588 nm, 546 nm and 452 nm
10 specific binding with the lectin of peanut agglutinin (PNA).10
Herein, we report the synthesis and chiral aggregation 50 decreased, and the absorption bands of 588 nm and 546 nm were
behavior of a very waterꢀsoluble perylene bisimide derivative
bearing Dꢀmannose moiety and its binding interactions with Con
A (Scheme 1). Perylene bisimide moiety was employed as a
15 building block to construct multivalent ligands because of their
finally bathochromic shift to 598 nm and 556 nm, respectively, as
a result of π⋅⋅⋅π aggregation,15 indicating the pronounced π⋅⋅⋅π
aggregation of 7 in water.
Concentrationꢀdependent UVꢀVis spectra of compound 7
strong π⋅⋅⋅π stacking interactions between backbones, which have 55 (Fig. S15) showed that compound 7 was in the aggregate state
been widely used to construct nanoscopic architectures.11 The
mannose moiety was not only as waterꢀsoluble and chiral group
but also as binding site for Con A.5
even at a low concentration of 3.0 × 10–6 M, where the maximal
absorption band appears at 551 nm. Upon increasing the
concentration, the apparent absorption coefficients (ε) of the
absorption bands slightly decreased, indicating strong π⋅⋅⋅π
20 Results and Discussion
60 stacking interaction between the backbones of compound 7 in
16
water.
Temperatureꢀdependent UVꢀVis spectra (Fig. S16)
showed that the aggregate of 7 was difficult to deaggregate by
controlling the temperature, which also implies the much stronger
π⋅⋅⋅π stacking interaction of compound 7 in water.
50k
DMSO
10% H2O
20% H2O
40k
30% H2O
40% H2O
50% H2O
30k
60% H2O
70% H2O
80% H2O
20k
ε
85% H2O
90% H2O
95% H2O
10k
H2O
0
400
500
600
700
Wavelength/nm
65
Fig. 1 UVꢀVis spectra of compound 7 (4 × 10–5 M) in different volume
ratios of DMSO/H2O.
Scheme 2 Reagents and conditions: a) isoꢀpropyl alcohol, reflux; b)
K2CO3, NMP, reflux; c) Sodium ascorbate, CuSO4, THF/H2O, 5 h; d)
25 NaOH, MeOH/H2O.
The distinguishable selfꢀassembly behaviors of 7 in different
volume ratios of DMSO/H2O were identified by circular
70 dichroism (CD) spectroscopy. As shown in Fig. 2a, no CD signal
was observed for freely soluble 7 in DMSO solution. However,
the CD spectra changed as water was added to the solution. As 60
% H2O was added to the DMSO solution, a bisignate CD
spectrum with a positive CD signal at ca. 387 nm and two
75 negative CD signals at ca. 587 nm and 629 nm were observed.
With the further addition of H2O (to 70%), the intensity of the
CD signal increased. The bisignate positive/negative signal with
increasing wavelength indicates that compound 7 adopted a leftꢀ
handed helical conformation. 17 Much more interestingly, with the
80 gradual increase of H2O to 80% in volume, the CD spectrum
changed with a negative CD signal at ca. 400 nm, a positive CD
signal at ca. 520 nm and a negative CD at ca. 578 nm along
increasing wavelength, implying that the selfꢀassembly of
compound 7 existed a chiral comformation transfering state from
The αꢀDꢀmannoseꢀmodified perylene bisimide 7 was synthesized
through four steps as shown in Scheme 2. Firstly, the
tetrachloroperylene tetracarboxylic acid dianhydride
1 was
converted to the corresponding propargyl bisimide using
30 propargyl amine. Subsequent nucleophilic substitution of the four
chlorine atoms by phenolate 312 gave the compound 4 with six
propargyl groups. The target compound 7 was synthesized by the
reaction
of
2ꢀazidoethylꢀ2,3,4,6ꢀtetraꢀOꢀacetylꢀαꢀDꢀ
mannopyranoside (5)13 with 4 under standard Cu (I) conditions in
35 the presence of substoichiometric amount of CuSO4 and sodium
ascorbate in a homogeneous THF/water solution, which afforded
fully substituted perylene bisimide derivative 6, and then
deprotection of the acetyl groups. These compounds were fully
characterized by NMR spectroscopy, FTꢀIR spectra and HRMS
2
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leftꢀhanded helical conformation to lightꢀhanded helical
conformation. When H2O was increased up to 85%, the selfꢀ
assembly of compound 7 exhibited the opposite behavior in terms 35 formation of crossꢀlinked complexes in real time.
