, 2012). Here, we study the mechanism of ATZD’s selective cytotoxicity (AC-4, AC-7, AC-10 and AC-23) in human colon carcinoma HCT-8 Akt activation cells. The chemical data and synthetic procedures for (5Z)-5-acridin-9-ylmethylene-3-(4-methylbenzyl)-thiazolidine-2,4-dione (AC-4), (5ZE)-5-acridin-9-ylmethylene-3-(4-bromo-benzyl)-thiazolidine-2,4-dione (AC-7), (5Z)-5-(acridin-9-ylmethylene)-3-(4-chloro-benzyl)-1,3-thiazolidine-2,4-dione (AC-10) and (5ZE)-5-(acridin-9-ylmethylene)-3-(4-fluoro-benzyl)-1,3-thiazolidine-2,4-dione

(AC-23) are reported elsewhere ( Barros et al., 2012, Mourão et al., 2005 and Silva et al., 2001). Thiazolidine-2,4-dione was N-(3)-alkylated in the presence of potassium hydroxide, which enabled the thiazolidine potassium salt to react with the substituted benzylhalide in a hot alcohol medium. The thiazacridine derivatives were synthesised by the nucleophilic addition of substituted Epacadostat 3-benzyl-thiazolidine-diones on 3-acridin-9-yl-2-cyano-acrylic acid ethyl ester. The mechanisms of cytotoxic action for the thiazacridine derivatives were studied as single

Z isomers for AC-4 and AC-10. The AC-7 and AC-23 compounds were studied as isomeric mixtures, but the Z isomer was the major stereoisomer. The Saccharomyces cerevisiae strains in this study were acquired from Euroscarf (European Saccharomyces cerevisiae Archive for Functional Analysis). The following S. cerevisiae genotypes were used in this study: BY-4741 (MATa; his3Δ 1; leu2Δ 0; met15Δ 0; ura3Δ 0); Top1Δ (YOL006c), same as BY4741 with YOL006c::kanMX4; Top3Δ (YLR234w), same as BY4741 with YLR234w::kanMX4. The media, solutions and buffers were prepared as previously described ( Burke et al., 2000). Complete medium (YPD), containing 1% yeast extract, 2% peptone and 2% glucose was used for routine growth. The stationary-phase cultures were obtained by inoculating an isolated colony into liquid YPD medium and incubating the culture at 28 °C for 72 h with shaking (for aeration). Cultures in the exponential phase were obtained by inoculating 5 × 106 cells/ml of the stationary-phase YPD culture into fresh YPD medium at 28 °C for 2 h. The cell concentrations were

determined in a Neubauer chamber using HSP90 a light microscope (LO, Laboroptik GmbH, Bad Homburg, Hessen, Germany). The cytotoxicity of ATZD was evaluated using human colon carcinoma HCT-8 cells donated by the Children’s Mercy Hospital, Kansas City, MO, USA. The cells were maintained in RPMI-1640 medium supplemented with 10% foetal bovine serum, 2 mM glutamine, 100 μg/ml streptomycin and 100 U/ml penicillin. The cells were kept in tissue-culture flasks at 37 °C in a humidified atmosphere with 5% CO2 and were harvested with a 0.15% trypsin–0.08% EDTA, phosphate-buffered saline solution (PBS). The following experiments were performed to determine ATZD’s cytotoxic mechanisms in HCT-8 cells. For all cell-based assays, the HCT-8 cells were seeded (0.