DataSheet1_Angiogenesis, Anti-Tumor, and Anti-Metastatic Activity of Novel α-Substituted Hetero-Aromatic Chalcone Hybrids as Inhibitors of Microtubule Polymerization.pdf

A library of new heteroaromatic ring-linked chalcone analogs were designed and synthesized of these, compound 7m with α-CH3 substitution and bearing a benzofuran ring, displaying the most potent activity, with IC50 values of 0.07–0.183 µM against three cancer cells. Its low cytotoxicity toward normal human cells and strong potency on drug-resistant cells revealed the possibility for cancer therapy. It also could moderately inhibit in vitro tubulin polymerization with an IC50 value of 12.23 µM, and the disruption of cellular architecture in MCF-7 cells was observed by an immunofluorescence assay. Cellular-based mechanism studies elucidated that 7m arrested the cell cycle at the G2/M phase and induced apoptosis by regulating the expression levels of caspases and PARP protein. Importantly, the compound 7 m was found to inhibit HUVEC tube formation, migration, and invasion in vitro. In vivo assay showed that 7m could effectively destroy angiogenesis of zebrafish embryos. Furthermore, our data suggested that treatment with 7m significantly reduced MCF-7 cell metastasis and proliferation in vitro and in zebrafish xenograft. Collectively, this work showed that chalcone hybrid 7m deserves further investigation as dual potential tubulin polymerization and angiogenesis inhibitor.

本研究设计并合成了一系列新型杂芳环（heteroaromatic ring）连接的查尔酮类似物（chalcone analogs）库。其中，带有α-甲基取代（α-CH3 substitution）且含苯并呋喃环（benzofuran ring）的化合物7m展现出最强的抗肿瘤活性，对三种癌细胞的半抑制浓度（IC50）为0.07–0.183 µM。该化合物对正常人体细胞毒性较低，且对耐药癌细胞具有优异的抑制活性，提示其具备用于癌症治疗的潜力。此外，该化合物可适度抑制体外微管蛋白聚合（tubulin polymerization），其半抑制浓度为12.23 µM；通过免疫荧光测定（immunofluorescence assay）可观察到，该化合物会破坏MCF-7细胞的细胞骨架结构。细胞水平的机制研究阐明，化合物7m可将细胞周期阻滞于G2/M期，并通过调控半胱天冬酶（caspases）与聚ADP核糖聚合酶（PARP）蛋白的表达水平诱导细胞凋亡（apoptosis）。尤为重要的是，研究发现化合物7m可在体外抑制人脐静脉内皮细胞（Human Umbilical Vein Endothelial Cells, HUVEC）的管腔形成、迁移与侵袭。体内实验结果显示，化合物7m可有效破坏斑马鱼胚胎（zebrafish embryos）的血管生成（angiogenesis）过程。此外，实验数据表明，经化合物7m处理后，MCF-7细胞的增殖与转移能力在体外及斑马鱼异种移植模型中均受到显著抑制。综上，本研究证实，查尔酮杂合物7m兼具微管蛋白聚合抑制与血管生成抑制双重活性，具备进一步开发研究的价值。