Design and Function of Supramolecular Recognition Systems Based on Guest-Targeting Probe-Modified Cyclodextrin Receptors for ATP

Article abstract of DOI:10.1021/acs.joc.6b02513

In this study, we have developed a rational design strategy to obtain highly selective supramolecular recognition systems of cyclodextrins (CyDs) on the basis of the lock and key principle. We designed and synthesized dipicolylamine (dpa)-modified -CyD-Cu

Full text of DOI:10.1021/acs.joc.6b02513

Subscriber access provided by Fudan University
Article
Design and Function of Supramolecular Recognition Systems Based
on Guest Targeting Probe-Modified Cyclodextrin Receptors for ATP
Kyohhei Fujita, Shoji Fujiwara, Tatsuru Yamada, Yuji Tsuchido, Takeshi Hashimoto, and Takashi Hayashita
J. Org. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.joc.6b02513 • Publication Date (Web): 20 Dec 2016
Downloaded from http://pubs.acs.org on December 22, 2016
Just Accepted
“Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted
online prior to technical editing, formatting for publication and author proofing. The American Chemical
Society provides “Just Accepted” as a free service to the research community to expedite the
dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts
appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been
fully peer reviewed, but should not be considered the official version of record. They are accessible to all
readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered
to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published
in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just
Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor
changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers
and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors
or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
The Journal of Organic Chemistry is published by the American Chemical Society.
1155 Sixteenth Street N.W., Washington, DC 20036
Published by American Chemical Society. Copyright © American Chemical Society.
However, no copyright claim is made to original U.S. Government works, or works
produced by employees of any Commonwealth realm Crown government in the course
of their duties.

Page 1 of 7
The Journal of Organic Chemistry
1
2
3
4
5
6
7
8
9
Design and Function of Supramolecular Recognition Systems Based
on Guest Targeting Probe-Modified Cyclodextrin Receptors for ATP
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
Kyohhei Fujita†,‡, Shoji Fujiwara†, Tatsuru Yamada†, Yuji Tsuchido†, Takeshi Hashimoto† and
Takashi Hayashita*†
†Department of Materials and Life Sciences, Faculty of Science and Technology, Sophia University, 7-1 Kioi-cho,
Chiyoda, Tokyo 102-8554, Japan
‡Graduate School of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo, Tokyo 113-0033, Japan
ABSTRACT: In this study, we have developed a rational
design strategy to obtain highly selective supramolecular
recognition systems of cyclodextrins (CyDs) on the basis
of the lock and key principle. We designed and synthe-
sized dipicolylamine (dpa)-modified
plexes possessing an azobenzene unit (Cu･1-
and examined how they recognized phosphoric acid
derivatives in water. The results revealed that Cu･1-
γ
-CyD-Cu2+ com-
-CyD)
γ
γ
-
CyD recognized ATP with high selectivity over other
phosphoric acid derivatives. The significant blue shift
1
in the UV-vis spectra and H NMR analysis suggested that the selective ATP recognition was based on the multipoint in-
teractions between the adenine moiety of ATP and both the CyD cavity and the azobenzene unit in addition to the recog-
nition of phosphoric moieties by the Cu-dpa complex site. Our unique receptor made it capable of distinguishing ATP
from AMP and ADP, revealing the discrimination of even a length of one phosphoric group. This study demonstrates
that, compared to conventional recognition systems of CyDs, this multipoint recognition system confers a higher degree
of selectivity for certain organic molecules, such as ATP, over their similar derivatives.
molecules.16-18 Until now, certain number of studies for
INTRODUCTION
the selective complexation of nucleotides and nucleosides
by synthetic CyD hosts have been reported.19-27 For in-
The supramolecular chemistry of molecular complexes
formed by weak interactions between host molecules and
stance, positively charged CyDs bearing some aminome-
guest molecules has been explored with the intent of de-
thyl groups bind adenosine monophosphate (AMP),
veloping new functional capabilities for highly selective
adenosine diphosphate (ADP) or adenosine triphosphate
molecular recognition systems.1-5 Cyclodextrins (CyDs)
(ATP) very tightly with interesting structural features, but
are well-known host molecules that include hydrophobic
there were no examples of applications to highly selective
chemical sensors.19,20,26 In the field of supramolecular
molecules and act to increase the solubilities of the hy-
drophobic guest molecules in water through the for-
chemistry, the relationship between CyDs and azoben-
mation of host-guest inclusion complexes.6,7 Molecular
zene derivatives has been well studied over recent dec-
ades.9,18,28 It is known that azobenzene-modified CyDs
recognition systems of CyDs based on their host-guest
interactions have caught a great deal of attention in a
provide supramolecular recognition systems possessing
variety of chemical fields. 8-10 A various functional CyDs
tor.9,18,29 However, the potential molecular targets of the-
have been studied as versatile receptors for molecular
the potential for a guest responsive color-change indica-
recognitions, building blocks for functional materials, and
se CyD probes are limited because the detection mecha-
even drug delivery systems.11-15 However, these recogni-
nisms depend only on the affinity of guest molecules to
tion systems have several limitations. For example, it is
CyDs cavity. In this research, in order to overcome these
quite difficult for them to selectively include certain or-
limitations, we demonstrate the utility of a novel ap-
ganic molecules over their similar derivatives because
proach in the design of supramolecular CyD recognition
most conventional recognition systems of CyDs are based
on 1 : 1 type interactions between CyD cavity and guest
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 2 of 7
systems based on the lock and key principle by imple-
menting selective colorimetric receptors for ATP.
