Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009, China.
Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China.
Bioorg Chem. 2020 May;98:103150. doi: 10.1016/j.bioorg.2019.103150. Epub 2019 Aug 1.
β-Sitosterols, is a common steroid that can be identified in a variety of plants and their efficacy in promoting wound healing has been demonstrated. Na/K-ATPase, more than a pump, its signal transduction function for involvement in cell growth regulation attracts widespread concern. The Na/K-ATPase/Src receptor complex can serve as a receptor involved in multiple signaling pathways including promoting wound healing pathways. To finding potent accelerating wound healing small molecular, we choose the high inhibitory activity of Na/K-ATPase and non-cardiotoxic natural compound, β-sitosterol as the substrate. A series of β-sitosterol derivatives were designed, synthesized and evaluated as potential Na/K-ATPase inhibitors. Among them, compounds 31, 47, 49, showed improved inhibitory activity on Na/K-ATPase, with IC value of 3.0 μM, 3.4 μM, 2.2 μM, which are more potent than β-sitosterol with IC 7.6 μM. Especially, compound 49 can induce cell proliferation, migration and soluble collagen production in L929 fibroblasts. Compared to model, compound 49 can accelerate wound healing in SD rats. Further studies indicated that 49 can activate the sarcoma (Src), uptake the protein kinase B (Akt), extracellular signal-regulated kinase (ERK) proteins expression in a concentration dependent manner. Finally, binding mode of compound 49 with Na/K-ATPase was studied, which provides insights into the determinants of potency and selectivity. These results proved β-stitosterol derivative 49 can serve as an effective inhibitor of Na/K-ATPase and potential candidate for accelerating wound healing agents.
β-谷甾醇是一种常见的甾体，可以在多种植物中被识别，其促进伤口愈合的功效已得到证实。Na/K-ATP 酶，不仅仅是一种泵，其参与细胞生长调节的信号转导功能引起了广泛关注。Na/K-ATP 酶/Src 受体复合物可以作为一种参与多种信号通路的受体，包括促进伤口愈合通路。为了寻找有效的促进伤口愈合的小分子，我们选择了对 Na/K-ATP 酶具有高抑制活性且无心脏毒性的天然化合物β-谷甾醇作为底物。设计、合成并评价了一系列β-谷甾醇衍生物，作为潜在的 Na/K-ATP 酶抑制剂。其中，化合物 31、47、49 对 Na/K-ATP 酶的抑制活性提高，IC 值分别为 3.0μM、3.4μM、2.2μM，比β-谷甾醇（IC 7.6μM）更有效。特别是化合物 49 可以诱导 L929 成纤维细胞的增殖、迁移和可溶性胶原产生。与模型相比，化合物 49 可以加速 SD 大鼠的伤口愈合。进一步的研究表明，49 可以激活肉瘤（Src），并呈浓度依赖性摄取蛋白激酶 B（Akt）、细胞外信号调节激酶（ERK）蛋白表达。最后，研究了化合物 49 与 Na/K-ATP 酶的结合模式，为了解效力和选择性的决定因素提供了线索。这些结果证明β-谷甾醇衍生物 49 可以作为 Na/K-ATP 酶的有效抑制剂和促进伤口愈合的潜在候选药物。
