Department of Inorganic Chemistry, Faculty of Chemistry, Gdansk University of Technology, Gdansk, Poland.
Department of Organic Chemistry, Faculty of Chemistry, Gdansk University of Technology, Gdansk, Poland.
Drug Dev Res. 2019 Sep;80(6):857-866. doi: 10.1002/ddr.21569. Epub 2019 Jul 13.
In the present work, we described convenient methods for the synthesis of N-thiophosphorylated 3-(4-aminophenyl)-coumarin-7-O-sulfamates as steroid sulfatase (STS) inhibitors. To design the structures of the potential STS inhibitors, molecular modeling techniques were used. A computational docking method was used to determine the binding modes of the synthesized inhibitors as well as to identify potential interactions between specified functional groups on the inhibitors and the amino acid residues present in the active site of the enzyme. The inhibitory activities of the synthesized compounds were tested in an enzymatic assay with STS isolated from a human placenta. Within the set of newly synthesized compounds, 9e demonstrated the highest inhibitory activity in the enzymatic assay with an IC value of 0.201 μM (the IC value of 667-COUMATE in the same test was 0.062 μM). Furthermore, we tried to verify if the obtained STS inhibitors are able to pass through the cellular membrane effectively in cell line experiments. In the course of our study, we determined the STS activity in the MCF-7 cell line after incubation in the presence of the inhibitors (at 100 nM concentration). For this evaluation, we included newly synthesized compounds 9a-g and their N-phosphorylated analogs 6a-h, whose synthesis has been previously described. We found that the lowest STS activities were measured in the presence of N-phosphorylated derivatives 6e (0.1% of STS activity) and 6f (0.2% of STS activity). The measured STS activity in the presence of 667-COUMATE (used as a reference) was 0.1%. Moreover, at concentrations up to 1 μM, the most active compounds (6e, 6f, 9b, and 9e) did not exert any toxic effects on zebrafish embryos.
在本工作中，我们描述了方便的方法，用于合成 N-硫代膦酰基 3-(4-氨基苯基)-香豆素-7-O-磺胺酯作为甾体硫酸酯酶 (STS) 抑制剂。为了设计潜在 STS 抑制剂的结构，使用了分子建模技术。计算对接方法用于确定合成抑制剂的结合模式，并确定抑制剂上指定官能团与酶活性位点中存在的氨基酸残基之间的潜在相互作用。使用从人胎盘分离的 STS 在酶测定中测试了合成化合物的抑制活性。在所合成的化合物中，9e 在酶测定中表现出最高的抑制活性，IC 值为 0.201 μM（在相同测试中，667-COUMATE 的 IC 值为 0.062 μM）。此外，我们试图在细胞系实验中验证所获得的 STS 抑制剂是否能够有效地穿过细胞膜。在我们的研究过程中，我们在 MCF-7 细胞系中测定了在抑制剂存在下（在 100 nM 浓度下）的 STS 活性。为了进行此评估，我们包括了新合成的化合物 9a-g 及其 N-磷酸化类似物 6a-h，其合成先前已被描述。我们发现，在 N-磷酸化衍生物 6e（STS 活性的 0.1%）和 6f（STS 活性的 0.2%）存在下测量的 STS 活性最低。在 667-COUMATE（用作参考）存在下测量的 STS 活性为 0.1%。此外，在高达 1 μM 的浓度下，最活跃的化合物（6e、6f、9b 和 9e）对斑马鱼胚胎没有任何毒性作用。