obtained and identified by 1H NMR, 13C NMR, 1H-1H COSY
experiments, and ESI-HRMS (See Experimental Section).
1
2
3
4
5
6
7
8
9
Our receptor (Figure 1) possesses two recognition sites:
the dipicolylamine (dpa)-modified azobenzene unit that
appears like a protruding arm (guest targeting probe),
At first, the UV-vis spectra around the π-π* transition of
Cu･1 possessing no CyD cavity in the presence of mono-
phosphate (Pi), pyrophosphate (PPi), triphosphate (Tri),
AMP, ADP, and ATP were measured. No remarkable se-
lectivity for certain phosphoric acid derivatives was ob-
served in both the absorbance changes and the wave-
length shifts (Figure 2). However, a small blue shift was
observed in the presence of ATP.
and the γ-CyD cavity. The length of the arm was designed
to provide multipoint recognition systems by host-guest
interactions between the adenine moiety of ATP and the
CyD cavity, in addition to the recognition of phosphoric
moieties by the metal ion-dpa complex site. The dpa unit
is well known ligand for the recognition of phosphoric
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
moieties, and γ-CyD has the adequate cavity size to pro-
vide the host-guest interactions with adenine moiety of
ATP even after the modification of guest targeting probes.
We evaluated the recognition capabilities toward several
phosphoric acid derivatives of 1 and 1-γ-CyD (Scheme 1)
metal ion complexes.
Figure 1. Images of CyD probe for ATP based on host–
guest interactions.
Figure 2. Selectivity of Cu･1 in the presence of phos-
phoric acid derivatives in 1% DMSO-99% water (v/v), pH
7.4 adjusted with HEPES/NaOH buffer, at 25 ºC. [1] =
0.020 mM, [Cu(NO3)2] = 0.020 mM, [phosphoric acid de-
Scheme 1. Synthesis of dipicolylamine-modified CyD
rivative] = 2.0 mM, [γ-CyD] = 0.020 mM.
In order to increase selectivity and versatility of the recep-
tor, we not only focused on the affinity of the CyD cavity
to the guest but also the length of the arm, thereby allow-
ing for a potential range of guest lengths. To confirm the
validity of our unique recognition strategy based on guest
targeting probe-modified CyDs, we chose ATP as a target
molecule, because it is difficult for conventional host-
guest interactions of CyDs to selectively include such
complicated molecules over their similar derivatives.
RESULTS AND DISCUSSION
The probe 1 was synthesized by azo coupling30 and the
Figure 3. Selectivity of Cu･1-
phosphoric acid derivatives in 1% DMSO-99% water (v/v),
pH 7.4 adjusted with HEPES/NaOH buffer, at 25 ºC. [1-
γ-CyD in the presence of
Mannich reaction. Then, the probe 1 and 3-amino-γ-CyD
γ
-
were coupled by a condensation reaction.31 The crude
product was purified by acetone reprecipitation and gel
CyD] = 0.020 mM, [Cu(NO3)2] = 0.020 mM, [phosphoric
acid derivative] = 2.0 mM.
filtration chromatography.18,29 The probe 1-
γ-CyD was
2
ACS Paragon Plus Environment

Page 3 of 7
The Journal of Organic Chemistry
In order to evaluate the recognition capability of the
CyD cavity, the UV-vis spectra of Cu･1- -CyD in the
presence of phosphoric acid derivatives were also exam-
transition. As conventional spectral shifts of azobenzene
1
2
3
4
5
6
7
8
derivatives are due to photoisomerization or protona-
tion/deprotonation of the functional groups, this is a
unique response mechanism based on π-π interactions for
azobenzene derivatives.9,15
γ
ined. Cu･1-γ-CyD showed a significant blue shift only in
the presence of ATP (Figure 3). The Cu2+ complex exhib-
ited the most remarkable response to ATP, compared to
that of other metal ions such as Zn2+.
