Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
S
information for
Shinji Takebayashi, Masato Ikeda, Masayuki Takeuchi* and Seiji Shinkai*
Department of Chemistry and Biochemistry, Graduate School of Engineering,
Kyushu University, Fukuoka 812-8581, Japan, FAX: +81-92-642-3611, e-mail: taketcm@mbox.nc.kyushu-u.ac.jp, seijitcm@mbox.nc.kyushu-u.ac.jp
Experimental.
1 H NMR, absorption spectra, MALDI-TOF MS, ESI-MS spectra and CD spectra were measured with Bruker DMX 600, Shimadzu UV-2500, Perseptive Voyager RP MALDI
TOF spectrompeter, Perseptive Mariner and JASCO J-720 WI, respectively.
Synthesis of 1. 2,2’-Bipyridine-4,4’dicarboxaldehyde (200 mg, 0.94 mmmol) and
2,2’-dipicolylamine (0.34 ml, 1.9 mmol) were dissolved in 30 ml of dichloroethane.
To this solution Sodium triacetoxyborohydride (540 mg, 2.5 mmmol) was added and the resultant mixture was stirred at room temperature for 18 h. After addition of 5 %
NH
4
OH, the insoluble material was filtered off. The filtrate was concentrated in vacuo .
The residual aqueous solution was extracted by chloroform and dried over anhydrous sodium sulfate. The solution was evaporated to dryness, the oil residue was chromatographed (silica gel, chroloform/methanol = 50/1with four drops of 28 % aqueous NH
4
OH) to give yellow solid. The resultant yellow solid was washed by ether to give 1 as white powder (253 mg, 46%). mp 135.9-137.9 °C. 1 H NMR
(600MHz, CDCl
3
, 27 °C,  = ppm, J = Hz) 3.81 (s, 4H), 3.85 (s, 8H), 7.15 (t, J = 6.0,
4H), 7.47 (d, J = 4.8, 2H), 7.59 (d, J = 7.7, 4H), 7.67 (t, J = 7.5, 4H), 8.38 (s, 2H), 8.53
(d, J = 4.5, 4H), 8.63 (d, J = 4.9, 2H); MALDI-TOF MS (dithranol matrix) Calcd.
(found) for [ 1 +H] + . 579.71 (579.71); Calcd for C
36
H
34
N
8
･ 0.5CH
3
OH: C, 73.71; H,
6.10; N, 18.84. Found: C, 73.50; H, 5.93; N, 18.77.

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
Photometric titration of 1 with Zn 2+ or Cu 2+ .
Titration with Zn(ClO
4
)
2
.
Concentration [ 1 ] = 0.4 mM
Solvent
Temperature
Path length
[ Zn(ClO
4
)
2
] = 0 ~ 3.2 mM (0 ~ 8.0 eq.)
Water (buffer : HEPES 25 mM, pH 7.7) / Ethanol = 2 / 1 (v / v)
25 °C
1 mm
1.0
1.0
0.8
0 ~ 1.0 eq.
isosbestic point
293 nm
0.8
2.0 ~ 8.0 eq.
isosbestic point
297.4 nm
0.6
A b s .
0.4
0.6
A b s .
0.4
0.2
0
220 240 260 280 300 320
Wavelength / nm
340
0.2
0
220 240 260 280 300 320
Wavelength / nm
340
1.0
0.8
0.6
A b s .
0.4
0.2
1.0 ~ 2.0 eq.
isosbestic point
296.8 nm
0
220 240 260 280 300 320
Wavelength / nm
340
Fig. S1. Spectrophotometric titration of 1 (0.4 mM) with Zn(ClO
4
)
2
.

