Abstract：
Novel BODIPY-estradiol conjugates have been synthesized by selecting position C-3-O for labeling. The conjugation strategy was based on Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) or etherification. Estradiol derivatives used as azide partners bearing an ω-azidoalkyl function through C4-C8-long linkers have been prepared. CuAAC reactions of estradiol azides with BODIPY alkyne furnished fluorescent 3-O-labeled conjugates bearing the triazole ring as a coupling moiety. Williamson etherifications of 3-O-(ω-bromoalkyl)-17β-estradiol derivatives with BODIPY-OH resulted in labeled conjugates connected with an ether moiety. Interactions of the conjugates with estrogen receptor (ER) were investigated using molecular docking calculations in comparison with estradiol. The conjugates occupied both the classical and alternative binding sites on human ERα, with slightly lower binding affinity to references estradiol and diethystilbestrol. All compounds have displayed reasonable estrogenic activity. They increased the proliferation of ER-positive breast cancer cell line MCF7 contrary to ER-negative SKBR-3 cell line. The most potent compound 13a induced the transcriptional activity of ER in dose-dependent manner in dual luciferase recombinant reporter model and increased progesterone receptor\'s expression, proving the retained estrogenic activity. The fluorescence of candidate compound 13a co-localised with the ERα. The newly synthesized labeled compounds might serve as good starting point for further development of fluorescent probes for modern biological applications. In addition to studying steroid uptake and transport in cells, e.g. in the processes of biodegradation of estrogen-hormones micropollutants, they could also be utilized in examination of estrogen-binding proteins.
摘要：
已经通过选择用于标记的位置C-3-O合成了新的BODIPY-雌二醇缀合物。缀合策略基于Cu(I)催化的叠氮化物-炔环加成(CuAAC)或醚化。已经制备了通过C4-C8长接头用作带有ω-叠氮烷基官能团的叠氮化物伴侣的雌二醇衍生物。雌二醇叠氮化物与BODIPY炔烃的CuAAC反应提供了带有三唑环作为偶联部分的荧光3-O标记的缀合物。3-O-（ω-溴烷基）-17β-雌二醇衍生物与BODIPY-OH的Williamson醚化产生与醚部分连接的标记缀合物。与雌二醇相比，使用分子对接计算研究了缀合物与雌激素受体（ER）的相互作用。缀合物占据了人ERα上的经典和替代结合位点，对参考雌二醇和己烯雌酚的结合亲和力略低。所有化合物都显示出合理的雌激素活性。与ER阴性SKBR3细胞系相反，它们增加了ER阳性乳腺癌细胞系MCF7的增殖。最有效的化合物13a在双荧光素酶重组报告子模型中以剂量依赖性方式诱导ER的转录活性，并增加孕激素受体的表达，证明保留的雌激素活性。候选化合物13a的荧光与ERα共定位。新合成的标记化合物可能是进一步开发用于现代生物学应用的荧光探针的良好起点。除了研究类固醇在细胞中的摄取和转运，例如，在雌激素-激素微污染物的生物降解过程中，它们也可用于检测雌激素结合蛋白。