In addition, the results demonstrated that the anion se-
lectivity of Cu･1-γ-CyD was altered by the length of
phosphoric anion moieties and the existence of adenine
moieties. This selectivity was considered to be based on
host-guest interactions between the adenine moiety of
9
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
ATP and the γ-CyD cavity. It was assumed that the hy-
drophobic CyD cavity induced stronger π-π interactions
by immobilizing the adenine moiety in close proximity to
the azobenzene unit, compared to the Cu･1 recognition
system. The adenine moieties of both AMP and ADP did
not interact with the azobenzene unit because the lengths
of their phosphate anion moieties were inadequate for
interacting with the CyD cavity and the azobenzene unit.
It is evident that the distance between the dpa unit and
the CyD cavity plays an important role in the selective
ATP recognition in this system.
To evaluate the real selectivity of Cu･1-γ-CyD for ATP,
the competitive binding experiments are conducted in the
presence of other interfering anions (Pi, PPi, Tri, AMP,
and ADP), with the subsequent addition of ATP (Figure
5).32 The Cu･1-
γ
-CyD also showed blue shifts derived
from ATP recognition in the presence of interfering ani-
ons, indicating that Cu･1- -CyD recognized ATP selec-
γ
tively in responses even if other similar derivatives exist-
ed (The UV-vis spectra and the absorbance changes are
depicted in Figure S25).
Figure 4. The curve of molar absorption coefficients of
Cu･1-
water-1% DMSO (v/v), pH 7.4 adjusted by HEPES/NaOH
buffer, at 25ºC. [1- -CyD] = 0.020 mM, (i) in the absence
γ-CyD in the absence and presence of ATP in 99%
γ
of ATP, (ii) [ATP] = 2.0 mM.
To analyze this ATP-selective blue shift, each spectrum
was fitted by a Gaussian function (Figure 4). The fitting
results revealed that the UV-vis spectra were composed of
two optical transitions: an optically allowed π-π* transi-
tion and an optically forbidden n-π* transition. In partic-
ular, the oscillator strength of the n-π* transition was
increased in the presence of ATP (Tables S5 and S6).
The UV-vis spectra of the π-π* transition also broadened
and shifted to the high-energy region. The results
demonstrated that the π-π interactions between the ade-
nine moiety and the azobenzene unit were affected by the
spectral shift. In general, the energy gap between HOMO
(corresponding to π orbitals) and LUMO (corresponding
to π* orbitals) is enlarged by the formation of a more sta-
ble complex, and the absorption maximum wavelength
shifts to the high-energy region (It is known that adeno-
sine moiety easily interacts with other aromatic com-
pounds).12,32 Therefore, it was suggested that π-π interac-
tions induced a change in the electronic states around the
dye moiety and slightly broke the optically forbidden n-π*
Figure 5. Real selectivity of Cu･1-γ-CyD for ATP in
wavelength shifts toward anions in 99% water-1% DMSO
(v/v), pH 7.4 adjusted by HEPES/NaOH buffer, at 25ºC.
[1-γ-CyD] = 0.020 mM, [Cu(NO3)2] = 0.020 mM, [ATP] =
[Anion] = 2.0 mM. The gray bar represents the wave-
length shifts in the presence of each anion (100 equiv.).
The black bar represents wavelength shifts in the pres-
ence of ATP (100 equiv.) and each anion (100 equiv.).
3
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 4 of 7
In order to calculate the binding constant to ATP, the
absorption spectra of 1 and 1- -CyD metal ion complexes
at pH 7.4 in the presence of various concentrations of
ATP were examined (Figures S21-S24). The binding con-
stant of Cu･1 to ATP was 2630 ± 310 M-1 by curve fitting.
In addition, the shifts of the proton peaks assigned to a, b,
1
2
3
4
5
6
7
8
and d, which were derived from the dpa unit, implicated
the recognition of phosphoric moieties by the Zn-dpa
complex site.35-37 The shift of the proton peak assigned to
h, which was derived from the proton of the hydroxyl
group at the ortho position, was also noted, indicating
that the hydroxyl group is related to the coordination to
Zn2+ as a phenolate ion.
