New Glutaric The Ketene Class And Quinazoline Compounds In Vitro Anti-tumor Biological Activity And Mechanism Of A Preliminary Study

Quinazoline derivatives exhibit antitumor,antivirus,insecticidal,antidiabetic,acaricide and anti-bacteria biological activities.For example,Fluquinconazole as anti-bacterial agent, Fenazaquin as acaricide agent,ZD 1839 as antitumor agent and Pyrazosin as high blood pressure treatment agent and so on.ZD1839 is an inhibitor of epidermis growth factor receptor(EGFR) tyrosine kinase,4-amino quinazoline derivatives have inhibition activity on EGFR.Curcumin has broad biological activities including anti-inflammatory,anti-oxidant, anti-bacteria,anti-HIV,anti-angiogenesis and anti-cancer activities,especially the anti-angiogenesis and anti-cancer properties.Nowadays,the design and synthesis of pentadienone compounds with high biological activities have attracted more and more attention.In order to investigate the structure-activity relationship of pentadienone and quinazoline derivatives and their antitumor mechanism,more than 200 new pentadienone and quinazoline derivatives synthesized in Center for Research and Development of Fine Chemicals of Guizhou University were screened against androgen-independent prostate cancer cell line PC 3 and fibroblast cell line NIH 3T3 cell through MTT colorimetric assay.Furthermore,pentadienone derivative 2005244(1E,4E)-1,5-bis(2-hydroxyphenyl)penta-1,4-dien-3-one and quinazoline derivative 2007238 N-(4-chlorophenyl)-5,6,7-trimethoxyquinazolin-4-amine dihydrochloride were used to investigate the anti-tumor mechanism on PC 3 cells.The study included:(1) To evaluate inhibition effect of the compounds on PC 3 cell growth by MTT colorimetric assay;(2) Morphologic changes of PC 3 cell and NIH 3T3 cell were observed through light microscope and the cytotoxicity of the compounds to PC 3 cell and NIH 3T3 cell were assessed;(3) The membrane damage effect of the compounds on PC 3 cell was investigated by LDH release analysis;(4) To assess the cytotoxieity of the compounds to PC 3 cell by Trypan Blue staining;(5) To observe morphologic changes of PC 3 cell and assess the cytotoxicity and apoptosis-inducing effect of the compounds to PC 3 cell by AO/EB staining;(6) To evaluate apoptosis-inducing effect of the compounds on PC 3 cell by agrose gel electrophoresis of DNA;(7) To determine the effects of the compounds on total protein in PC 3 cell by SDS-PAGE;(8) To determine the inhibition effect of the compounds on the phosphorylation of Erk 1/2 and CDK4 by Western Blot analysis.It was found the antitumor activity of pentadienone compound 2005244 was due to its cytotoxicity in a dose and time dependent manner.Its in vitro IC₅₀ values of 6,12,24,36,48,72 h for PC 3 in vitro were 31.8,14.9,11.5,11.4,9.0 and 3.9μM respectively.It was found that the cytotoxicity of quinazoline compound 2007238 was very low.Its IC₅₀ value of 72 h for PC 3 cell was 24.5μM.Also,it inhibited the phosphorylation of Erk1/2 at 10μM.It suggested that 2007238 may inhibit PC 3 cell proliferation through Erk1/2 mediated signal pathway.