靶向ERK新型抗乳腺癌小分子BL-EI001诱导自噬性死亡的机制研究

Mitogen-activated protein kinase (MAPK) signaling plays its key role in breast cancer development, which has been regarded as one of the most potential cancer targets in breast cancer therapy. In our previous studies, we designed and synthezied a totally new small-molecule compound named BL-EI001 with anti-proliferative activities targeting ERK. We found that BL-EI001 could induce death-receptor and mitochondrial apoptosis in breast cancer cells. In addition, we showed that BL-EI001 bear a good anti-tumor efficacy and had a therapeutic potent on mouse model in vivo. However, the how BL-EI001 may target ERK still remains to be clarified. As mentioned above, the aim of this study focuses on elucidating how BL-EI001 specifically binds to its protein target ERK, regulates target gene experssion and inhibits p-ERK at translational level. Our current study will explore how BL-EI001 regulates ERK-related signaling pathways, induces autophagic cell death via ERK-eEF2K axis, and thus eventually triggering apoptosis in vitro and in vivo. In summary, we will conclude the mechanisms of BL-EI001-induced autophagic cell death and its specially targeted function of ERK. These findings will provide a new clue on exploring BL-EI001 as a novel anti-tumor drug for the future breasr cancer therapuetics.

丝裂原活化蛋白激酶（MAPK）信号在乳腺癌发生发展中起重要作用，已成最具潜力抗乳腺癌药研发靶标之一。我们前期工作以MAPK级联通路中的ERK为靶标，通过药物设计合成及活性筛选得到结构全新的小分子BL-EI001，发现它诱导乳腺癌细胞发生死亡受体和线粒体途径的凋亡，且在小鼠模型中具有较好的抗肿瘤活性和潜在治疗作用。然而BL-EI001的靶向作用机制仍然不是十分清楚。基于前期的工作基础，本项目旨在阐明BL-EI001与其靶点ERK作用靶向性和专一性，小分子与靶蛋白的结合功能；小分子影响靶基因表达，抑制靶蛋白磷酸化等。探讨BL-EI001如何通过与ERK有关的分子信号，包括调控自噬性死亡的新通路“ERK-eEF2K”而引发的凋亡，以及它在体内外乳腺癌模型中发挥的重要作用。本研究将有助于确认BL-EI001是否作为抗肿瘤特异性靶向化合物及其作用基础,为开发新型靶向乳腺癌小分子抑制剂提供新思路。

丝裂原活化蛋白激酶（MAPK）信号在乳腺癌发生发展中起重要作用，已成最具潜力抗乳腺癌药研发靶标之一。我们前期工作以MAPK级联通路中的ERK为靶标，通过药物设计合成及活性筛选得到结构全新的小分子BL-EI001，发现它诱导乳腺癌细胞发生死亡受体和线粒体途径的凋亡，且在小鼠模型中具有较好的抗肿瘤活性和潜在治疗作用。基于前期的工作基础，我们通过形态学观察、免疫荧光法、流式细胞术、免疫印迹法等手段对BL-EI001在乳腺癌细胞中诱导自噬的分子机制进行了研究，通过iTRAQ蛋白组学技术分析了BL-EI001通过ERK-eEF2K通路调控的分子机制。通过分子对接模式分析和分子动力学模拟对BL-EI001及其优化后的衍生物22ac的靶向性进行了分析，并采用激酶组学技术对优化后的衍生物22ac的专一性进行了研究，22ac对ERK1/2具有较好的把向专一性，并采用异种植瘤裸鼠模型对BL-EI001及其优化后的衍生物进行了抗肿瘤作用及机制研究。本研究可以为开发新型靶向乳腺癌小分子抑制剂提供新思路。