Recognition of Viologen Derivatives in Water by N-Alkyl Ammonium Resorcinarene Chlorides

Article abstract of DOI:10.1021/acs.joc.7b00449

Three water-soluble N-alkyl ammonium resorcinarene chlorides decorated with terminal hydroxyl groups at the lower rims were synthesized and characterized. The receptors were decorated at the upper rim with either terminal hydroxyl, rigid cyclohexyl, or fl

Full text of DOI:10.1021/acs.joc.7b00449

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Article
The Recognition of Viologen Derivatives in Water
by N-Alkyl Ammonium Resorcinarene Chlorides
Ngong Kodiah Beyeh, Hyun Hwa Jo, Igor Kolesnichenko, FangFang Pan,
Elina Kalenius, Eric Van Anslyn, Robin H. A. Ras, and Kari Rissanen
J. Org. Chem., Just Accepted Manuscript • Publication Date (Web): 28 Apr 2017
Downloaded from http://pubs.acs.org on May 1, 2017
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The Recognition of Viologen Derivatives in Water by N-
Alkyl Ammonium Resorcinarene Chlorides
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Ngong Kodiah Beyeh,a* Hyun Hwa Jo,b Igor Kolesnichenko,b Fangfang Pan,c Elina Kalenius,d
Eric V. Anslyn,b* Robin H. A. Rasa and Kari Rissanend*
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a Aalto University, School of Science, Department of Applied Physics, Puumiehenkuja 2, FIꢀ02150 Espoo, Finland
b The University of Texas at Austin, Department of Chemistry, Austin, Texas 78712, USA
c Central China Normal University, College of Chemistry, Key Laboratory for Pesticides and Chemical Biology, 152 Luoyu
Road, Wuhan, Hubei 430079, China
d University of Jyvaskyla, Department of Chemistry, Nanoscience Centre, P. O. Box 35, FIꢀ40014 Jyvaskyla,
Finland
Supporting Information Placeholder
[
]
ABSTRACT: Three waterꢀsoluble Nꢀalkyl ammonium resorcinarene chlorides decorated with terminal hydroxyl
groups at the lower rims were synthesized and characterized. The receptors were decorated at the upper rim with
either terminal hydroxyl, rigid cyclohexyl or flexible benzyl groups. The binding affinities of these receptors
towards three viologen derivatives, two of which possess an acetylmethyl group attached to one of the pyridine
1
nitrogens, in water were investigated via H NMR spectroscopy, fluorescence spectroscopy, and isothermal titration
calorimetry (ITC). ITC quantification of the binding process gave association constants of up to 103 Mꢀ1. Analyses
reveal a spontaneous binding process which are all exothermic, and are both enthalpy and entropy driven.
INTRODCUTION
Molecular receptors with preꢀorganized cavities suitable for guest recognition is a continuously developing area in
supramolecular chemistry, material science, and biology.1,2. Receptors capable of guest recognition in aqueous media and
biological fluids have a growing importance, relative to receptors that primarily function in organic media.3–6 However,
receptors that can operate in aqueous media have proven to be difficult to design.7–12 However, they have potential
biocompatibility if one can exploit the cohesive force of water.3–6
Viologen (1,1’ꢀdisubstitutedꢀ4,4’ꢀbipyridine salts) materials are well known for their strong redox properties.13
Viologens, with their extended πꢀconjugation, can exhibit excellent electrochromic and photochromic properties.14–16
These species are commonly used in supramolecular chemistry to construct capsular assemblies or threaded structures
with several host compounds.17–21 Their cationic nature makes them suitable guests for πꢀrich receptors.17–21 The
complexation of viologen derivatives, by receptors such as curcubiturils, can substantially alter the kinetics and
thermodynamics of their electron transfer reactions.17,18 Modified viologens have potential as sensors for bisulfite in water
which can be utilized in the beverage industry.22–25
NꢀAlkyl ammonium resorcinarene halides (NARXs), resulting from the ring opening of tetrabenzoxazines in the
presence of mineral acids under refluxing conditions, are stabilized by a seam of hydrogenꢀbonded cationꢀanion
interactions.26,27 The NARX receptors possess four spatially fixed halide anions with deep cavities for guest binding.