γ
On the other hand, that of Cu･1-γ-CyD was 6640 ± 890
M-1, which was 2.5 times higher than that of Cu･1 having
no CyD. Also, the binding constant of Zn･1 to ATP was
857 ± 119 M-1. On the other hand, that of Zn･1-
γ-CyD was
In NOESY measurements, protons that are in close
proximity to each other give rise to correlation peaks.33,34
According to Figure 7 (i), there are correlations between
1H derived from the dpa-azobenzene unit and H3 derived
from the inside cavity of CyD in the presence of one
equivalent of ATP (enclosed in blue circles). The result
suggested that conformations including and excluding
CyD-dpa-azobenzene unit exist in equilibrium in solu-
tion.18 However, in the presence of five equivalents of
ATP, correlations between α and β protons derived from
the adenine moiety and H3 derived from the inside cavity
of CyD (enclosed in red circles) were observed, and the
former correlation disappeared, as shown in Figure 7 (ii).
However, there were no correlations between α and β
protons and H5 derived from the inside cavity of CyD.
Therefore, it is evident that the adenine moiety interacts
with the inside cavity of CyD shallowly.27,34
9
2898 ± 146 M-1, which was 3.4 times higher than that of
Zn･1 having no CyD. These results confirmed that the
CyD cavity enhances the binding constant to ATP in this
recognition system.
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
We also analyzed the supramolecular conformation of
the Cu･1-γ
-CyD/ATP complex by conducting the 1H NMR
titration and NOESY measurements of the Zn･1-γ-CyD
/ATP complex (Cu2+ is not appropriate for NMR meas-
urements because of its paramagnetism). The peaks of
1
NMR spectra were identified by H-1H COSY experiments
(See SI). In the NMR titration, proton peaks assigned to j
1
and k in the H NMR spectra were shifted to the high
magnetic field region in the presence of ATP (Figure 6),
which confirmed that π-π interactions with the adenine
moiety increased the electron density around the azoben-
zene unit.
1
Figure 6. H NMR spectra of 1-
γ
-CyD, Zn･1-
γ
-CyD and
Figure 7. NOESY spectra of Zn･1-γ-CyD/ATP complex
Zn ･1-γ-CyD/ATP complex (500 MHz, solvent: 33%
(500 MHz, solvent: 50% DMSO-d6-50% D2O (v/v)). [1-
γ
-
DMSO-d6-67% D2O). [1-γ-CyD] = 0.50 mM, (i) in the ab-
sence of Zn2+ and ATP, (ii) [Zn(NO3)2] = 0.50 mM, (iii)
[Zn(NO3)2] = 0.50 mM, [ATP] = 50 mM.
CyD] = [Zn(NO3)2] = 5.0 mM, (i) [ATP] = 5.0 mM, (ii)
[ATP] = 25 mM.
4
ACS Paragon Plus Environment

Page 5 of 7
The Journal of Organic Chemistry
ATP with high selectivity over other phosphoric acid de-
These correlations were even observed in 50% DMSO-d6-
1
2
3
4
5
6
7
8
50% D2O (v/v) condition, which we can expect much
stronger interactions in 1% DMSO-99% water (v/v) condi-
rivatives possessing similar phosphate anion moieties or
the same nucleobase. The significant blue shift in the
UV-vis spectra and NMR analysis suggested that this se-
lectivity was based on the multipoint interactions be-
tween the adenine moiety of ATP and both the CyD cavity
and the azobenzene unit in addition to the recognition of
phosphoric moieties by the Cu-dpa complex site. These
results, which confirmed the successful binding of ATP to
tion. Also, if we could measure NMR spectra of Cu･1-γ-
CyD, much stronger correlations would be observed be-
cause Cu2+ complex showed the higher binding ability to
ATP (See Table S7). This important piece of evidence
supported that host-guest interactions occurred between
the adenine moiety of ATP and the CyD cavity.
9
Cu･1-γ-CyD, provided evidence that the distance be-
These results suggested the supramolecular confor-
mation shown in Figure 8. Thus, the host-guest interac-
tions between the adenine moiety of ATP and the CyD
cavity are responsible for the highly selective ATP recog-
nition in this supramolecular system. Our results confirm
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
tween the dpa unit and the CyD cavity plays an important
role in the selective ATP recognition. Thus our unique
recognition system made it capable of distinguishing ATP
from AMP and ADP, revealing the discrimination of even
a length of one phosphoric group. This supramolecular
recognition system based on the lock and key principle
would enable us to design novel highly selective chemical
receptors of CyDs for various kinds of organic molecules
in water.