model because of the selective binding of Con A to mannose
residue. 18 Turbidity assay has been readily used to monitor the
19
Upon
of chirality with two negative CD signals at ca. 376 nm and 399
nm and two positive CD signals at ca. 542 nm and 627 nm. With
further increasing the volume of H2O, the intensity of the CD
signal gradually increased. The bisignate CD signal is a classic
addition of the selfꢀassembly of 7 to the solution of Con A in PBS
5
buffer (pH = 7.2, 10 mM) containing 0.1 mM MnCl2 and 0.1 mM
CaCl2, the turbidity immediately increased, which indicated the
possible carbohydrateꢀprotein binding interaction. In order to
signature for excitonic coupling as
a result of chiral 40 demonstrate that the turbidity increase was resulted from the
aggregation.17 The shape of the bisignate negative/positive Cotton
carbohydrateꢀprotein interactions, a large excess of αꢀmethylꢀDꢀ
mannopyranoside (αꢀMMP) was added to inhibit glycoclusters–
Con A binding. Upon addition of αꢀMMP, substantial decrease in
turbidity was observed (Fig. 3). The disappearance of turbidity
10 effect with increasing wavelength indicated that compound 7
adopted a rightꢀhanded supramolecular stacks induced by the
chiral Dꢀmannose moieties in water. It is well known that the
helical conformations of the πꢀconjugated molecules with chiral 45 clearly indicated that the observed increase in optical density was
side chains afford a preferential helicity on the higherꢀorder
15 supramolecular assemblies. 17
a result of carbohydrate–protein interactions.20 In addition, as the
control experiment, the turbidity of the mixture of 7 with PNA
and bovine serum albumin (BSA) exhibited no obvious change,
indicative of no binding occurred between the selfꢀassembly of 7
50 and PNA or BSA. The results showed that the selfꢀassembly of
compound 7 as multivalent glycoclusters exhibited specifically
and selectively binding properties with Con A. Moreover, the CD
spectra (Fig. S17) were used to investigate the interactions of the
selfꢀassembly of compound 7 with Con A. Upon addition of
55 different concentrations of Con A, the CD signal of the selfꢀ
assembly of 7 showed slightly change, indicating the rightꢀ
handed supramolecular selfꢀassembly of 7 had no change when
binding with Con A.
a)
12
8
4
0
DMSO
ε
ꢀ
60 % H2O
70 % H2O
-4
80 % H2O
85 % H2O
90 % H2O
95 % H2O
-8
-12
H2O
400
500
600
700
Con A
BSA
PNA
0.20
Wavelength/nm
0.16
20
15
10
5
b
)
0.12
0.08
0.04
0.00
0
10−6
10−6
10−5
10−5
10−4
M
M
M
M
M
ε
ꢀ
3
6
1
5
1
2
×
×
×
×
×
×
-5
0
500
1000
1500
2000
2500
-10
-15
-20
Time (s)
60 Fig. 3 Turbidity assay of 7 (2.0 × 10−5 M) at 750 nm with Con A (1 mg,
black line), BSA (1 mg, red line) and PNA (1 mg, blue line) in PBS
buffer (pH = 7.2, 10 mM, 0.1 mM MnCl2 and 0.1 mM CaCl2), and αꢀ
MMP (1.0 × 10−2 M) was added to the mixture of Con A with 7 as an
inhibitor at 1300 s.
10−4M
400
500
600
700
Wavelength/nm
Fig. 2 a) CD spectra of compound 7 (4 × 10–5 M) in different volume
ratios of DMSO/H2O; b) concentration dependent CD spectra of 7 from 3
20 × 10–6 M to 2 × 10–4 M in water.
65
In addition, the TEM and SEM images were used to
characterize the interactions of the selfꢀassembly of 7 with Con A
lectin. In water, the nanofibers of the selfꢀassembly of compound
7 was observed (Fig. S18). After mixing of the selfꢀassembly of 7
with Con A for 2 h, the TEM image (Fig. 4a) of the mixture
70 showed obvious pellets with the diameters of ca. 10 nm along the
nanofibers. The SEM image (Fig. 4b) was also used to investigate
the morphological properties of the interaction of the selfꢀ
assembly of 7 with Con A. It was obviously found that after
immersion in Con A solution for 2 h the surface of the nanofibers
75 became much more rough with obvious nanoꢀballs because of the
aggregation of Con A.
