Data in Figure 5E showed that the complex band of biotin labeled EGFR nucleotid

Cells were grown on 75 cm2 cell culture with EMEM medium for breast carcinoma and supplemented with 10% fetal bovine serum, and 1% penicillin streptomycin solution in a humidified atmosphere of 5% CO2, 95% air at 37 C. Cell proliferation assay Cell proliferation assay of compounds was evaluated in BT 20 cells, and was compared with Amuvatinib PDGFR 阻害剤 that of doxorubicin. Cell proliferation assay was carried out using CellTiter 96 aqueous one solution cell proliferation assay kit. Briefly, upon reaching about 75 80% confluence, 5000 cells well were plated in 96 well microplate in 100 uL media. After seeding for 72 h, the cells were treated with 50 uM compound in triplicate. DOX was used as the positive control. Incubation was carried out at 37 C in an incubator supplied with 5% CO2 for 72 h.<br><br> At the end of the sample exposure period, 20 uL CellTiter 96 aqueous AT-406 solution was added. The plate was returned to the incubator for 1 h in a humidified atmosphere at 37 C. The absorbance of the formazan product was measured at 490 nm using microplate reader. The blank control was recorded by measuring the absorbance at 490 nm with wells containing medium mixed with CellTiter 96 aqueous solu tion but no cells. Results were expressed as the percentage of the control. The percentage of cell survival was calculated as × 100%. Results and discussion Chemistry Naphthopyran analogs described here were synthesized following the synthetic routes outlined in Figure 3. The compounds were synthesized according to the previously reported general procedure by a one pot reaction through the condensation of a substituted aromatic aldehyde, malonitrile, and or B naphtol in ethanol and water in the presence of diam monium hydrogen phosphate.<br><br> Src kinase inhibitory activity AG-490 EGFR 阻害剤 The results of Src kinase inhibitory activity of compounds 4a n are shown in Table 1. Compounds 4a, 4d, 4i, 4m, and 4n exhibited higher Src kinase inhibitory activity when compared with that of other compounds. Unsubstituted phenyl analog 4a showed an IC50 value of 28. 1 uM and was the most potent compound in this series. 2 Chlorophenyl substituted analog 4d was one of the potent compounds in naphthol series with an IC50 value of 34. 7 uM. Similarly compound 4j with 3 chlorophenyl substitution was a moderately effective com pound in B naphthol series with an IC50 value of 41. 7 uM.<br><br> In addition, 2,3 dichlorophenyl substituted compounds 4f and 4l were among potent inhibitors in both and B naphthol series of compounds. However, substitution of 2,6 dichlorophenyl in compound 4g led to loss of inhibitory activity, suggesting that chloro substitution at positions 2 and 6 of phenyl ring is detrimental in Src kinase inhibition. Compounds 4m and 4n with 3 hydroxyphenyl and 4 methoxyphenyl moieties, respectively, were among potent compounds in B naphthol series. 3 Nitrophenyl substitu tion resulted in loss of activity in compounds 4e and 4k in both and B naphthol series of compounds. Similarly compounds 4b and 4c did not show any significant inhibitory activity against Src kinase. On the other hand, compounds 4h with pyridine ring sub stitution and compound 4i with 1 methylnitroimidazole moiety showed good inhibitory activity against Src kinase compared to other phenyl substituted compounds.