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
Titration with Cu(ClO
4
)
2
.
Concentration [ 1 ] = 1.0 mM
Solvent
Temperature
Path length
[ Cu 2+ ] = 0 ~ 6.0 mM (0 ~ 6.0 eq.)
Water (buffer : HEPES 25 mM, pH 7.7) / Ethanol = 2 / 1 (v / v)
25 °C
1 mm
3.0
3.0
2.25 ~ 3.0 eq.
0 ~ 1.25 eq.
isosbestic point
265 nm, 295 nm
2.0
2.0
isosbestic point
300 nm
A b s .
1.0
A b s .
1.0
2.0
A b s .
1.0
0
200 250 300 350
Wavelength / nm
400
3.0
1.25 ~ 2.25 eq.
isosbestic point
272 nm
3.0
2.0
A b s .
1.0
0
200 250 300 350
Wavelength / nm
400
3.25 ~ 6.0 eq.
0
200 250 300 350
Wavelength / nm
400
0
200 250 300 350
Wavelength / nm
Fig. S2. Spectrophotometric titration of 1 (1.0 mM) with Cu(ClO
4
)
2
.
400

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
Characterization of Zn 2+ or Cu 2+ complex of 1 by ESI-MS and 1 H-NMR spectroscopies.
(1) ESI-MS studies.
3.0
+1
N
Zn
N
2+
N
2.5
2.0
6 7 7 . 1
3(ClO
4
-
)
N
N
N
Zn
N
2+
N
1 0 0 5 . 2
1 0 0 7 . 1
1 0 0 9 . 2
1 0 0 3 . 1
1 0 0 8 . 1
1 0 0 6 . 1
1 0 0 4 . 1
1.5
6 4 1 . 3
I n t e n s i t y / 1 0 c p s
1.0
m / z = 1007.0 (calc.)
7 4 1 . 2
0.5
6 5 9 . 2
7 0 1 . 2
9 4 3 . 1
8 4 1 . 2 8 7 9 . 1
9 0 5 . 2
9 2 5 . 1 8 2 3 . 2
8 6 1 . 1
8 0 5 . 1
1 0 0 7 . 1
0
600 700 800 900 m / z
1000 1100 1200 1300
Fig. S3 . ESI-MS spectrum for 1 +2 equiv Zn(ClO
4
)
2
; [ 1 ]=0.4 mM, pH 7.7 in water / ethanol = 2 / 1 (v / v).
8.0
+1
6.0
N
Zn
N
2+
N
N
Zn
N
1 2 6 9 . 0
1 2 7 1 . 1
1 2 7 3 . 1
1 2 7 5 . 1
1 2 6 6 . 9
1 2 7 0 . 1
1 2 7 2 . 1
N N
Zn
2+
5(ClO
4
-
)
1 2 6 4 . 9
1 2 6 5 . 9
1 2 6 8 . 1
4.0
m / z = 1270.8 (calc.) 1 2 7 1 . 1
1 0 0 7 . 1
9 4 3 . 1
I n t e n s i t y / 1 0 c p s
1 2 0 6 . 9
2.0
6 4 1 . 2
6 2 5 . 1
6 0 9 . 0
6 7 7 . 1
7 4 1 . 1
8 8 1 . 1
8 6 1 . 1
9 0 5 . 2
9 2 5 . 1
9 6 5 . 0
1 0 8 9 . 1
1 1 0 7 . 1
1 1 2 2 . 9
1 1 8 8 . 8
1 2 3 0 . 9
0
600 700 800 900 m / z
1000 1100 1200 1300
Fig. S4 . ESI-MS spectrum for 1 +4 equiv Zn(ClO
4
)
2
; [ 1 ]=0.4 mM, pH 7.7 in water / ethanol = 2 / 1 (v / v).