Moreover, the concentrationꢀdependent CD spectra of 7
were also investigated (Fig. 2b). The intensities of the positive
and negative Cotton effects in CD signal of 7 showed nearly twoꢀ
fold increase as the concentration increased from 3 × 10–6 M to 2
25 × 10– M, which furtherly supported the formation of the
supramolecular chiral selfꢀassembly of 7 in water solution
because of π⋅⋅⋅π aggregation.
The formation of the supramolecular chiral selfꢀassembly of
compound 7 coated by mannoses suggested that they would be
30 used as supramolecular glycoclusters. To investigate the specific
lectinꢀcarbohydrate interactions between the supramolecular
glycoclusters, turbidity assay was carried out with a Con A lectin
4
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Conclusions
A waterꢀsoluble perylene bisimide derivative modified with six
mannoses was synthesized and fully characterized. Benefiting
25 from the modification of Dꢀmannose moiety, compound 7
exhibited supramolecular stacks in water with chiral
characteristic induced by the chiral Dꢀmannose moiety.
Interestingly, the selfꢀassembly of compound 7 exhibited leftꢀ
handed helical conformation when 60 % (or 70 %) H2O was
30 added to the DMSO solution. Much more differently, continued
increase in H2O volume beyond 85%, the selfꢀassembly showed
rightꢀhanded helical conformation. Furthermore, the selectively
binding interactions between the selfꢀassembly of compound 7
with Con A were investigated by turbidity assay and CD spectra,
35 which showed that the selfꢀassembly as multivalent glycoclusters
exhibited specifically and selectively binding properties with Con
A. The morphology of the selfꢀassembly with Con A was
investigated by TEM and SEM images, obvious pellets along the
nanofibers were found. ELLA experiment indicated that the selfꢀ
40 assembly of 7 as multivalent glycoclusters exhibited strong
binding with Con A with the IC50 value of 24 ꢁM (144 ꢁM,
valency corrected), 10 times stronger than the reference
compound (αꢀMMP).
Experimental section
Fig. 4 TEM (a) and SEM (b) imges of the complexes between the selfꢀ 45 Materials: 1H NMR and 13C NMR spectra were recorded on a
assembly of compound 7 with Con A.
Bruker 600 Spectrometer. HRMS was performed on an IonSpec
QFTꢀMALDI MS (Varian). UV−Vis spectra were recorded in a
To quantify the multivalent binding of the selfꢀassembly of 7
5
quartz cell (light path 10 mm or 5 mm) on a Shimadzu UVꢀ3600
with lectin, an enzyme linked lectin assay (ELLA) was
spectrophotometer equipped with
a
Sꢀ1700 temperature
performed. The experiments using horseradish peroxidase labeled
Con A (HRPꢀCon A) as the lectin and yeast mannan as the
surface fixedꢀligand were carried out in 96ꢀwell plates. After
50 controller. Turbidity assay of the ligandꢀprotein binding was
mensurated on a Shimadzu UVꢀ3600 spectrophotometer with
quartz microꢀcuvette (100 ꢁL volume, 1 cm path length).
Transmission Electron Microscope (TEM) image was recorded
on JEMꢀ2100 (JEOL). Scanning electron microscopic (SEM)
55 image was recorded on JSMꢀ7500F (JEOL). Circular dichroism
spectra were performed on MOSꢀ450 (BioLogic).
Turbidity assay: Turbidity assays were performed by adding
100 ꢃL of the Con A solution (1 mg/mL) to a dry quartz microꢀ
cuvette (100 ꢃL volume, 1 cm path length). A solution of the
60 compound 7 in PBS buffer was quickly added (10 ꢃL at 2.0 ×
10−5 M) and mixed vigorously. Absorbance data were recorded at
750 nm for 1300 s. The inhibition assay was performed by adding
the αꢀMMP (1.0 × 10−2 M) solution to the mixture of Con A with
7 at 1300 s.
