Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation of Hubei Province, College of Pharmacy, Huazhong University of Science and Technology, HangKong Road, Wuhan, 430070, Hubei Province, China.
Cancer Chemother Pharmacol. 2012 Jun;69(6):1633-40. doi: 10.1007/s00280-012-1873-3. Epub 2012 May 8.
To assess the anti-angiogenesis potential and mechanism of RY10-4, a derivative of protoapigenone, which was verified the broad-spectrum anti-tumor activities by previous study.
RY10-4 and RY10-3 were synthesized according to the procedure described. Breast cancer cells MCF-7 and MDA-MB-231 that got the best performance in the previous anti-tumor activity screening were selected for further anti-cancer mechanism research. Firstly, cells proliferation assay of RY10-4 and RY10-3 was used to demonstrate the fact that the 4-hydroxy-2,5-cyclohexadien-1-one system would be the efficient pharmacophore of RY10-4. Then, a series of assays such as human umbilical vein endothelial cells (HUVECs) proliferation assay, HUVECs migration, tube network formation and morphological observations of zebrafish were applied to confirm its anti-angiogenesis activity. Upon RY10-4 treatment, the HIF-1α and VEGF were analyzed by western blot in normoxic and hypoxic conditions, meanwhile the PI3K-AKT-mTOR pathway-related protein such as AKT, p-AKT, mTOR and p-mTOR was also analyzed.
In the MCF-7, MDA-MB-231 and HUVECs proliferation assay, RY10-4 that has 4-hydroxy-2,5-cyclohexadien-1-one system showed distinct advantage compared with RY10-3. Tests had verified the anti-angiogenesis capability of RY10-4. Down-regulation of the HIF-1α and inhibition phosphorylation levels of AKT and mTOR were found to be the pathway that RY10-4 exerts its functions on anti-angiogenesis.
The structure of 4-hydroxy-2,5-cyclohexadien-1-one should be the effective pharmacophore of RY10-4. RY10-4 got fine performance in anti-tumor and anti-angiogenesis assay, and thus, the quinol compound will be the new hot-spot for further anti-tumor agency development.
评估原阿片碱衍生物 RY10-4 的抗血管生成潜力和机制，该化合物在之前的研究中已被证实具有广谱抗肿瘤活性。
根据先前描述的程序合成了 RY10-4 和 RY10-3。选择在前一次抗肿瘤活性筛选中表现最佳的乳腺癌细胞 MCF-7 和 MDA-MB-231 进行进一步的抗癌机制研究。首先，通过细胞增殖试验证明 RY10-4 的 4-羟基-2,5-环己二烯-1-酮系统将是 RY10-4 的有效药效团。然后，应用一系列试验，如人脐静脉内皮细胞（HUVECs）增殖试验、HUVECs 迁移试验、管网络形成和斑马鱼形态观察，来证实其抗血管生成活性。在 RY10-4 处理后，在常氧和低氧条件下通过 Western blot 分析 HIF-1α 和 VEGF，同时还分析了 PI3K-AKT-mTOR 通路相关蛋白，如 AKT、p-AKT、mTOR 和 p-mTOR。
在 MCF-7、MDA-MB-231 和 HUVECs 增殖试验中，具有 4-羟基-2,5-环己二烯-1-酮系统的 RY10-4 与 RY10-3 相比表现出明显的优势。试验验证了 RY10-4 的抗血管生成能力。发现下调 HIF-1α 和抑制 AKT 和 mTOR 的磷酸化水平是 RY10-4 发挥抗血管生成作用的途径。
4-羟基-2,5-环己二烯-1-酮的结构应是 RY10-4 的有效药效团。RY10-4 在抗肿瘤和抗血管生成试验中表现出色，因此，喹啉化合物将成为进一步开发抗肿瘤药物的新热点。
