Supporting Information
NUCLEOPHILIC REACTIVITY OF BIOTHIOLS TOWARD COUMARIN-BASED
DERIVATIVES CONTAINING A CHALCONE MOIETY
Olimpo García-Beltrán,1,2 Christopher González,3 Edwin G. Pérez,3 Bruce K. Cassels,1,4 José G.
Santos,3 Daniela Millán,3 Natalia Mena,5 Paulina Pavez3* and Margarita E. Aliaga.3*
1
Department of Chemistry, Faculty of Sciences, University of Chile, Santiago, Chile.
2
Departamento de Ciencias Químicas, Facultad de Ciencias Exactas,
Universidad Andrés Bello, Avenida República 275, Piso 3, Santiago. Chile
3
Facultad de Química, Pontificia Universidad Católica de Chile, Av. Vicuña Mackenna 4860,
Santiago 6094411, Chile
4Institute
5
for Cell Dynamics and Biotechnology, University of Chile, Santiago, Chile.
Department of Biology, Faculty of Sciences, University of Chile, Santiago, Chile.
* Corresponding authors. Tel.: 56-2 3544743; Fax: 56-2 3544744; e-mail addresses:
ppavezg@uc.cl; mealiaga@uc.cl
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Table of contents
Synthesis and characterization.
Figure S1. (A) 1H NMR spectrum of ChC2 and (B) 13C NMR spectrum of ChC2.
Figure S2. (A) 1H NMR spectrum of ChC3 and (B) 13C NMR spectrum of ChC3.
Kinetic studies.
Figure S3. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with Cys. () pH 7.4
and (●) pH 6.9.
Figure S4. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with Cys-Gly. () pH
7.4 and (●) pH 6.9.
Figure S5. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with GSH. () pH 7.4
and (●) pH 6.9.
Figure S6. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with Hcy. () pH 7.4
and (●) pH 6.9.
Figure S7. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with Cys. () pH 7.4
and (●) pH 6.9.
Figure S8. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with Cys-Gly. () pH
7.4 and (●) pH 6.9.
Figure S9. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with GSH. () pH 7.4
and (●) pH 6.9.
Figure S10. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with Hcy. () pH 7.4
and (●) pH 6.9.
S2
Synthesis and characterization.
O
O
OH
c
b
HO
1
Scheme
R5
R4
O a
HO
O
O
O
MeO
O
2
O
MeO
3
R1
R2
R3
R4
R5
ChC1
H
H
H
H
H
ChC2
OMe
H
H
H
H
OMe
H
H
H
ChC3
H
O
O R1
ChC
R3
R2
1. Synthetic route to probes ChC2 -ChC3. Reagents and conditions: Resorcinol was formylated with POCl3, DMF,
acetonitrile, 0-5 °C, 2h; a) CH3COCH2COOC2H5, ethanol, reflux, 6 h; b) Me2SO4, K2CO3, DMF, H2O, 1 h c) 2methoxybenzaldehyde (ChC2), 3-methoxybenzaldehyde (ChC3), piperidine, CH2Cl2, reflux, 12 h.
Resorcinol was formylated (Vilsmeier-Haack) giving 2,4-dihydroxybenzaldehyde (1),[1] which was
subsequently condensed with ethyl acetoacetate (Henry-Knoevenagel) and cyclized in a single step
to afford 3-acetyl-7-hydroxycoumarin (2).
[´2]
The 7-hydroxycoumarin obtained was methylated
with methyl sulfate (3),4 and finally ChC2 and ChC3 were prepared by appropriate adaptations of a
literature procedure.[3]
The synthesis of 3-cinnamoyl-7-methoxy-2H-chromen-2-one (ChC1), was recently reported by
us, [4] and analogous synthetic procedures were used for the preparation of ChC2 and ChC3.
(E)-7-Methoxy-3-(3-(2-methoxyphenyl)acryloyl)-2H-chromen-2-one (ChC2).
S3
H
H
5
H
6
MeO 7
H
8
H
H
3' 6'' 5'' H
3
2' 1''
4''
H
O 2 O MeO 2'' 3'' H
1
H
4
O
1'
From 3-acetyl-7-methoxy-2H-chromen-2-one (0.44 g, 2 mmol) and o-methoxybenzaldehyde (0.27
g, 2 mmol), 0.330 g yellow solid, 49.1 %. m.p. 184-186 °C; 1H NMR (CDCl3): δ 8.56 (s, 1H, =CH), 8.21 (d, 1H, Ar-CH=, J = 15.8 Hz), 8.04 (d, 1H, CO-CH=, J = 15.8 Hz), 7.71 (d, 1H, Ar-H, J =
7.6 Hz), 7.56 (d, 1H, Ar-H, J = 8.4 Hz), 7.37 (t, 1H, Ar-H, J = 7.9 Hz), 6.98 (t, 1H, Ar-H, J = 7.7
Hz), 6.92 (d, 1H, Ar-H, J = 8.2 Hz), 6.90 (d, 1H, Ar-H, J = 8.5 Hz), 6.85 (s, 1H, Ar-H), 3.91 (s, 6H,
2 OCH3);
13
C NMR (DMSO-d6): δ 55.6, 56.0, 100.4, 111.2, 112.5, 113.7, 120.8, 121.9, 124.1,
124.6, 129.3, 131.3, 132.0, 139.9, 148.2, 157.6, 159.0, 159.8, 165.0, 186.8.