Neutral and anionic guests have been shown to reside in the cavity of NARXs, interacting with the host mainly through
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CHꢀπ interactions and hydrogen bonds.28,29 We recently reported the binding of small neutral molecules such as amides
and diamides in organic media with cooperativity by NARX receptors.30 The four spatially fixed anions act as halogen
bond acceptors leading to a variety of complex architectures, such as deepꢀcavity cavitands, pseudoꢀcapsular, and capsular
assemblies.31,32 The NARXs are generally soluble in alcoholic and nonꢀpolar solvents. Recently we synthesized the first
water soluble NARX receptors by attaching terminal hydroxyl groups at the upper rim.33 Therein, we showed that the
water soluble NARXs exist in C4v crown conformation and bind a variety of aliphatic, aromatic and halogenated alkanes
and arenes in aqueous media.33
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In this study, three new waterꢀsoluble NARCl receptors decorated with four terminal hydroxyl groups at the lower rims
were synthesized and characterized (Figure 1). The first NARCl receptor (5), is also functionalized at the upper rim with
four terminal hydroxyl groups, making it extremely water soluble (35 mg/mL). The receptor (7) is functionalized at the
upper rim with four rigid cyclohexyl groups and the third receptor (9) possesses four flexible benzyl groups at the upper
rim. Monoꢀmethyl 4,4’ꢀbipyridine (11),34 monoꢀacetylmethyl 4,4’ꢀbipyridine (12), and hetero methylꢀacetylmethyl 4,4’ꢀ
bipyridine (13) molecules were also synthesized as potential guests. The recognition of these guests (11ꢀ13) by the NARX
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receptors (5, 7 and 9) were investigated in water via H NMR spectroscopy, fluorescence spectroscopy, and isothermal
titration calorimetry (ITC) analyses.
RESULTS AND DISCUSSION
Synthesis of the receptors and the viologen guests. The synthesis of 5, 7 and 9 starts with resorcinarene 3, which was
synthesized through reported procedures.35 Ethanolamine, cyclohexyl amine and benzyl amine in the presence of excess
formaldehyde participate in a Mannich condensation with 3 to form tetrabenzoxazines 4, 6 and 8, respectively.36,37 The
reaction with ethanolamine leads to a mixture of the five and six membered azoxazine rings 4a and 4b, respectively. The
unꢀisolated crude product containing the fiveꢀ and sixꢀmembered ring compounds in the presence of concentrated HCl
under refluxing conditions, lead to the same final product 5 (Figure 1). Cleavage of the pure tetrabenzoxazines 6 and 8
under similar conditions give NARCls 7 and 9. The detailed synthetic procedures of the NARCl receptors are reported in
the Supporting Information (Schemes S1ꢀS3; Figures S1ꢀS5).
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Synthesis of Hosts
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OH
OH
O
O
N
NH2
N
Cl
HCl,
H2O
HO
OH
HO
OH
HO
i-PrOH
reflux, 4 h
4
4
4
4b
4a
HO
5
HO
HO
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CH2O
HOC2H4NH2
EtOH,r.t., 24 h
N
NH2
Cl
C6H11NH2
HCl, H2O
HO
OH
CH O
2
HO
O
HO
OH
EtOH
r.t., 24 h
i-PrOH
reflux, 4 h
4
4
4
3
6
7
HO
HO
HO
C7H7NH2
CH2O
EtOH
r.t., 24 h
HCl,H2O
EtOH,0-65 oC
N
NH2
Cl
7 days
HCl, H2O
HO
O
HO
OH
HO
OH
O
i-PrOH
reflux, 4 h
+
4
4
8
9
1
2
HO
HO
Synthesis of Guests
O
O
Cl
N
N
N
N
Cl
O
10
THF
reflux
12
MeI
CH2Cl2
O
Cl
N
N
N
N
MeCN
reflux
Cl
I
11
I
13
Figure 1. Synthesis of water soluble NARCls 5, 7 and 9 as receptors and derivatives of 4,4’ꢀbipyridine 11ꢀ13 as guests.
