蛋白酪氨酸磷酸酶SHP2抑制剂体外筛选模型的建立及应用  

Establishment and application of in vitro screening platform for inhibitors of protein tyrosine phosphate SHP2

作　　者：邓腾[1,2] 杨莉娟[1] 罗国勇 李晟[1,2] 杜宝文[1] 王飞[1] 

机构地区：[1]中国科学院成都生物研究所,成都610041 [2]中国科学院大学,北京100049

出　　处：《应用与环境生物学报》2015年第4期602-606,共5页Chinese Journal of Applied and Environmental Biology

基　　金：四川省科技厅基础支撑计划项目(2012SZ0219);四川省青年科技基金项目(2014JQ0028)资助~~

摘　　要：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.The protein tyrosine phosphatase SHP2 （Src homology phosphotyrosyl phosphatase 2, encoded by PTPN11 gene） is widely expressed and involved in multiple cell signaling processes, such as the JAK/STAT, NF-κB and P13K/AKT pathways. Mutations of PTPN11 have been found in Noonan syndrome, juvenile myelomonocytic leukemia （JMML）, and several types of human malignancies. In order to discover potent small molecules regulating SHP2, we set up an in vitro screening platform for SHP2 based on GST-SHP2 fusion protein expressed by E. coil expression system. This model was validated using Na3VO4 as the positive inhibitor; a compound library with 1 431 natural or synthetic compounds was screened. This effort led to the identification of Shandougenine A, isolated from Sophora tonkinensis, displaying potent inhibitory activity toward SHP2 protein with the IC50 value of 11.72±2.6 μmol/L. The establishment of this in vitro screening model provides an effective and convenient platform to find novel SHP2 inhibitors, which may be used as potential probes for investigating SHP2-ralated mechanisms or drug candidates for the treatment of SHP2-related diseases.

关 键 词：SHP2 抑制剂 体外筛选 Shandougenine A 

分 类 号：R363[医药卫生—病理学]