65 Enzyme-linked lectin assays (ELLA): 96ꢀwell microtiter plates
(Nunc Maxisorb) were coated with yeast mannan (10 ꢃg/mL in
10 mM PBS, pH = 7.2 with 0.1 mM Mn2+ and Ca2+) overnight at
room temperature. Then each well was washed three times with
250 ꢃL PBST buffer (PBS with 0.05 % (v/v) Tween 20). The
70 wells were then blocked for 1 h at 37 °C using PBSA (1% BSA in
PBS, 100 ꢃL/well). Washing was repeated as described above.
Separately, serial dilutions of inhibitors (60 ꢃL/well) were
performed in nontreated mixing plates. Stock solutions of
compound 7 or αꢀMMP in PBS buffer were then used. These
75 wells were completed with a solution of 60 ꢃL/well (0.01mg/mL)
Horseradish Peroxidase (HRP) labeled Concanavalin A and the
plates were incubated 1 h at 37 °C. These solutions were
10 preincubation with different concentrations of 7, and αꢀMMP as
the reference compound, the binding of HRPꢀCon A to mannan
was measured by spectroscopic method. The IC50 value for αꢀ
MMP was around 1400 ꢁM (Fig. S19) in our assay, which is
similar with the reported result.21 By contrast, the IC50 value for
15 the selfꢀassembly of compound 7 is around 24 ꢁM (144 ꢁM,
valency corrected, Fig. 5). The relative valencyꢀcorrected binding
of 7 was 10 times stronger than the reference compound,
remarkably exhibiting the multivalent inhibition effect of the selfꢀ
assembly.
80
70
60
50
40
30
20
10
0
0
200
Inhibitor concentration (
Fig. 5 Inhibition curve of compound 7 for determination of the IC50.
400
600
800
1000
ꢁM)
20
4
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transferred into the yeast mannan coated plates, incubated 1 h at
found 3577.0631.
37 °C. The washing step was repeated and 50 ꢃL/well ABTS 60 Synthesis of compound 7: Compound 6 (711 mg, 0.2 mM) was
(0.25 mg/ml citrate buffer 0.2 M, pH = 4.0 with 0.015 % (v/v)
H2O2) were added. Color development was stopped after 20 min
by adding 50 ꢃL/well 1M H2SO4. Light absorption was measured
at 415 nm.
dissolved in MeOH (20 mL), a solution of sodium hydroxide (2.0
mL, 8 mM in H2O) was added, and the reaction mixture was
stirred at room temperature until disappearance of the starting
material. The mixture was neutralized with dilute hydrochloric
65 acid (2 M). And the solid was filtered, washed with MeOH. Then
the solid was dissolved in water, the solution was put in a
cellulose dialysis tube (cutoff 1000), dialyzed against water for 2
d, and lyophilized to give the desired solid polymer. 7 (458 mg)