The 4,4’ꢀbipyridine guest 11 was synthesized according to a reported procedure.34 The other bipyridine guests, 12 and 13,
were synthesized by reacting chloroacetone with 4,4’ꢀbipyridine or guest 11, respectively (Schemes S4, S5; Figures S6ꢀ
S8). Suitable single crystals of monoꢀmethyl 4,4’ꢀbipyridine (11) and heteroꢀ methylꢀacetylmethyl 4,4ꢀbipyridine (13)
were obtained and analyzed (Figures S9ꢀS11). Structural analysis verified the bipyridine was successfully substituted. In
11, the Nꢀmethyl 4,4’ꢀbipyridyl cation is paired with the iodide anion with the anion close to the cationic nitrogen of the
viologen molecule. While in 13, although the dicationic viologen is as expected, the original counter anions are replaced
by 1.5 Iꢀ and 0.5 I3 during the crystallization. Electrostatic forces contribute to the arrangement of the ion pairs. A
ꢀ
mechanism for formation of the triiodide from iodide was proposed in a recent crystallographic study.38
Hydrogen/Deuterium (H/D) Exchange Studies of the Viologen Guests. In D2O, the labile ꢀNCH2COꢀ hydrogens
undergo H/D exchange. These protons are therefore not observed in the H NMR spectra of all samples containing guests
12 and 13 in protic deuterated solvents. The lability of these hydrogens was verified by electrospray ionization mass
spectrometry (ESIꢀMS) in a combined experiment of solution H/Dꢀexchange and collision induced dissociation (CID). In
D2O (Figure 2, Figures. S12, S13), two H/Dꢀexchanges of ꢀNCH2COꢀ hydrogens were observed. The location of
exchanged hydrogens was verified by CID experiment, which showed the fragmentation to be initiated by elimination of
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undeuterated acetyl radical (C2H3O•, 43 u), leaving only one plausible location for H/Dꢀexchange at ꢀNCH2COꢀ. The CID
experiments also showed the increased stability of keto tautomer as compared to possible enol form. This is likely due to
existence of two resonance structures of keto tautomer (see SI, Figure S12).
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Figure 2. ESIꢀMS of 12 in D2O. (a) Profile spectrum showing two H/Dꢀexchanges, inset showing zoomed view for m/z
215. (b) CID for isolated [12D2]+ ion (CE=29). (c) Main fragmentation pathway for ion [12D2]+.
Complexation Studies via NMR Spectroscopy. Complexation studies between the NARCl receptors (5, 7 and 9) with
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the modified 4,4ꢀbipyridine guests (11ꢀ13) were investigated by H NMR experiments in D2O. The receptors possess C4v
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symmetry in solution as observed from their relatively simple H NMR spectra (Figures S1, S3 and S5). Varying
complexationꢀinduced shift changes of the different guest signals were observed from either the shielding effects of the
aromatic rings of the bowlꢀshaped host cavity or interaction with the cationꢀanion seam. The complexation process is fast
on the NMR time scale at 298 K. The guests are soluble in water and can interact with the hosts mainly through CHꢀπ and
hydrogen bond interactions with the cationꢀanion seam of the receptors. The small shift changes of the guests can be
attributed to the highly competitive nature of the bulk water. Taking the complex 13@7 as an example, upfield shifts of
the guest –COCH3ꢀ protons are observed (Figure 3). The carbonyl oxygens are known to interact with the cationꢀanion
seam in organic media as reported in the binding of amides by NARXs.30 Upfield shift of methyl protons of ammonium
cations are usually observed when located deep in the cavity of resorcinareneꢀtype receptors. The relatively small upfield
shifts of the methyl protons of guest 13 therefore suggest the binding interaction to occur mainly at the upper rim of the
receptors involving the hydrogenꢀbonded cationꢀanion seam with minimal interaction with the electron rich interior cavity
of the resorcinarene cavity (Figure 3).
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Figure 3. Selected region of the H NMR (400 MHz, D2O, 298 K) spectra observed upon the addition of the guest 13 to
the host 7. Dotted lines gives an indication of the complexation induced shift changes.
