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蛋白酪氨酸磷酸酶SHP2(Src homology phosphotyrosyl phosphatase 2)参与了JAK/STAT、NF-κB和P13K/AKT等诸多信号途径的调控,与努南综合征、单核细胞白血病、骨髓增生异常综合征、B细胞急性淋巴细胞白血病和急性髓性白血病等疾病密切相关.因此,建立稳定可靠的体外SHP2抑制剂筛选模型,对于发现高效的化学小分子探针并深入研究相关机制,以及开发新药物治疗相关疾病具有重要意义.通过应用大肠杆菌系统克隆表达可溶的GST-SHP2融合蛋白,以矾酸钠(Na3VO4)为阳性抑制剂,本文建立基于96孔板的SHP2体外筛选模型,并对该模型的有效性进行评价.利用该筛选模型筛选了1 431个化合物,发现从植物山豆根(Sophora tonkinensis)中提取得到的化合物Shandougenine A对SHP2具有明显的抑制作用,进一步研究发现其浓度依赖性抑制SHP2活性,IC50值为(11.72±2.6)μmol/L.本文筛选模型的建立,为发现SHP2抑制剂提供了一条简便高效的途径,并为后续的机制研究及药物开发奠定了良好基础.图6表1参21
Src homology phosphotyrosyl phosphatase 2 is involved in the regulation of many signaling pathways such as JAK / STAT, NF-κB and P13K / AKT, and is associated with noonan syndrome, monocytic leukemia, myelodysplastic syndrome , B cell acute lymphoblastic leukemia and acute myeloid leukemia and other diseases.Therefore, the establishment of a stable and reliable in vitro SHP2 inhibitor screening model for the discovery of efficient chemical small molecule probes and in-depth study of the relevant mechanisms, as well as the development of new drug treatment Related diseases.Through the E. coli system cloning expression of soluble GST-SHP2 fusion protein, with sodium auric acid (Na3VO4) as a positive inhibitor, this paper established a 96-well-based SHP2 screening model in vitro, and the model 1 431 compounds were screened by this screening model and found that Shandougenine A extracted from plant Sophora tonkinensis had a significant inhibitory effect on SHP2 and further study found that its concentration-dependent inhibition SHP2 activity, the IC50 value was (11.72 ± 2.6) μmol / L. The establishment of the screening model in this paper provided the basis for the discovery of SHP2 inhibitors A simple and efficient way, and lay a good foundation for the follow-up mechanism research and drug development.