Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 84084, Fisciano, Salerno, Italy.
Department of Cell Physiology and Metabolism, University of Geneva, Rue Michel-Servet 1, CH - 1211, Geneva, Switzerland.
Eur J Med Chem. 2021 Dec 15;226:113872. doi: 10.1016/j.ejmech.2021.113872. Epub 2021 Sep 25.
Based on biological results of previous synthesized pyrazolyl ureas able to interfere with angiogenesis process, we planned and synthesized the new benzyl-urea derivatives 2-4; some of them showed an interesting anti-proliferative profile and particularly 4e potently inhibited HUVEC proliferation. To shed light on the mechanism of action of 4e, its interactome has been deeply inspected to identify the most prominent protein partners, mainly taking into account kinome and phosphatome, through drug affinity responsive target stability experiments, followed by targeted limited proteolysis analysis. From these studies, PP1γ emerged as the most reliable 4e potential target in HUVEC. Molecular docking simulations on PP1γ were carried out to predict 4e binding mode. To assess its potential anti-angiogenic effect, 4e was tested in vitro to verify interference on kinase and phosphate activities. Overall, our results evidenced for 4e an interesting anti-angiogenic action, probably due to its action at intracellular level on PP1γ signalling pathways.
基于先前能够干扰血管生成过程的合成吡唑基脲的生物学结果，我们设计并合成了新的苄基脲衍生物 2-4；其中一些表现出有趣的抗增殖特性，特别是 4e 能够强烈抑制 HUVEC 的增殖。为了阐明 4e 的作用机制，我们深入研究了其相互作用组，以鉴定最突出的蛋白质伙伴，主要考虑到激酶组和磷酸酶组，通过药物亲和反应靶标稳定性实验，随后进行靶向有限蛋白酶分析。从这些研究中，PP1γ 作为 HUVEC 中最可靠的 4e 潜在靶标脱颖而出。对 PP1γ 进行了分子对接模拟，以预测 4e 的结合模式。为了评估其潜在的抗血管生成作用，在体外测试了 4e 以验证对激酶和磷酸酶活性的干扰。总的来说，我们的结果表明 4e 具有有趣的抗血管生成作用，可能是由于其在细胞内水平对 PP1γ 信号通路的作用。