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The 1H NMR spectra of the 1:1 mixture between the guests 11 and 12 reveal mainly downfield shifts of the guests signals
(Figures S15ꢀS20). Limited and/or no shift changes were observed for the guests methyl signals. This again suggest the
interaction between the guests 11 and 12 with the receptors to be mainly at the upper rim of the receptors involving the
hydrogenꢀbonded cationꢀanion seam of the receptors and the free pyridine nitrogens of the guests (Figures S15ꢀS22). Such
hydrogen bond interactions between the pyridine nitrogen of the guests (11 and 12) leads to deꢀshielding contrary to the
guest 13 with no free pyridine nitrogen.
Quantification of the Binding Process via Isothermal Titration Calorimetry (ITC) Studies. The interaction between
the hosts and guests was quantified through a series of ITC experiments in H2O (Figure 4, Figures S23, S24). The
thermodynamic parameters of hostꢀguest binding (K, ꢁH, ꢁS, and ꢁG) were extracted from fitting to a single binding site
model (Table 1). When comparing the ITC titrations of the three NARCl receptors (5, 7 and 9) with the guests (11ꢀ13),
several considerations can be made. The ꢁH and ꢁG values indicate the binding process to be exothermic and spontaneous
at 298 K. ꢁH and TꢁS results also indicate that complexation of guests 11ꢀ13 by the receptors to be both enthalpy and
entropy driven in most cases. In three cases (11@5, 13@5, 13@9), negative ꢁS values indicate these process to be
enthalpy driven.
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Figure 4. ITC traces of the titration of guests (10 mM) into host 7 (1 mM) in H2O at 298 K. (a) guest 11, (b) guest 12, (c)
guest 13. All data were fitted into a one siteꢀmodel.
Table 1. Thermodynamic binding parameters of formed complexes between the hosts 5,7 (10 mM) and the guests 9ꢀ11 (1 mM) by ITC[a]
Complex
K (×103)
ꢁH
[kcal/mol]
TꢁS
[kcal/mol]
ꢁG
[kcal/mol]
[Mꢀ1]
11@5
11@7
11@9
12@5
12@7
12@9
13@5
13@7
13@9
0.91±0.09
1.17±0.04
0.95±0.02
2.29±0.30
1.49±0.14
1.05±0.09
1.70±0.50
2.78±0.83
1.37±0.14
ꢀ8.698
ꢀ4.085
ꢀ3.422
ꢀ1.686
ꢀ2.602
ꢀ3.595
ꢀ25.01
ꢀ0.567
ꢀ45.970
ꢀ4.657
0.103
0.644
2.896
1.728
0.527
ꢀ20.58
4.130
ꢀ41.660
ꢀ4.041
ꢀ4.188
ꢀ4.066
ꢀ4.582
ꢀ4.330
ꢀ4.122
ꢀ4.422
ꢀ4.697
ꢀ4.310
[a] ITC in H2O at 298 K.
Complexation Studies via Fluorescence Spectroscopic Analysis. We also used optical titrations to analyze the binding
processes. Fluorescence enhancement and/or quenching were observed from titration experiments conducted at 298 K with
solutions of the hosts 5, 7 and the guests 11ꢀ13. A solution of the guests (0.1 M) at 298 K was titrated into a solution of the
hosts 5 and 7 (2 mL, 125 µM). In all cases, fluorescence spectra were collected after thermal equilibration at 298 K
(Figure 5, Figure S25ꢀS29).
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Figure 5. Fluorescence changes of guest 13 (0.1 M) when titrated to host 7 (125 µM, 2 mL) in H2O at 298 K. Total of 4
additions of guest 13 (0.4, 1.0, 2.0, 8.0 equiv.) were added to host 7.
The red shift in fluorescence emission spectra of the guest 13, registering two maximum (458 nm and 545 nm), was
observed. The lack of a clear isosbestic point indicates that there are more than two species in the system, which is
hypothesized to be the formation of aggregates due to the high concentration of guests used in the system. Fluorescence
enhancement was observed for the guests during the titration with the hosts 5 and 7. The excitedꢀstate vibrational
dynamics of the viologen guests 11ꢀ13 appears to play the key role in the observed enhancement, as well as the red shift of
the emission when the viologens concentration is increased. Studies by Galoppini and coworkers,39 and Pal and
coworkers,40 show similar behavior of a viologen and the dye Brilliant Green, a triphenylmethane derivative, where
restriction of intramolecular bond rotations between the aryl rings induced by complexation/encapsulation with
cucurbiturils resulted in enhanced fluorescence.