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
1.0
+1
0.8
0.6
N
Zn
N
2+
N
N
Zn
N
N N
Zn
2+
5(ClO
4
-
) m / z = 1270.8 (calc.)
1 2 6 9 . 0
1 2 7 1 . 0
1 2 7 3 . 1
1 2 7 5 . 0
1 2 6 7 . 0
1 2 7 0 . 0
1 2 7 2 . 1 1 2 7 4 . 1
1 2 6 5 . 0
1 2 6 6 . 0
1 2 6 8 . 1
1 2 7 1 . 0
0.4
c p s
1 0 0 7 . 1
1 2 0 6 . 9
0.2
6 0 9 . 0
6 2 5 . 1 9 4 3 . 1
8 2 6 . 0 1 0 8 9 . 1 1 1 4 3 . 1
1 1 8 8 . 8
1 1 6 9 . 1 1 2 3 0 . 9
0
600 700 800 900 1000 1100 1200 1300 m / z
Fig. S5 . ESI-MS spectrum for 1 +8 equiv Zn(ClO
4
)
2
; [ 1 ]=0.4 mM, pH 7.7 in water / ethanol = 2 / 1 (v / v).
2.0
+1
N
Cu
N
2+
N
1.5
N
N
1 0 0 3 . 1
1 0 0 5 . 1
1 0 0 1 . 1
1 0 0 2 . 1
1 0 0 4 . 1
1 0 0 6 . 2
1 0 0 7 . 2
3(ClO
4
-
)
N
Cu
N
2+
N m / z = 1003.0 (calc.)
1.0
I n t e n s i t y / 1 0 c p s
8 6 5 . 2
9 3 9 . 2
0.5
8 7 5 . 0
8 2 1 . 1
6 4 8 . 1
1 0 0 3 . 1
6 7 6 . 1
7 3 9 . 1
7 5 7 . 2
8 0 3 . 1
8 3 8 . 9
9 0 1 . 1
0
600 700 800 900 m / z
1000 1100 1200 1300
Fig. S6 . ESI-MS spectrum for 1 +2 equiv Cu(ClO
4
)
2
; [ 1 ]=0.4 mM, pH 7.7 in water / ethanol = 2 / 1 (v / v).

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
5.0
+1
4.0
3.0
N
Cu
N
2+
N
N
Cu
N
N N
Cu
2+
5(ClO
4
-
) m / z = 1265.8 (calc.)
1 2 6 4 . 0
1 2 6 6 . 0
1 2 6 8 . 1
1 2 6 5 . 1
1 2 6 7 . 0
1 2 7 0 . 0
1 2 6 8 . 8
1 2 7 1 . 1
1 1 2 8 . 0
2.0
c p s
9 3 9 . 2
1.0
6 4 8 . 1
6 9 5 . 5
8 7 5 . 0
8 6 5 . 2 9 0 1 . 1
8 3 8 . 9
1 0 0 3 . 1
1 0 6 4 . 1
1 0 9 7 . 9
1 0 8 4 . 0
1 1 3 8 . 1
1 1 6 4 . 1
1 2 0 1 . 9
1 2 6 6 . 0
0
600 700 800 900 m / z
1000 1100 1200 1300
Fig. S7 . ESI-MS spectrum for 1 +3 equiv Cu(ClO
4
)
2
; [ 1 ]=0.4 mM, pH 7.7 in water / ethanol = 2 / 1 (v / v).
6.0
+1
5.0
N
Cu
N
2+
N
N
Cu
N
1 2 6 3 . 9
1 2 6 6 . 1
1 2 6 8 . 1
1 2 6 2 . 1
1 2 6 3 . 0
1 2 6 5 . 1
1 2 6 7 . 1
1 2 7 0 . 1
1 2 6 9 . 0
1 2 7 1 . 2
4.0
N N
Cu
2+
5(ClO
4
-
)
3.0
m / z = 1265.8 (calc.)
1 2 0 2 . 0
I n t e n s i t y / 1 0 c p s
2.0
1 2 6 6 . 1
1 1 6 4 . 0
7 5 8 . 0
7 4 7 . 3 7 7 9 . 1
8 3 8 . 9
8 3 8 . 9
9 3 9 . 2
1.0
6 4 8 . 0
6 2 1 . 1
1 0 0 3 . 1
1 1 0 0 . 0
1 0 8 2 . 0
1 0 6 4 . 0
0
600 700 800 900 m / z
1000 1100 1200 1300
Fig. S7 . ESI-MS spectrum for 1 +3.5 equiv Cu(ClO
4
)
2
; [ 1 ]=0.4 mM, pH 7.7 in water / ethanol = 2 / 1 (v / v).

