由内质网钙感受器STIM和质膜钙离子通道Orai1所共同介导的钙池操纵钙内流（Store-Operated Calcium Entry，SOCE）是一种普遍存在的细胞钙信号和钙稳态调控途径。SOCE与肿瘤、心血管、神经及免疫系统多种疾病密切相关。而大多数现存的SOCE工具药存在着起效慢，特异性或可逆性差等缺点，因而为探究SOCE激活的分子机制并为SOCE相关疾病的治疗提供新的思路，研发效果可逆的高度特异的SOCE工具药势在必行。 本研究首先建立了一个灵敏高效的药物筛选系统。该系统包括以SOCE相关蛋白—Orai1为靶蛋白的高效的虚拟药物筛选平台，一个用于检测SOCE的稳定表达高灵敏度的钙指示蛋白GCaMP6m的细胞平台，以及一个检测药物对GCaMP6m指示的SOCE的影响的高内涵药物筛选平台。本研究首先通过针对Orai1的分子对接计算，从ZINC数据库中11,340,974个“类药”小分子找到了129种与可能Orai1几个关键活性区域紧密结合的候选小分子药物。之后在高内涵药物筛选平台上用GCaMP6m细胞对这些类药小分子，以及不同来源的73种中草药活性成分单体进行了初筛，从中获得对GCaMP6m细胞中SOCE信号抑制作用较强的9种小分子化合物和4种中草药物活性成分单体。最后通过单细胞水平的荧光钙及FRET成像技术，对上述初筛得到的药物进行了进一步的筛选，从中分别得到了一个抑制SOCE效果最好的小分子化合物YJW144和中草药活性物质单体ZQS11。 结合分子生物学、激光共聚焦等技术，对上述两个药物及实验室前期获得的一个强心苷类药物digitoxin的作用机制进行了进一步的鉴定。结果表明这三种药物均是直接作用在Orai1离子通道上，并且它们的抑制作用与Orai1通道口的结构有关。这些结果暗示Orai1是一个非常好的SOCE工具药研发靶标。 本研究筛选并初步鉴定的几个SOCE工具药物将为探究SOCE激活的分子机制及治疗SOCE相关疾病提供新的思路。而上述结果也暗示，本研究所建立的高效，高灵敏度的药物筛选平台还有望进行各种钙信号调控药物及调控基因的高效筛选，具有非常好的应用前景。

Store-Operated Calcium（Ca2+）Entry（SOCE）is mediated by the stromal interaction molecule （STIM） and the Orai1 Ca2+ channel. SOCE is a ubiquitous Ca2+ signalling pathway, crucial for Ca2+ homeostasis. SOCE is closely related to many types of diseases such as tumor, and diseases in cardiovascular, nervous and immune system. However, most available SOCE inhibitors have flaws and fail in one or more of the following criteria: avidity, specificity, rapidity of action, and reversibility. To further dissect the underlying molecular mechanism of SOCE and provide new ideas for the treatment of SOCE related diseases, it is crucial to discover and design new pharmacological tools. We thus established a sensitive and efficient drug screening system. The system includes an efficient virtual drug screening platform using Orai1 as a target, a stable cell platform stably expressing GCaMP6m, a highly sensitive Ca2+indciator for measuring SOCE, and a high-content imaging system. The results showed that 9 of the small molecular compounds and 4 Chinese herbal monomers could inhibit SOCE with a tens of micro Mole concentration. After further tests on the above-mentioned 13 drugs with single-cell Ca2+ and FRET imaging, YJW144, a small molecule compound, and ZQS11, a Chinese herbal monomer, showed best inhibitory effects on SOCE. The molecular mechanism underlying SOCE inhibition of the above two drugs and digitoxin, a less characterized SOCE inhibitor found in our previous research, were further examined. The results showed that all three drugs could directly acting on the Orai1 channel, and their inhibitory effects were related to the structure of the Orai1 pore. These results indicate thatOrai1 is a very good target for the development of SOCE inhibitors. Overall, we screened and identified several SOCE inhibitors, which may help dissecting the molecular mechanism underlying SOCE activation and the treatments of SOCE related diseases. Also, the highly efficient and sensitive drug-screening platform established in this study is expected to find more applications for the screening of more compounds or genes that could regulate Ca2+ signalling and homeostasis.