was obtained with yield of 90 %. m. p. 143.8ꢀ145.4 ºC; FT–IR
5
% Inhibition = (A(no inhibitor) ꢀ A(with inhibitor) )/A(no inhibitor) ×100
Synthesis of compound 2: Compound 1 (2.0 g, 3.77 mM) and
propargylamine (0.72 mL, 11.3 mM) were mixed in isoꢀpropyl
10 alcohol (100 mL). The reaction mixture was refluxing under N2
for 24 h. After cooling to room temperature, the reaction mixture
was poured into ice water, the solid compound was filtered,
washed by water. The residue was purified by silicaꢀgel column
chromatography using CH2Cl2/petroleum ether (v/v = 1.5/1) as
15 the eluent to give the product as a red powder (1.2 g) at yield of
52.8 %. m. p. > 250°C. 1H NMR (600 MHz, CDCl3, ppm): δ 2.24
(t, J = 1.8 Hz, 2H, ꢀC≡CH), 4.99 (s, 4H, ꢀCH2ꢀ), 8.73 (s, 4H,
peryleneꢀH); 13C NMR (150 MHz, CDCl3, ppm): δ 29.6, 71.2,
122.9, 123.3, 128.8, 133.2, 135.5, 161.4. HRMS: calcd for
20 C30H10Cl4N2O4, 603.9365; found 603.9371.
1
70 (KBr) v [cm–1]: 1697, 1658, 1589, 1500; H NMR (600 MHz,
DMSOꢀd6, ppm): δ 3.17 (m, 6H), 3.34ꢀ3.42 (m, 12H), 3.56 (m,
6H), 3.63 (d, J = 10.8 Hz, 4H), 3.75 (m, 2H), 3.82 (m, 4H), 3.89
(m, 2H), 3.97 (s, 4H), 4.44ꢀ4.81 (m, 25H), 5.13 (s, 8H), 5.23 (s,
4H), 7.02ꢀ7.08 (m, 16H, ArꢀH), 7.86 (s, 4H, Triaz1ꢀH); 7.99 (s,
75 2H, Triaz2ꢀH); 8.22 (s, 4H, peryleneꢀH); 13C NMR (100 MHz,
DMSOꢀd6, ppm): δ 49.9, 61.4, 61.9, 65.3, 67.0, 70.3, 71.0, 74.4,
100.2, 116.5, 118.3, 118.9, 119.3, 121.8, 122.5, 124.1, 125.4,
132.6, 142.9, 148.6, 155.6, 156.4, 162.7; HRMS: calcd for
C114H128N20O48Na+, 2568.8121; found 2568.8109.
Synthesis of compound 4: Compound 2 (500 mg, 0.8 mM),
K2CO3 (913 mg, 6.6 mM) and compound 3 (735 mg, 5.0 mM)
were mixed in NMP (40 mL). The reaction mixture was heated at
90 °C for 12 h. After cooling to room temperature, the reaction
25 mixture was poured into hydrochloric acid solution, the solid
compound was filtered, washed by water. The residue was
purified by silicaꢀgel column chromatography using
CH2Cl2/petroleum ether (v/v = 10/1) as the eluent to give the
product as a red powder (350 mg) at yield of 40.2 %. m. p. 233.4ꢀ
30 235.6 °C. 1H NMR (600 MHz, CDCl3, ppm): δ 2.15 (t, J = 1.8 Hz,
2H, ꢀC≡CH); 2.57 (t, J = 1.8 Hz, 4H, ꢀC≡CH); 4.67 (d, J = 2.4
Hz, 8H, ꢀOCH2ꢀ); 4.88 (d, J = 1.8 Hz, 4H, ꢀNCH2ꢀ); 6.90 (s, 16H,
ArꢀH); 8.17 (s, 4H, peryleneꢀH); 13C NMR 150 MHz, CDCl3,
ppm): δ 29.7, 56.3, 70.6, 75.8, 78.3, 78.3, 116.4, 119.2, 119.4,
35 120.2, 121.2, 122.1, 132.9, 149.1, 154.6, 156.4, 162.5. HRMS:
calcd for C66H38N2O12, 1050.2425; found 1050.2435.
80 Acknowledgements
We thank NSFC (21002020), Hebei Natural Science Foundation
(B2011201052, B2012201041) and the Foundation of Hebei
Education Department (2010106) for financial support.
References
1
(a) D. H. Dube, C. R. Bertozzi, Nat. Rev. Drug Discovery, 2005, 4,
477; (b) Z. Shriver, S. Raguram, R. Sasisekharan, Nat. Rev. Drug
Discovery, 2004, 3, 863; (c) P. M. Rudd, T. Elliott, P. Cresswell, I. A.
Wilson, R. A. Dwek, Science, 2001, 291, 2370; (d) R. Raman, S.
Raguram, G. Venkataraman, J. C. Paulson, R. Sasisekharan, Nat.
Methods, 2005, 2, 817; (e) M. M. Fuster, J. D. Esko, Nat. Rev.
Cancer, 2005, 5, 526; (f) F. T. Liu, G. A. Rabinovich, Nat. Rev.
Cancer, 2005, 5, 29; (g) C. M. Szymanski, B. W. Wren, Nat. Rev.
Microbiol., 2005, 3, 225; (h) K. Ohtsubo, J. D. Marth, Cell, 2006, 126,
855; (i) P. R. Crocker, J. C. Paulson, A. Varki, Nat. Rev. Immunol.,
2007, 7, 255; (j) P. Niederhafner, M. Reiniš, J. Šebestík, J. Ježek, J.
Pept. Sci., 2008, 14, 556.
Synthesis of compound 6: Compound 4 (300 mg, 0.28 mmol)
and compound 5 (254 mg, 2.02 mM) were dissolved in THF (50
mL), and the water solution of CuSO4ꢂ5H2O (38 mg, 0.51 mM)
40 and sodium ascorbate (50 mg, 0.84 mM) were added, respectively.