the successful binding of ATP to the Cu･1-γ-CyD, sug-
gesting that, compared to conventional recognition sys-
tems of CyD―which were based on 1 : 1 type interactions
between the CyD cavity and guest molecules and not suf-
ficient for selective ATP recognition―the present mul-
tipoint recognition system confers a higher degree of se-
lectivity for certain organic molecules, such as ATP, over
their similar derivatives. These results also suggested that,
in order to increase selectivity and versatility of the recep-
tor, it is important not only to focus on the affinity of the
CyD cavity to the guest but also the distance between the
CyD cavity and the other recognition site, thereby allow-
ing for a potential range of guest lengths. By introducing
this strategy into the design of selective host-guest inter-
actions of CyDs, unprecedented selectivity and specificity
not observed with conventional CyD systems can be ex-
pected. The present recognition system is very simple but
provides a real model for the design of artificial receptors
based on the lock and key principle.
EXPERIMENTAL SECTION
Reagents. All organic solvents and reagents were com-
mercially available with guaranteed grades and used
without further purification. Water was doubly distilled
and deionized by a Milli-Q water system before use.
Apparatus. UV-vis absorption spectra were measured
with a UV-vis spectrophotometer equipped with a Peltier
thermocontroller with a 10 mm quartz cell.
Phosphoric acid derivatives recognition by Cu･1.
To evaluate the phosphoric acid recognition ability of
Cu･1, spectral measurements were performed. Solutions
containing 1 (0.020 mM), Cu(NO3)2 (0.020 mM), phos-
phoric acid derivatives (2.0 mM, pH 7.4, adjusted using
NaOH), γ-CyD (0.020 mM), and HEPES/NaOH buffer (5.0
mM, pH 7.4, adjusted using NaOH) were prepared and
spectra were recorded at 25ºC.
Phosphoric acid derivatives recognition by Cu･1-
CyD. To evaluate the phosphoric acid recognition ability
of Cu･1- -CyD, UV-vis spectral measurements were per-
formed. Solutions containing 1- -CyD (0.020 mM),
γ-
γ
γ
Cu(NO3)2 (0.020 mM), phosphoric acid derivatives (2.0
mM, pH 7.4, adjusted using NaOH), and HEPES/NaOH
buffer (5.0 mM, pH 7.4, adjusted using NaOH) were pre-
pared and spectra were recorded at 25ºC.
Spectral fitting analysis. UV-vis spectra obtained from
phosphoric acid derivatives recognition were fitted by
Gaussian functions.
Figure 8. Suggested supramolecular conformation of
Cu･1-γ
-CyD/ATP complex.38,39
The competitive binding experiments of Cu･1-
CyD. To evaluate the real selectivity of Cu･1- -CyD,
spectral measurements were performed. Solutions con-
taining 1- -CyD (0.020 mM), Cu(NO3)2 (0.020 mM), ATP
γ-
γ
CONCLUSION
γ
We have designed, synthesized, and evaluated the guest
(2.0 mM, pH 7.4, adjusted using NaOH), interfering ani-
ons (2.0 mM, pH 7.4, adjusted using NaOH), and
HEPES/NaOH buffer (5.0 mM, pH 7.4, adjusted using
NaOH) were prepared and spectra were recorded at 25ºC.
targeting probe-modified CyD (Cu･1-
design strategy of supramolecular recognition systems of
CyDs. The results revealed that Cu･1- -CyD recognized
γ-CyD) as the novel
γ
5
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 6 of 7
7.80 (m, 3H), 7.46 (d, J = 8.3 Hz, 2H), 7.29 (td, J = 5.5 Hz,
Calculation of binding constants of 1 and 1-
metal ion complexes to ATP. To calculate the binding
constant of 1 and 1- -CyD metal ion complexes to ATP,
UV-vis spectral measurements were performed. Solutions
containing 1- -CyD (0.010 mM), Cu(NO3)2 or Zn(NO3)2
(0.010 mM), and HEPES/NaOH buffer (5.0 mM, pH 7.4,
adjusted using NaOH) were prepared and spectra were
recorded at 25ºC while ATP concentrations were varied
from 0.0 to 4.0 mM. The binding constants were calculat-
ed by curve fitting (See SI).
γ-CyD
1
2
3
4
5
6
7
8
7.2 Hz, 7.2 Hz, 2H), 7.00 (d, J = 8.8 Hz, 1H), 3.86 (s, 4H),
3.83 (s, 2H). 13C NMR (500 MHz, DMSO-d6): δ 166.8, 161.2,
158.2, 154.6, 148.7, 145.0, 136.9, 131.9, 130.5, 125.3, 124.7, 124.5,
122.7, 122.4, 122.1, 116.6, 58.5, 54.1. ESI-HRMS (-) m/z calcd.
for [M-H]-, 452.1728; found, 452.1750.