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
(2) 1 H-NMR studies.
N
N
N
N i
N h f g
N e d c
N
N b a
Zn
2+
0 eq.
h
8.59
a
8.40
i cd
7.75
8.20
7.82
g
7.58
b
7.30
Zn
2+
2 eq.
Zn
2+
8 eq.
Fig. S8 . ppm
9.25
9.00
8.75
8.50
8.25
8.00
7.75
7.50
7.25
7.00
6.75
6.50
1 H-NMR spectra for 1 + Zn(ClO
4
)
2
; [ 1 ]=1.0 mM, pD 7.7 in D
2
O / ethanold
6
= 2 / 1 (v / v) at 5 °C.

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
Kinetic Studies.
To a solution of 1 in ethanol/water (HEPES, 16 mM) = 1:2 v/v solution at pH 7.7 and 25
˚C was added freshly prepared stock solution of Zn(ClO
4
)
2
or Cu(ClO
4
)
2
. The change in the absorbance at 406 nm was monitored with Shimadzu UV-2500 just after the addition of HPNP. The pseudo first rate constants were determined from the initial values of at least three measurements (< 5 % conversion).
Control experiments using compounds 2-4.
2.5
2.0
1.5
• ( Z n 2 +
• ( Z n 2 +
1.0
o b s
/ 1 0
- 4
s
- 1
0.5
• Z n
+
2 +
• ( Z n 2 +
3
•(Zn 2+
)
2
0
(a) (b) (c) (d)
Figure S9.
Pseudo-first-order rate constants of the hydrolysis of HPNP (0.8 mM) in
33 % ethanol/water (HEPES, 25 mM), pH 7.7 at 25 °C; (a) [ 1 ] = 0.4 mM, [Zn(ClO
4
)
2
] =
2.0 mM, (b) [ 2 ] = 0.4 mM, [Zn(ClO
4
)
2
] = 2.0 mM (c) [ 2 ] = [ 4 ] = 0.4 mM, [Zn(ClO
4
)
2
] =
2.0 mM, (d) [ 3 ] = 0.4 mM, [Zn(ClO
4
)
2
] = 2.0 mM.
1.5
1.0
• ( C u 2 + • ( C u 2 +
• C u 2 +
• ( C u 2 +
+
• ( C u 2 +
/ 1 0
- 3
s
- 1
0
(e) (f) (g) (h)
Figure S10.
Pseudo-first-order rate constants of the hydrolysis of HPNP (1.0 mM) in
33 % ethanol/water (HEPES, 25 mM), pH 7.7 at 25 °C; (e) [ 1 ] = 1.0 mM, [Cu(ClO
4
)
2
] =
3.0 mM, (f) [ 2 ] = 1.0 mM, [Cu(ClO
4
)
2
] = 2.0 mM (g) [ 2 ] = [ 4 ] = 1.0 mM, [Cu(ClO
4
)
2
] =
3.0 mM, (h) [ 3 ] = 1.0 mM, [Zn(ClO
4
)
2
] = 2.0 mM.

Supplementary Material for Chemical Communications
This journal is © The Royal Society of Chemistry 2004
Lineweaver-Burk plots for HPNP cleavage.
2.5
2.0
(a)
1.5
1.0
0.5
(b)
0
0 0.5
1.0
[HPNP]
-1
1.5
/ 10
3
2.0
2.5
M
-1
Figure S11 . Lineweaver-Burk plots for HPNP cleavage; in 33 % ethanol/water
(HEPES, 25 mM), pH 7.7 at 25 °C; (a) [ 1 ] = 0.4 mM, [Zn(ClO
4
)
2
] = 0.8 mM, (b) [ 1 ] =
0.4 mM, [Zn(ClO
4
)
2
] = 2.0 mM.
4.0
3.0
2.0
(a)
- 1
/ 1 0 M
- 1
(b)
0
0 1.0
2.0
3.0
4.0
[HPNP]
-1
/ 10
3
M
-1
5.0
Figure S12 . Lineweaver-Burk plots for HPNP cleavage; in 33 % ethanol/water
(HEPES, 25 mM), pH 7.7 at 25 °C; (a) [ 1 ] = 1.0 mM, [Cu(ClO
4
)
2
] = 2.0 mM, (b) [ 1 ] =
1.0 mM, [Cu(ClO
4
)
2
] = 3.0 mM.