The mixture was stirred for 10 h at 55 °C. The solvent was
evaporated under vacuum. The residue was purified by silicaꢀgel
column chromatography using CH2Cl2/CH3OH (v/v = 30/1) as the
eluent to give the product as a dark red solid (460 mg) at yield of
45 63.3 %. m. p. 108.1ꢀ109.8 °C; FT–IR (KBr) v [cm–1]: 1749, 1699,
2
(a) T. C. Shiao, R. Roy, New J. Chem., 2012, 36, 324; (b) M. Touaibia,
R. Roy, MiniꢀRev. Med. Chem., 2007, 7, 1270; (c) R. J. Pieters, Med.
Res. Rev., 2007, 27, 796; (d) M. Hartmann, T. K. Lindhorst, Eur. J.
Org. Chem., 2011, 3583; (e) S. Jin, Y. Cheng, S. Reid, M. Li, B.
Wang, Med. Res. Rev., 2010, 30, 171.
3
4
(a) J. J. Lundquist, E. J. Toone, Chem. Rev., 2002, 102, 555; (b) M.
Mammen, S. K. Choi, G. M. Whitesides, Angew. Chem. Int. Ed., 1998,
37, 2754.
(a) A. Dondoni, Chem. Asian J., 2007, 2, 700; (b) D. Deniaud, K.
Julienne, S. G. Gouin, Org. Biomol. Chem., 2011, 9, 966; (c) O.
Renaudet, MiniꢀReview. Org. Chem., 2008, 5, 274 ; (d) A. Dondoni,
A. Marra, Chem. Rev., 2010, 110, 4949; (e) C. O. Mellet, J. Defaye, J.
M. G. Fernádez, Chem. Eur. J., 2002, 8, 1982.
1
1662, 1589, 1500; H NMR (600 MHz, CDCl3, ppm): δ 1.92 (s,
6H, ꢀCOCH3), 1.98 (s, 12H, ꢀCOCH3), 2.01 (s, 18H, ꢀCOCH3),
2.05 (s, 6H, ꢀCOCH3), 2.09 (s, 18H, ꢀCOCH3), 2.13 (s, 12H, ꢀ
COCH3), 3.52ꢀ3.57 (m, 6H), 3.82 (m, 2H), 3.92 (m, 4H), 3.98 (d,
50 J = 12.6 Hz, 2H), 4.03ꢀ4.07 (m, 6H), 4.13ꢀ4.22 (m, 10H), 4.50 (t,
J = 4.8 Hz, 4H), 4.65 (t, J = 5.4 Hz, 8H), 4.74 (s, 2H), 4.81 (s,
4H), 5.13 (s, 2H), 5.19ꢀ5.24 (m, 24H), 5.40ꢀ5.45 (m, 4H), 6.90
(dd, J = 10.8 Hz, 19.8 Hz, 16H, ArꢀH); 7.69 (s, 2H, ꢀTriaz1ꢀH);
7.80 (s, 4H, Triaz2ꢀH); 8.08 (s, 4H, peryleneꢀH); 13C NMR (150
55 MHz, CDCl3, ppm): δ 20.5, 20.6, 20.6, 20.7, 49.8, 62.2, 65.8,
66.2, 68.8, 69.1, 69.2, 97.5, 116.3, 119.3, 120.1, 121.4, 122.3,
124.0, 132.8, 144.1, 149.0, 155.3, 156.4, 162.9, 169.5, 169.8,
169.8, 170.4; HRMS: calcd for C162H176N20O72Na+, 3577.0657;
5
6
(a) A. VargasꢀBerenguel, F. OrtegaꢀCaballero, J. M. CasasꢀSolvas,
MiniꢀRev. Org. Chem., 2007, 4, 1; (b) R. J. Pieters, Org. Biomol.
Chem., 2009, 7, 2013; (c) Y. Lim, M. Lee, Org. Biomol. Chem.,
2007, 5, 401; (d) K. Li, B. Liu, Polym. Chem., 2010, 1, 252; (e) N.
Jayaraman, Chem. Soc. Rev., 2009, 38, 3463.
(a) M. K. Müller, L. Brunsveld, Angew. Chem. Int. Ed., 2009, 48,
2921; (b) J. H. Ryu, E. Lee, Y. Lim, M. Lee, J. Am. Chem. Soc.,
2007, 129, 4808; (c) B. S. Kim, D. J. Hong, J. Bae, M. Lee, J. Am.