CONCLUSION
In conclusion, we synthesized three waterꢀsoluble NARCl receptors (5, 7 and 9) with varying hydrophilicity of the upper
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rim substituents. These receptors exists in the C4v bowlꢀshaped conformation as observed from their relatively simple H
NMR spectra. The binding properties of the NARCl receptors and three waterꢀsoluble viologen derivatives (11ꢀ13) were
investigated in water via NMR, ITC, and fluorescence studies. Binding constants of 103 Mꢀ1 were observed via ITC
analyses. The hosts show higher affinity towards the acetylmethylꢀderived viologen guests 12 and 13 over the methyl
viologen derivative 11. The higher affinity can be attributed to hydrogen bond interactions between the host cationꢀanion
seam and the guest carbonyl groups. This study illustrates the versatility of the NARXs, which in water possesses
hydrophobic cavities and hydrophilic cationꢀanion seam. The ease of functionalization of resorcinarene type receptors into
water soluble NARCl receptors make resorcinarenes a very interesting class of receptor compounds. This versatility
renders the NARXs as suitable receptors for a variety of guests in water.
EXPERIMENTAL SECTION
General Methods. The Nꢀalkyl ammonium resorcinarene chlorides 5, 7 and 9 were synthesized accordingly to
1
modified procedures.26,27,36 1H and 13C NMR spectra were recorded on a Bruker Avance DRX 500 (500 MHz for H and
126 MHz for 13C) and DRX 400 (400 MHz for 1H and 100 MHz for 13C) spectrometers. All signals are given as δ values in
ppm using residual solvent signals as the internal standard. Coupling constants are given in Hz. Melting points were
determined with a Mettler Toledo FP62 capillary melting point apparatus and a Stuart SMP30 melting point apparatus.
Experimental details for the synthesis and characterization data of receptors 5, 6, 7, 8 and 9, and guests 12 and 13 are below.
Compounds 3, and 11 are known compounds and have been reported in previous references.34,35,41 Mass spectrometry
experiments were performed with ABSciex QSTAR Elite ESIꢀQꢀTOF mass spectrometer, equipped with an API 200
TurboIonSpray ESI source from AB Sciex. Nitrogen was used as drying and nebulization gas. VPꢀITC instrument made by
MicroCal were used to determine the molar enthalpy (∆H) of complexation. Subsequent fitting of the data to a 1:1 binding model
using Origin software provides binding constant (K) and the entropy (∆S). Fluorescence spectra were recorded on a PTI
QuantaMasterTM 40 intensity based spectrofluorometer equipped with 814 photomultiplier detection system (V=1000 volts). A
75 W Xenon arc lamp was used as the excitation source. The data for crystals of 11, 11b, and 13 were collected at 123 K for 11b
with an Agilent SuperꢀNova diffractometer using mirrorꢀmonochromatized CuꢀKα (λ=1.54184Å) radiation, and at 100 K for 11
and 13 with the same diffractometer using mirrorꢀmonochromatized MoꢀKα (λ=0.71073Å) radiation.
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General procedure for the synthesis of tetrabenzoxazines from the resorcinarene 3. To a solution of the
resorcinarene 3 (5.5 mmols) and excess formaldehyde (6 mL) in ethanol (40 mL), the amine (23.3 mmols) in ethanol (15
mL) is added slowly and stirred at room temperature for 24 h. The precipitate that separated is filtered, recrystallized in a
methanol/nꢀhexane mixture and dried.
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General procedure for the synthesis of the N-alkyl ammonium resorcinarene chlorides from the
tetrabenzoxazines. Into a solution of the tetrabenzoxazine (0.82 mmol), 3 mL concentrated HCl (37%) and 4 ml H2O in
50 ml isopropanol is heated under reflux. Water and formaldehyde are removed by azeotropic distillation with chloroform.
