D. Rodriguez-Lucena et al. / Tetrahedron 67 (2011) 2149e2154
2153
a 5% solution of TFA in CH2Cl2 (2 mL). The solution was stirred for
3 h at 20 ꢁC then solvents were evaporated under reduced pressure.
The oily residue was dissolved in water (5 mL) and freeze-dried,
leading to compound 12 (61 mg, 99%) isolated as a white bis-TFA
salt. Rf¼0.55 (4:1:1 n-BuOH/H2O/AcOH); 1H NMR (300 MHz,
Substances Naturelles at Gif-sur-Yvette (France). The human cell
lines KB (mouth epidermoid carcinoma) and HepG2 (hep-
atocarcinoma) were obtained from ECACC (Salisbury, UK) and
grown in DMEM medium supplemented with 10% fetal calf serum
(FCS), in the presence of penicillin, streptomycin, and fungizone,
in a 75 cm2 flask, under an atmosphere containing 5% CO2. By
contrast, HCT116, HT29, and HCT15 (colon adenocarcinoma),
MCF7 (breast adenocarcinoma), MCF7R (MCF7 cell lines resistant
to doxorubicine), SK-OV-3 (ovary adenocarcinoma from NCI), PC-3
(prostate adenocarcinoma), A549 (lung carcinoma), HL60 (pro-
myelocytic leukemia), K562 (chronic myelogenous leukemia), and
SF268 (glioblastoma from NCI) cells were grown in RPMI medium.
DMSO-d6)
d 12.24 (br s, 2H), 12.21 (br s, 2H), 8.79 (br t, 2H,
J¼5.7 Hz), 8.95 (d, 2H, J¼2.4 Hz), 9.06e8.82 (m, 2H), 8.55 (s, 2H),
7.93 (dd, 2H, J¼8.7 Hz), 7.14 (d, 2H), 4.21e4.08 (m, 4H), 3.47e3.38
(m, 4H), 3.33 (s, 4H); 13C NMR (75 MHz, DMSO-d6)
d 187.7, 167.0,
161.6, 159.1, 152.6, 132.7, 129.9, 128.0, 122.1, 119.1, 116.5, 45.2, 42.9,
41.2; ESI-MS m/z 673.1 ([MþH]þ). High resolution ESI-MS: m/z
found 673.1025 ([MþH]þ); m/z calcd for C28H29N6O6S4: 673.1026.
Cells were plated in 200
microplates and were treated 24 h later with compounds dis-
solved in DMSO at concentrations of 1 and 10 M, using a Biomek
mL of medium, in 96-well tissue culture
4.2.7. Benzoic acid, 3,30-[1,2-ethanediylbis(imino-2,1-ethanediylimi-
nocarbonyl-4,2-thiazolediyl)]-bis[4-hydroxy],1,10-dimethyl ester,2,2,2-
trifluoroacetate (1:2) (13). The compound 7 (113 mg, 0.13 mmol) was
dissolved in a solution of 5% TFA in CH2Cl2 (2 mL). The solution was
stirred for 5 h at 20 ꢁC before being evaporated under reduced
pressure. The oily residue was dissolved in water (5 mL) and freeze-
dried to afford the compound 13 (93 mg, 80% yield) isolated as
m
3000 (Beckman-Coulter). Controls received the same volume of
DMSO (1% final volume). After 72 h of incubation, MTS reagent
(Promega) was added and the plates incubated for 3 h at 37 ꢁC.
Absorbance was monitored at 490 nm and the results are
expressed as the inhibition of cell proliferation, calculated as the
ratio [(1ꢀ(OD490 treated/OD490 control))ꢂ100] in triplicate
experiments.
1
a white bis-TFA salt. Rf¼0.49 (10:1:1 CH3CN/H2O/NH4OH); H NMR
(300 MHz, CD3OD)
d
8.65 (d, 2H, J¼2.4 Hz), 8.23 (s, 2H), 7.89 (dd, 2H,
J¼8.7 Hz), 6.97 (d, 2H), 3.89 (s, 6H), 3.84e3.80 (m, 4H), 3.65 (s, 4H),
3.48e3.44 (m, 4H); 13C NMR (75 MHz, CD3OD)
168.1, 165.5, 165.3,
160.7, 149.0, 133.8, 131.2, 126.4, 123.0, 120.0, 117.6, 52.7, 49.9, 44.9,
d
Acknowledgements
37.4; 19F NMR (188 MHz, CD3OD)
d
ꢀ73.4 (TFA salt); ESI-MS m/z
669.1 ([MþH]þ), 667.0 ([MꢀH]ꢀ). High resolution ESI-MS: m/z found
669.1805 ([MþH]þ); m/z calcd for C30H33N6O8S2: 669.1796.
G.L.A.M., D.R.L., and I.J.S. would like to thank the Centre National
de la Recherche Scientifique (CNRS) and the Agence Nationale pour
la Recherche (ANR-05-JCJC-0181-01) for global financial support.
4.2.8. Benzoic acid, 3,30-[1,2-ethanediylbis(imino-2,1-ethanediylimino-
carbonothioyl-4,2-thiazolediyl)]-bis[4-hydroxy],1,10-dimethyl ester,2,2,2-
ꢀ
Authors would like to thank Region Alsace for financial support and
for providing a postdoctoral fellowship to D.R.L.
trifluoroacetate (1:2) (14). The compound 8 (68 mg, 75 mmol) was
dissolved in a 5% solution of TFA in CH2Cl2 (2 mL). The solution was
stirred for 6 h at 20 ꢁC and solvents were subsequently evaporated
under reduced pressure. The oily residue was dissolved in water
(5 mL) before being freeze-dried to afford the compound 14 (63 mg,
90%) isolated as a white bis-TFA salt. Rf¼0.48 (10:1:1 CH3CN/H2O/
References and notes
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NH4OH); 1H NMR (300 MHz, CD3OD)
d
8.89 (d, 2H, J¼2.4 Hz), 8.50
(s, 2H), 7.94 (dd, 2H, J¼8.7 Hz), 7.04 (d, 2H), 4.29 (br t, 4H, J¼5.7 Hz),
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The antiproliferative activity of 13 cancer cell lines was eval-
ꢁ
uated at the Ciblotheque Cellulaire de l’Institut de Chimie des