Figure S1. (A) 1H NMR spectrum of ChC2.
S4
Figure S1. (B) 13C NMR spectrum of ChC2.
(E)-7-Methoxy-3-(3-(3-methoxyphenyl)acryloyl)-2H-chromen-2-one (ChC3)
H
H
O
H
H
H
H
MeO
O
O
H
H
H
H
OMe
From 3-acetyl-7-methoxy-2H-chromen-2-one (0.44 g, 2 mmol) and m-methoxybenzaldehyde (0.27
g 2 mmol), 0.280 g yellow solid, 41.7 %, m.p. 164-166 °C; 1H NMR (CDCl3): δ 8.49 (s, 1H, =CH), 7.90 (d, 1H, Ar-CH=, J = 15.9 Hz), 7.73 (d, 1H, CO-CH=, J = 15.9 Hz), 7.48 (d, 1H, Ar-H, J =
8.8 Hz), 7.22 (d, 1H, Ar-H, J = 7.8 Hz), 7.18 (d, 1H, Ar-H, J = 7.3 Hz), 7.09 (s, 1H, Ar-H), 6.87
(dd, 1H, Ar-H, J = 8.8, 2.0 Hz), 6.22 (dd, 1H, Ar-H, J = 7.8, 2.0 Hz), 3.83 (s, 3H, OCH3), 3.76 (s,
3H, OCH3); 13C NMR (CDCl3): δ 55.8, 56.5, 100.8, 112.8, 113.9, 114.3, 117.1, 121.7, 122.0, 124.9,
130.3, 131.8, 136.8, 144.8, 149.0, 158.1, 160.2, 160.3, 165.8, 186.8.
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Figure S2. (A) 1H NMR spectrum of ChC3.
S6
Figure S2. (B) 13C NMR spectrum of ChC3.
[1]. W. Meldenson, S. Hayden, Synth. Commun. 1996, 26, 603-610.
[2]. Y-F. Sun, Y.-P. Cui, Dyes Pigments 2008, 78, 65-76.
[3]. H. F Lewis, S. Shaffer, W. Trieschmann, H. Cogan, Ind. Eng. Chem. 1930, 22, 34-36.
[4]. O. García-Beltrán, N. Mena, E. G. Pérez, B. K. Cassels, M. T. Nuñez, F. Werlinger, D.
Zavala, M. E. Aliaga, P. Pavez, Tetrahedron Lett. 2011, 52, 6606-6609.
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0.05
kobsd / s
-1
0.04
0.03
0.02
0.0000
0.0001
0.0002
0.0003
0.0004
-
[RS ]/M
Figure S3. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with Cys. () pH 7.4
and (●) pH 6.9.
0.020
kobsd / s
-1
0.015
0.010
0.005
0.0000
0.0005
0.0010
0.0015
-
[RS ]/M
Figure S4. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with Cys-Gly. ()
pH 7.4 and (●) pH 6.9.
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0.020
kobsd / s
-1
0.018
0.016
0.014
0.00000
0.00005
0.00010
-
[RS ]/M
Figure S5. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with GSH. () pH
7.4 and (●) pH 6.9.
0.016
kobsd / s
-1
0.014
0.012
0.010
0.0000
0.0001
0.0002
0.0003
-
[RS ]/M
Figure S6. Plots of kobsd vs thiolate ion concentration for the reaction of ChC2 with Hcy. () pH
7.4 and (●) pH 6.9.
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0.015
0.012
kobsd / s
-1
0.009
0.006
0.003
0.000
0.0000
0.0001
0.0002
0.0003
0.0004
-
[RS ]/M
Figure S7. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with Cys. () pH 7.4
and (●) pH 6.9.
0.004
kobsd / s
-1
0.006
0.002
0.000
0.001
0.002
-
[RS ]/M
Figure S8. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with Cys-Gly. ()
pH 7.4 and (●) pH 6.9.
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kobsd / s
-1
0.003
0.002
0.001
0.00000
0.00005
0.00010
-
[RS ]/M
Figure S9. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with GSH. () pH
7.4 and (●) pH 6.9.
0,0035
kobsd / s
-1
0,0030
0,0025
0,0020
0,0000
0,0001
0,0002
0,0003
-
[RS ]/M
Figure S10. Plots of kobsd vs thiolate ion concentration for the reaction of ChC3 with Hcy. () pH
7.4 and (●) pH 6.9.
S11