The remaining isopropanol is evaporated and the crude product triturated with diethyl ether to give the Nꢀalkyl ammonium
resorcinarene chloride.
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NꢀEthanol ammonium resorcinarene chloride (5): CpropanolꢀResorcinarene 3 (4 g, 7.344 mmols), formaldehyde (8 mL),
EtOH (60 mL), 2ꢀaminoethanol (1.86 mL, 30.8 mmols), EtOH (15 ml). The product was a mixture of the sixꢀmembered
and fiveꢀmembered tetrabenzoxazines 4a/4b. This mixture of tetrabenzoxazines was not separated and was used directly in
the next step to obtain the NꢀEthanol ammonium resorcinarene chloride 5. The crude tetrabenzoxazines 4a/4b (1.0 g, 1.129
mmol), 3 ml conc. HCl, 4 ml H2O, 40 ml isopropanol. NꢀEthanol ammonium resorcinarene chloride 5 (0.68 g, 61 %). m.p.
1
> 300 oC; HRMS (ESIꢀTOF) m/z calcd for C52H77N4O16 [5ꢀ4Clꢀ3H]+ 1013.5329, Found 1013.5331, (ꢀ0.2 ppm);. H NMR
(400 MHz, 298K in CD3OD) δ (ppm): 1.53 (m, 8H, CH2), 2.39 (m, 8H, CH2), 3.04 (t, J=5.06 Hz, 8H, OCH2), 3.66 (t,
J=6.20 Hz, 8H, NCH2), 3.78 (t, J=5.12 Hz, 8H, OCH2), 4.32 (s, 8H, ArCH2N), 4.47 (t, J=7.80 Hz, 4H, CH), 7.48 (s, 4H,
ArH); 13C NMR: (100 MHz, 298K in D2O) δ (ppm) = 30.9, 32.0, 35.8, 42.6, 57.6, 62.7, 110.2, 126.7, 128.1, 151.9.
NꢀCyclohexyl Tetrabenzoxazine (6): CpropanolꢀResorcinarene (4 g, 5.5 mmols), formaldehyde (6 mL), ethanol (40 mL),
cyclohexyl amine (2.31 mL, 23.3 mmols), ethanol (15 mL). Tetrabenzoxazine 6 (5.29 g, 79 %). m.p. 218ꢀ220 oC; HRMS
1
(ESIꢀTOF) m/z calcd for C72H101N4O12 [6+H]+ 1213.7411, Found 1213.7422, (ꢀ0.9 ppm);. H NMR (400 MHz, 298 K in
[D6]DMSO) δ (ppm): 0.98ꢀ1.92 (m, 48H, CH2), 2.24 (m, 8H, CH2), 2.43 (m, 8H, CH2), 3.44 (q, J=4.96 Hz, 8H, OCH2),
3.77 (dd, 8H, ArꢀCH2ꢀN), 4.08 (t, J=7.94 Hz, 4H, OCH2), 4.32 (t, J=4.90 Hz, 4H, OH), 5.06 (dd, J=9.64 Hz, 8H, ArꢀCH2ꢀ
O), 7.40 (s, 4H, ArꢀH), 7.62 (s, 4H, ArꢀOH); 13C NMR: (100 MHz, 298 K in [D6]DMSO) δ (ppm) = 24.2, 24.8, 24.9, 25.4,
29.0, 30.3, 31.0, 31.8, 32.0, 42.9, 57.1, 60.4, 60.5, 80.3, 108.6, 122.1, 123.4, 123.8, 148.6.