Chem. Soc., 2005, 127, 16333; (d) B. S. Kim, W. Y. Yang, J. H. Ryu,
Y. S. Yoo, M. Lee, Chem. Commun., 2005, 2035.
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 | 5

Page 7 of 7
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View Article Online
7
8
9
M. P. Leal, M. Assali, I. Fernández, N. Khiar, Chem. Eur. J., 2011, 17,
1828.
K. Aplander, M. Marttila, S. Manner, N. Arnberg, O. Sterner, U.
Ellervik, J. Med. Chem., 2011, 54, 6670.
K. Petkau, A. Kaeser, I. Fischer, L. Brunsveld, A. P. H. Schenning, J.
Am. Chem. Soc., 2011, 133, 17063.
10 K. R. Wang, H. W. An, L. Wu, J. C. Zhang, X. L. Li, Chem. Commun.,
2012, 48, 5644.
11 (a) L. Zang, Y. Che, J. S. Moore, Acc. Chem. Res., 2008, 41, 1596; (b)
E. Krieg, B. Rybtchinski, Chem. Eur. J., 2011, 17, 9016; (c) D. Görl,
X. Zhang, F. Würthner, Angew. Chem. Int. Ed., 2012, 51, 6328.
12 M. Srinivasan, S. Sankararaman, H. Hopf, I. Dix, P. G. Jones, J. Org.
Chem., 2001, 66, 4299.
13 (a) J. Ni, S. Singh, L. X. Wang, Bioconjug. Chem., 2003, 14, 232; (b)
M. D. Disney, P. H. Seeberger, Chem. Biol., 2004, 11, 1701; (c) A. V.
Demchenko, G. J. Boons, J. Org. Chem., 2001, 66, 2547.
14 (a) W. Wang, W. Wan, H. H. Zhou, S. Niu, A. D. Q. Li, J. Am.
Chem. Soc., 2003, 125, 5248; (b) R. O. Marcon, J. G. dos Santos, K.
M. Figueiredo, S. Brochsztain, Langmuir, 2006, 22, 1680 ; (c) Z. J.
Chen, B. Fimmel, F. Würthner, Org. Biomol. Chem., 2012, 10, 5845.
15 T. E. Kaiser, H. Wang, V. Stepanenko, F. Würthner, Angew. Chem. Int.
Ed., 2007, 46, 5541.
16 A. D. Q. Li, W. Wang, L. Q. Wang, Chem. Eur. J., 2003, 9, 4594.
17 (a) F. Würthner, Z. Chen, F. J. M. Hoeben, P. Osswald, C. C. You, P.
Jonkheijm, J. V. Herrikhuyzen, A. P. H. J. Schenning, P. P. A. M.
van der Schoot, E. W. Meijer, E. H. A. Beckers, S. C. J. Meskers, R.
A. J. Janssen, J. Am. Chem. Soc., 2004, 126, 10611; (b) V. Dehm, Z.
Chen, U. Baumeister, P. Prins, L. D. A. Siebbeles, F. Wü1rthner,
Org. Lett., 2007, 9, 1085; (d) Z. Huang, E. Lee, H. J. Kim, M. Lee,
Chem. Commun., 2009, 6819; (e) J. C. Hu, W. F. Kuang, K. Deng, W.
J. Zou, Y. W. Huang, Z. X. Wei, C. F. J. Faul, Adv. Funct. Mater.,
2012, 22, 4149.
18 R. V. Vico, J. Voskuhl, B. J. Ravoo, Langmuir, 2011, 27, 1391.
19 C. W. Cairo, J. E. Gestwicki, M. Kanai, L. L. Kiessling, J. Am.
Chem. Soc., 2002, 124, 1615.
20 (a) N. Kamiya, M. Tominaga, S. Sato, M. Fujita, J. Am. Chem. Soc.,
2007, 129, 3816; (b) R. Kikkeri, L. H. Hossain, P. H. Seeberger,
Chem. Commun., 2008, 2127; (c) S. Schmid, A. Mishra, P. Bäuerle,
Chem. Commun., 2011, 47, 1324.
21 R. Euzen, J. L. Reymond, Mol. BioSyst., 2011, 7, 411.
6
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