γ
γ
General Procedure for the Synthesis of 1-γ-CyD. A
mixture of 1 (0.0530 g, 0.116 mmol), DCC (0.0238 g, 0.115
mmol), HOBt・H2O (0.0177 g, 0.115 mmol) in 3.0 cm3 dry
DMF was stirred in an ice bath for 15 min. The compound
9
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
NOESY measurements of Zn･1-
To evaluate the supramolecular conformation of the Cu･
1- -CyD/ATP complex, NOESY spectral measurements of
Zn･1- -CyD/ATP complex were performed. Solutions
containing 1- -CyD (5.0 mM), Zn(NO3)2 (5.0 mM), and
γ-CyD/ATP complex.
3-amino-γ-CyD (0.100 g, 0.0770 mmol) dissolved in a
small amount of dry DMF was slowly added, the solution
was stirred in an ice bath for 30 min again. After remov-
ing an ice bath, the reaction mixture was stirred at room
temperature for 24 h. The solution was concentrated un-
der reduced pressure until a half of DMF was removed.
The sample was put in refrigerator and left a day. The
resulting solution was poured into acetone (1000 cm3) to
obtain yellow precipitations, followed by washing with
acetone several times. The crude product was charged on
a column of Sephadex G10 and eluted with 0.2 M aqueous
solution of ammonium carbonate (pH = 8.9). The first
yellow spot was collected as the desired product. The
purity was checked with TLC. The fraction was lyophi-
lized under reduced pressure to form a yellow powder.
γ
γ
γ
ATP (5.0 mM or 25 mM, pH 7.4, adjusted using NaOD)
were prepared and NOESY spectra were recorded.
General Procedure for the Synthesis of 4’-
Hydroxyazobenzene-4-carboxylic acid.40 The com-
pound p-aminobenzoic acid (2.034 g, 14.80 mmol) was
dissolved in 3 M HClaq. (50 cm3), the solution was stirred
for 10 min in an ice bath (pH = 1). Sodium nitrite (1.056 g,
14.49 mmol) dissolved in 6.0 cm3 cooled DI water was
added, the solution was stirred for 15 min. Phenol (1.406 g,
14.87 mmol) and 4.0 cm3 5 M NaOHaq. dissolved in 4.0
cm3 cooled DI water was slowly added, stirred for 15 min
in an ice bath at 0 ºC. The solution was adjusted to pH =
7 with 5 M NaOHaq. Sodium chloride (2.072 g) was added
and heated at 65 ºC in a hot bath for 10 min, cooled at
room temperature and salted it out. Yellow precipitation
was filtered by suction filtration. The crude product was
recrystallized from MeOH and water to obtain the desired
product 4’-Hydroxyazobenzene-4-carboxylic acid (2.248
g) as orange-red crystals in 64% yield. The sample was
The compound 1-γ-CyD (110 mg) was obtained in 82%
1
yield. The sample was identified by H NMR, 1H-1H COSY
1
experiments (See SI), and ESI-HRMS. mp 305-306 ºC; H
NMR (500 MHz, DMSO-d6): δ 8.55 (d, J = 5.2 Hz, 2H), 8.05
(d, J = 8.6 Hz, 2H), 7.91 (d, J = 8.6 Hz, 1H), 7.82 (d, J = 6.9
Hz, 2H), 7.72-7.82 (m, 3H), 7.46 (d, J = 8.0 Hz, 2H), 7.29
(td, J = 6.0 Hz, 7.2 Hz, 7.2 Hz, 2H), 7.01 (d, J = 8.6 Hz, 1H),
6.56 (s, 1H), 5.57-6.01 (m, 19 H), 4.39-5.07 (m, 22 H), 3.90-
3.10 (m, overlaps with HOD), 3.81 (s, 4H), 3.78 (s, 2H).
ESI-HRMS (-) m/z calcd. for [M-H]-, 1729.6019; found,
1729.6019.
1
1
identified by H NMR, 13C NMR and ESI-HRMS. H NMR
(300 MHz, DMSO-d6): δ 9.17 (s, 1H), 6.80 (d, J = 8.8 Hz,
2H), 6.56 (dd, J = 8.8 Hz, 9.9 Hz, 4H), 5.66 (d, J = 8.8 Hz,
2H). 13C NMR (400 MHz, DMSO-d6): δ 167.8, 161.9, 153.7,
145.1, 135.9, 130.3, 125.1, 121.6, 116.1. ESI-HRMS (-) m/z calcd.
for [M-H]-, 241.0619; found, 241.0627.