NꢀCyclohexyl ammonium resorcinarene chloride (7): Tetrabenzoxazine 6 (1.0 g, 0.824 mmol), 3 mL concentrated HCl
o
(37%), 4 ml H2O, 50 ml isopropanol, NꢀCyclohexyl ammonium resorcinarene chloride 7 (1.00 g, 92 %). m.p. > 300 C;
HRMS (ESIꢀTOF) m/z calcd for C68H101N4O12 [7ꢀ4Clꢀ3H]+ 1165.7411, Found 1165.7396, (1.3 ppm). 1H NMR (500 MHz,
298K in D2O) δ (ppm): 0.88 (m, 12H, CH2), 1.11 (m, 12H, CH2), 1.45 (m, 12H, CH2), 1.56 (d, J=13.00 Hz, 8H, CH2), 1.79
(d, J=9.50 Hz, 8H, CH2), 2.26 (m, 8H, CH2), 2.68 (m, 4H, NCH), 3.61 (t, J=6.45 Hz, 8H, OCH2), 4.20 (s, 8H, ArꢀCH2ꢀN),
4.38 (t, J=7.77 Hz, 4H, CH), 7.38 (s, 4H, ArꢀH); 13C NMR: (126 MHz, 298K in D2O) δ (ppm) = 14.0, 23.8, 24.2, 28.7,
29.5, 29.7, 34.2, 38.6, 56.5, 61.5, 65.9, 109.0, 125.3, 126.9, 150.1.
NꢀBenzyl Tetrabenzoxazine (8): CpropanolꢀResorcinarene (5 g, 6.9 mmols), formaldehyde (10 mL), ethanol (60 mL),
benzyl amine (3.18 mL, 29.0 mmols), ethanol (15 mL). Tetrabenzoxazine 8 (4.74 g, 55 %). m.p. 179ꢀ181 oC; HRMS (ESIꢀ
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TOF) m/z calcd for C76H85N4O12 [8+H]+ 1245.6159, Found 1245.6133, (2.1 ppm). H NMR (400 MHz, 298 K in
[D6]DMSO) δ (ppm): 1.36 (m, 8H, CH2), 2.24 (t, J=6.45 Hz, 8H, CH2), 3.42 (m, 8H, CH2), 3.70ꢀ3.85 (m, 16H, ArꢀCH2ꢀN,
OCH2), 4.20 (t, J=7.87 Hz, 4H, OH), 4.33 (t, J=5.42 Hz, 4H, CH), 4.89 (dd, J=9.35 Hz, 8H, ArꢀCH2ꢀO), 7.19ꢀ7.26 (m,
20H, PhꢀH), 7.41 (s, 4H, ArꢀH), 7.64 (s, 4H, ArꢀOH); 13C NMR: (100 MHz, 298 K in [D6]DMSO) δ (ppm) = 29.3, 31.0,
31.1, 45.3, 54.8, 60.5, 107.4, 122.6ꢀ128.5, 138.1, 147.9, 149.6.
NꢀBenzyl ammonium resorcinarene chloride (9): Tetrabenzoxazine 8 (1.0 g, 0.803 mmol), 3 mL concentrated HCl
(37%), 4 ml H2O, 50 ml isopropanol. NꢀBenzyl ammonium resorcinarene chloride 9 (0.95 g, 88 %). m.p. > 300 oC; HRMS
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(ESIꢀTOF) m/z calcd for C72H85N4O12 [9ꢀ4Clꢀ3H]+ 1197.6159, Found 1197.6178, (ꢀ1.6 ppm). H NMR (500 MHz, 298K
in [D6]DMSO) δ (ppm): 1.37 (m, 8H, CH2), 2.32 (m, 8H, CH2), 3.46 (t, J=6.58 Hz, 8H, CH2), 3.98 (br, 8H, NCH2Ph), 4.14
(br, 8H, ArCH2N), 4.31 (t, J=7.57 Hz, 4H, CH), 7.38 (m, 12H, PhꢀH), 7.56 (m, 8H, PhꢀH), 7.64 (m, 4H, ArꢀH), 9.05 (br,
8H, NH2), 9.44 (br, 8H, ArꢀOH); 13C NMR: (126 MHz, 298K in [D6]DMSO) δ (ppm) = 25.8, 29.2, 31.3, 34.5, 41.0, 50.6,
60.7, 109.3, 126.4, 126.8, 128.9, 129.2, 130.4, 131.9, 150.5.
