Discovery of 4-aryl-4H-chromenes as a new series of apoptosis inducers using a cell- and caspase-based high-throughput screening assay.: 1.: Structure-activity relationships of the 4-aryl group
Discovery of 4-aryl-4H-chromenes as a new series of apoptosis inducers using a cell- and caspase-based high-throughput screening assay.: 1.: Structure-activity relationships of the 4-aryl group
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DOI：
10.1021/jm049640t
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发表时间：
2004-12-02
影响因子：
7.3
通讯作者：
Cai, SX
中科院分区：
文献类型：
--
作者：
Kemnitzer, W;Drewe, J;Cai, SX
通过应用新的基于细胞和半胱天冬酶的HTS测定，2-氨基-3-氰基-7-（二甲氨基）4-（3-甲氧基-4，5-亚甲二氧基苯基）-4H-色烯（1a）已被鉴定为有效的凋亡诱导剂。发现化合物Ia诱导核碎裂和PARP裂解，以及将细胞阻滞在G（2）/M期并诱导细胞凋亡，如通过流式细胞术分析测定在多种人细胞系（例如Jurkat、T47 D）中所确定的。通过对4-芳基的构效关系（SAR）研究，从筛选命中1a到先导化合物2-氨基-4-（2-氨基-2-氨基-4-（2-（3-溴-4，5-二甲氧基苯基）-3-氰基-7-（二甲基氨基）-4H-色烯（1c）和2-氨基-3-氰基-7-甲基-苯并吡喃（1c）。（二甲基氨基）4-（5-甲基-3-吡啶基）-4H-色烯（4 e），在T47 D乳腺癌细胞中的半胱天冬酶活化测定中的EC 50分别为19和11 nM。还发现2-氨基-4-芳基-3-氰基-7-（二甲基氨基）4 H-色烯在生长抑制MTT测定中具有高活性，化合物1c的GI（50）值在低纳摩尔范围内。值得注意的是，发现化合物Ie在紫杉醇抗性、P-糖蛋白过表达的MESSA/DX 5肿瘤细胞中具有2nM的GI（50）值。在功能上，发现化合物1c是微管蛋白聚合的有效抑制剂，并有效抑制秋水仙碱与微管蛋白的结合。这些结果证实了基于细胞的半胱天冬酶活化测定是发现有效的凋亡诱导剂的有力工具，并表明4-芳基-4H-色烯具有开发成未来抗癌剂的潜力。
By applying a novel cell- and caspase-based HTS assay, 2-amino-3-eyano-7-(dimethylamino)4-(3-methoxy-4,5-methylenedioxyphenyl)-4H-chromene (1a) has been identified as a potent apoptosis inducer. Compound la was found to induce nuclear fragmentation and PARP cleavage, as well as to arrest cells at the G(2)/M stage and to induce apoptosis as determined by the flow cytometry analysis assay in multiple human cell lines (e.g. Jurkat, T47D). Through structure-activity relationship (SAR) studies of the 4-aryl group, a 4- and 7-fold increase in potency was obtained from the screening hit la to the lead compounds 2-amino-4-(3-bromo-4,5-dimethoxyphenyl)-3-cyano-7-(dimethylamino)-4H-chromene (1c) and 2-amino-3-cyano-7-(dimethylamino)4-(5-methyl-3-pyridyl)-4H-chromene (4e), with an EC50 of 19 and 11 nM in the caspase activation assay in T47D breast cancer cells, respectively. The 2-amino-4-aryl-3-cyano-7-(dimethylamino)4H-chromenes also were found to be highly active in the growth inhibition MTT assay, with GI(50) values in the low nanomolar range for compound 1c. Significantly, compound le was found to have a GI(50) value of 2 nM in the paclitaxel resistant, p-glycoprotein overexpressed, MESSA/DX5 tumor cells. Functionally, compound 1c was found to be a potent inhibitor of tubulin polymerization and to effectively inhibit the binding of colchicine to tubulin. These results confirm that the cell-based caspase activation assay is a powerful tool for the discovery of potent apoptosis inducers and suggest that the 4-aryl-4H-chromenes have the potential to be developed into future anticancer agents.