Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, AB, Canada.
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, AB, Canada; Unidad Radiofarmacia-Ciclotrón, División de Investigación, Facultad de Medicina, Universidad Nacional Autónoma de México, México City, Mexico.
Eur J Med Chem. 2021 Jan 1;209:112902. doi: 10.1016/j.ejmech.2020.112902. Epub 2020 Oct 10.
This research article describes an approach to modify the thiazolidinedione scaffold to produce test drugs capable of binding to, and inhibit, the in vitro transcriptional activity of the oncogenic protein FOXM1. This approach allowed us to obtain FOXM1 inhibitors that bind directly to the FOXM1-DNA binding domain without targeting the expression levels of Sp1, an upstream transcription factor protein known to activate the expression of FOXM1. Briefly, we modified the chemical structure of the thiazolidinedione scaffold present in anti-diabetic medications such as pioglitazone, rosiglitazone and the former anti-diabetic drug troglitazone, because these drugs have been reported to exert inhibition of FOXM1 but hit other targets as well. After the chemical synthesis of 11 derivatives possessing a modified thiazolidinedione moiety, we screened all test compounds using in vitro protocols to measure their ability to (a) dissociate a FOXM1-DNA complex (EMSA assay); (b) decrease the expression of FOXM1 in triple negative-breast cancer cells (WB assay); (c) downregulate the expression of FOXM1 downstream targets (luciferase reporter assays and qPCR); and inhibit the formation of colonies of MDA-MB-231 cancer cells (colony formation assay). We also identified a potential binding mode associated with these compounds in which compound TFI-10, one of the most active molecules, exerts binding interactions with Arg289, Trp308, and His287. Unlike the parent drug, troglitazone, compound TFI-10 does not target the in vitro expression of Sp1, suggesting that it is possible to design FOXM1 inhibitors with a better selectivity profile.
这篇研究文章描述了一种方法，可以修饰噻唑烷二酮骨架，生成能够与致癌蛋白 FOXM1 的体外转录活性结合并抑制其活性的测试药物。这种方法使我们能够获得 FOXM1 抑制剂，这些抑制剂直接与 FOXM1-DNA 结合域结合，而不针对 Sp1（一种已知激活 FOXM1 表达的上游转录因子蛋白）的表达水平。简而言之，我们修饰了存在于抗糖尿病药物中的噻唑烷二酮骨架的化学结构，如吡格列酮、罗格列酮和前抗糖尿病药物曲格列酮，因为这些药物已被报道具有抑制 FOXM1 的作用，但也会作用于其他靶点。在合成了 11 种具有修饰的噻唑烷二酮部分的衍生物后，我们使用体外方案筛选了所有测试化合物，以测量它们（a）解离 FOXM1-DNA 复合物的能力（EMSA 测定）；（b）降低三阴性乳腺癌细胞中 FOXM1 的表达（WB 测定）；（c）下调 FOXM1 下游靶标的表达（荧光素酶报告测定和 qPCR）；并抑制 MDA-MB-231 癌细胞集落的形成（集落形成测定）。我们还确定了与这些化合物相关的潜在结合模式，其中最活跃的分子之一 TFI-10 与 Arg289、Trp308 和 His287 发生结合相互作用。与母体药物曲格列酮不同，化合物 TFI-10 不针对 Sp1 的体外表达，这表明有可能设计出具有更好选择性特征的 FOXM1 抑制剂。