Synthesis of Nꢀmethylcarbonylmethylꢀ4ꢀ,4’ꢀbipyridinium chloride (12). A flask was flame dried and back filled with
nitrogen. 1.72 g (11 mmol) of 4,4’ꢀbipyridyl were added to this and dissolved in 22 mL of dry THF. 3.6 mL (44 mmol) of
chloroacetone were added and the solution was stirred at room temperature overnight. The solvent was removed under
reduced pressure, yielding 2.08 g (76 %) of 12. HRMS (ESIꢀTOF) m/z calcd for C13H13N2O [12+H]+ 213.1022, Found
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213.1020, (1.3 ppm). Hꢀ and 13CꢀNMR spectra were obtained in D2O and [D6]ꢀDMSO. H NMR (400 MHz, 298 K in
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[D6]ꢀDMSO) δ (ppm): 2.34 (s, 3H, CH3), 5.95 (s, 2H, CH2), 8.06 (d, J=6.16, 2H, CH2), 8.71 (d, J=6.88, 2H, ArH), 8.86 (d,
J=6.12 Hz, 2H, ArH), 9.11 (d, J=6.92 Hz, 2H, ArH); 13C NMR: (100 MHz, 298K in [D6]ꢀDMSO) δ (ppm) = 27.6, 68.2,
122.4, 125.4, 146.8, 151.4.
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Synthesis of NꢀmethylꢀN’ꢀmethylcarbonylmethylꢀ4,4’ꢀbipyridinium dihalide (13). 18.10 g (61 mmol) of 11 were
dissolved in 400 mL of acetonitrile. 4.45 mL (61 mmol) of chloroacetone were added and the solution was refluxed
overnight, producing a light brown precipitate. The solid was filtered out and dried under reduced pressure to yield 15.47 g
(73 %) of 11. HRMS (ESIꢀTOF) m/z calcd for C14H16N2O [13ꢀClꢀI]2+ 114.0626 (228.1252 u), Found 114.0629, (3.01
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ppm). Hꢀ and 13CꢀNMR spectra were obtained in D2O and [D6]ꢀDMSO. H NMR (400 MHz, 298 K in [D6]ꢀDMSO) δ
(ppm): 2.36 (s, 3H, CH3), 4.47 (s, 3H, CH3), 5.98 (s, 2H, CH2), 8.82 (d, J=6.80, 2H, ArH), 8.89 (d, J=6.84, 2H, ArH), 9.23
(d, J=6.76 Hz, 2H, ArH), 9.35 (d, J=6.72 Hz, 2H, ArH); 13C NMR: (100 MHz, 298K in [D6]ꢀDMSO) δ (ppm) = 27.6, 48.4,
68.7, 126.6, 147.1, 147.3.
ASSOCIATED CONTENT
Supporting Information
The Supporting Information is available free of charge on the ACS Publications website at http://pubs.acs.org.
Experimental details, copies of the 1H and 13C NMR, NMR and fluorescence spectroscopy, ITC details, and Mass
Spectrometry (PDF).
Xꢀray crystallographic data. CCDC 1525501ꢀ1525503 for 11, 11b and 13, respectively contains the supplementary
crystallographic data for this paper and can be obtained free of charge from the Cambridge Crystallographic Data Centre via
www.ccdc.cam.ac.uk/data_request/cif.
AUTHOR INFORMATION
Corresponding Authors
Eꢀmail: kodiah.beyeh@aalto.fi
Eꢀmail: anslyn@austin.utexas.edu
Eꢀmail: kari.t.rissanen@jyu.fi
ORCID
Ngong Kodiah Beyeh: 0000ꢀ0003ꢀ3935ꢀ1812
Eric Anslyn: 0000ꢀ0002ꢀ5137ꢀ8797
Kari Rissanen: 0000ꢀ0002ꢀ7282ꢀ8419
Author Contributions
The manuscript was written through contributions of all authors.
Notes:
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
The authors gratefully acknowledge financial support from the Academy of Finland (KR: grant nos. 265328, 263256 and 292746;
NKB: grant no. 258653, RHAR: grant no. 272579, EK: grants 284562 and 278743), Aalto University, the University of
Jyvaskyla, Finland, Central China Normal University and National Science Foundation (EVA: grant CHEꢀ1212971), and the
Welch Regents Chair (EVA: Fꢀ0046).
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