ASSOCIATED CONTENT
Supporting Information
The supporting information is available free of charge on
the ACS publication web site.
General Procedure for the Synthesis of 1. A mixture
of dipicolylamine (0.825 g, 4.13 mmol) and 37% formalde-
hyde solution (0.338 g, 4.13 mmol) in 15 cm3 MeOH was
Compound characterization data, phosphoric acid
recognition by UV-vis measurements of 1 and 1-γ-
CyD metal ion complexes, calculation of binding
reflexed for 1.5
h
at 70 ºC.
The compound 4’-
constants by curve fitting, competitive experiments
Hydroxyazobenzene-4-carboxylic acid (1.008 g, 4.13
mmol) dissolved in 30 cm3 MeOH was added, the reaction
was maintained at reflux temperature of 70 ºC for 24 h.
The solution was concentrated under reduced pressure to
obtain orange-red viscous fluid. The fluid was washed
with small amount of MeOH and filtered by suction filtra-
tion to afford the desired product. The compound 1
(0.751 g) was obtained as a blight yellow solid in 40% yield.
1
and copies of H-1H COSY and NOESY spectra of
Zn･1-γ-CyD/ATP complex.
AUTHOR INFORMATION
Corresponding Author
*E-mail ta-hayas@sophia.ac.jp
1
The sample was identified by H NMR, 13C NMR, DEPT,
Author Contributions
1
and ESI-HRMS. mp 207 ºC; H NMR (300 MHz, DMSO-
All the authors have contributed equally to this work and
approved the final version of the manuscript.
d6): δ 8.54 (dd, J = 1.1 Hz, 5.0 Hz, 2H), 8.11 (d, J = 8.8 Hz,
2H), 7.92 (d, J = 5.1 Hz, 1H), 7.89 (d, J = 10.2 Hz, 2H), 7.77-
6
ACS Paragon Plus Environment

Page 7 of 7
The Journal of Organic Chemistry
(23) Cotner. E. S.; Smith, P. J. J. Org. Chem., 1998, 63,
1
2
3
4
5
6
7
8
Notes
1737-1739.
The authors declare no competing financial interests.
(24) Hauser, S. L.; Johanson, E. W.; Green H. P.; Smith, P.
J. Org. Lett., 2000, 16, 3575.
(25) Mourtzis, N.; Eliadou, K.; Aggelidou, C.; Sophianop-
oulou, V.; Mavridis, I. M.; Yannakopoulou, K. Org. Biomol.
Chem., 2007, 5, 125-131.
(26) Yuan, D.; Izuka, A.; Fukudome, M.; Rekharsky, M.
V.; Inoue, Y.; Fujita, K. Tetrahedron. Lett. 2007, 48, 3479-
3483.
(27) Aggelidou, C.; Mavridis, I. M.; Yannakopoulou, K.
Eur. J. Org. Chem., 2009, 2299-2305.
ACKNOWLEDGMENT
This work was financially supported by a Grant-in-Aid for
Scientific Research (C) (Grant No. 15K05548) from Japan So-
ciety for the Promotion of Science (JSPS) and a Grant-in Aid
for Scientific Research (A) (Grant No. 26248038) from the
Ministry of Education, Culture, Sports, Science and Technol-
ogy (MEXT) of Japan.
9
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
(28) Nonaka, K.; Yamaguchi, M.; Yasui, M.; Fujiwara, S.;
Hashimoto, T.; Hayashita, T. Chem. Commun., 2014, 50,
10059-10061.
(29) Kuwabara, T.; Shiba, K.; Nakajima, H.; Ozawa, M.;
Miyajima, N.; Hosoda, M.; Kuramoto, N.; Suzuki, Y. J.
Phys. Chem., 2006, 110, 13521-13529.
(30) Tsuchido, Y.; Aimu, K.; Toda, Y.; Hashimoto, T.;
Hayashita, T. J. Ion Exchange, 2014, 25(4), 146-150.
(31) Fujiwara, S.; Takahashi, K. Supramol. Chem., 2011,
23(1-2), 156-159.
(32) Xu, Z.; Singh, N. J.; Lim, J.; Pan, J.; Kim, H. N.; Park,
S.; Kim, K.; Yoon, J. J. Am. Chem. Soc., 2009, 131, 15528-
15533.
(33) Wang, J.; Pham, D.; Kee, T. W.; Clafton, S. N.; Guo,
X.; Clements, P.; Lincoln, S. F.; Prud’homme, R. K.; Easton,
C. J. Macromolecules, 2011, 44, 9782-9791.
(34) Wang, H.; Shao, N.; Qiao, S.; Cheng, Y. J. Phys.
