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(Table 1). Their IC50s showed that all these three compounds have
less activity in all these cell lines, indicating that the activity was
diminished while the indazole ring was replaced by the azaindaz-
ole ring. These data suggested that the indazole ring is an impor-
tant moiety for anticancer activity and the trifluoromenthy or
halogen substituent at the 4 position of indazole ring is favorable.
We evaluated the acute toxicity for the representative com-
pounds 5a, 5b and 5i in mice. The highest single oral dose of
2000 mg/kg for these compounds was tolerated without any sign
of toxicity, while the tolerated single oral dose of sorafenib was
1460 mg/kg in mice. These results suggested that these new syn-
thesized diarylurea compounds might have lower toxicity than
sorafenib. Further toxicities of these compounds are currently
underway.
In conclusion, a new series of indazole based diarylurea com-
pounds were synthesized and their activity against human tumor
cell lines was examined. Among them, compounds with trifluoro-
menthy and halogen substituents showed better effect than sorafe-
nib. Compounds 5a, 5b and 5i were found to have lower toxicity
than sorafenib. Compound 5i could be developed as a promising
agent for treatment of cancers.
15. Mir, O.; Coriat, R.; Blanchet, B.; Durand, J. P.; Boudou-Rouquette, P.; Michels, J.;
Ropert, S.; Vidal, M.; Pol, S.; Chaussade, S.; Goldwasser, F. PLoS One 2012, 7,
37563.
16. Villanueva, A.; Llovet, J. M. Clin. Cancer Res. 1824, 2012, 18.
17. NCI-H460, A549, HT-29, Lovo, HepG2, Bel-7402, OS-RC-2, SGC-7901 and MDA-
MB-231 cells were purchased from the American Type Cell Culture Collection
(ATCC, USA) and maintained in RPMI-1640 or DMEM medium supplemented
with 10% (v/v) heat-inactivated fetal bovine serum, penicillin–streptomycin
(100 IU/mL–100 lg/mL), 2 mM glutamine, and 10 mM HEPES buffer at 37 °C in
a humidified atmosphere (5% CO2–95% air). Culture medium was changed
every 2 days. Cells were harvested by brief incubation in 0.02% (w/v) trypsin in
PBS (ICN, Aurora, USA), seeded in 96-well plates (3 Â 103 per well) and then
were incubated in humidified atmosphere for 24 h prior to exposure to various
concentrations of test compounds for 72 h. Cell viability was assessed using the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay
Acknowledgment
This project was supported by Jinan ‘5150’ Plan to Dr. Wenbao
Li.
via adding 20 lL of MTT (5 mg/mL, Sigma, USA) and incubating for 4 h. The
absorbance at 570 nm was measured on a microplate reader (Perkin-Elmer,
USA), and the IC50s were calculated.
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