Chem., 2012, 116, 11217-11224.
(35) Ojida, A.; Inoue, M.; Mito-Oka, Y.; Tsutsumi, H.;
Sada, K.; Hamachi, I. J. Am. Chem. Soc., 2006, 128, 2052-
2058.
(36) Rhee, H. W.; Lee, C. R.; Cho, S. H.; Song, M. R.;
Cashel, M.; Choy, H. E.; Seok, Y. J.; Hong, J. I. J. Am. Chem.
Soc., 2008, 130, 784-785.
(37) Ojida, A.; Sakamoto, T.; Inoue, M.; Fujishima, S.;
Lippens, G.; Hamachi, I. J. Am. Chem. Soc., 2009, 131,
6543-6548.
REFERENCES
(1) Hargrove, A. E.; Nieto, S.; Zhang, T.; Sessler, J. L.; An-
slyn, E. V. Chem. Rev., 2011, 111, 6603-6782.
(2) Joseph, R.; Rao, C. P., Chem. Rev., 2011, 111, 4658-4702.
(3) Kobayashi, H.; Hashimoto, T.; Hayashita, T. Synergy
in Supramolecular Chemistry, Chapter 13, CRC Press,
2015, pp.235-246.
(4) Lakkakula, S.; Mitkin, O. D.; Valiulin, R. A.;
Kutateladze, A. G. Org. Lett., 2007, 9, 1077-1079.
(5) Kurishita, Y.; Kojira, T.; Ojida, A.; Hamachi, I. J. Am.
Chem. Soc., 2010, 132, 13290-13299.
(6) Tellini, V. H. S.; Jover, A.; Garcia, J. C.; Galantini, L.;
Meijide, F.; Tato, J. V. J. Am. Chem. Soc., 2006, 128, 5728-
5734.
(7) Patra, D.; Zhang, H.; Sengupta, S.; Sen, A. ACS Nano,
2013, 7, 7674-7679.
(8) Wenz, G. Angew. Chem. Int. Ed. Engl. 1994, 33, 803-
822.
(9) Ueno, A.; Kuwabara, T.; Nakamura, A.; Toda, F. Na-
ture, 1992, 356, 136-137.
(10) Kumai, M.; Kozuka, S.; Samizo, M.; Hashimoto, T.;
Suzuki, I.; Hayashita, T. Anal. Sci., 2012, 28, 121-125.
(11) Liu, Y.; Chen, Y. Acc. Chem. Res., 2006, 39, 681-691.
(12) Chen, H. Y.; Zhao, M.; Li, Y.; Liu, G. F.; Ji, L. N.; Mao,
Z. W. Tetrahedron Lett., 2014, 55, 1802-1805.
(13) Tamura, A.; Yui, N. Sci. Rep., 2014, 4.
(14) Kralova, J.; Kejik, Z.; Briza, T.; Pouckova, P.; Kral, A.;
Martasek, P.; Kral, V. J. Med. Chem., 2010, 53, 128-138.
(15) Tamesue, S.; Takashima, Y.; Yamaguchi, H.; Shinkai,
S.; Harada, A. Angew. Chem. Int. Ed., 2010, 49, 7461-7464.
(16) Szente, L.; Szeman, J. Anal. Chem., 2013, 85, 8024-
8030.
(17) Liu, Y.; Shi, J.; Guo, D.; J. Org. Chem., 2007, 72, 8227-
8234.
(18) Kuwabara, T.; Sugiyama, K. Anal. Sci., 2013, 29, 905-
908.
(19) Eliseev, A. V.; Schneider, H. J. Angew. Chem. Int. Ed.,
1993, 32, 1331-1333.
(20) Eliseev, A. V.; Schneider, H. J. J. Am. Chem. Soc.,
1994, 116, 6081-6088.
(38) Szejtli, J.; Chem. Rev., 1998, 98, 1743-1753.
(39) Moniruzzaman, M.; Sabey, C. J.; Fernando, G. F.
Macromolecules, 2004, 37, 2572-2577.
(40) Poloni, C.; Szymanski, W.; Feringa, B. L.; Chem.
Commun., 2014, 50, 12645-12648.
(21) Schwinte, P.; Darcy, R.; Keeffe, F. J. Chem. Soc., Per-
kin Trans., 1998, 2, 805.
(22) Vizitiu, D.; Thatcher, G. R. J. J. Org. Chem., 1999, 64,
6235-6238.
7
ACS Paragon Plus Environment