Phenotypic screening is a neoclassical approach for drug discovery. We conducted phenotypic screening for insulin secretion enhancing agents using INS-1E insulinoma cells as a model system for pancreatic beta-cells. A principal regulator of insulin secretion in beta-cells is the metabolically regulated potassium channel Kir6.2/SUR1 complex. To characterize hit compounds, we developed an assay to quantify endogenous potassium channel activity in INS-1E cells. We quantified ligand-regulated potassium channel activity in INS-1E cells using fluorescence imaging and thallium flux. Potassium channel activity was metabolically regulated and coupled to insulin secretion. The pharmacology of channel opening agents (diazoxide) and closing agents (sulfonylureas) was used to validate the applicability of the assay. A precise high-throughput assay was enabled, and phenotypic screening hits were triaged to enable a higher likelihood of discovering chemical matter with novel and useful mechanisms of action.

中文翻译：

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胰岛 β 细胞钾通道的高通量检测：用于从表型筛选中鉴定的胰岛素促分泌剂的药理学表征

表型筛选是一种新古典药物发现方法。我们使用 INS-1E 胰岛素瘤细胞作为胰腺 β 细胞的模型系统对胰岛素分泌增强剂进行了表型筛选。β 细胞中胰岛素分泌的主要调节器是代谢调节的钾通道 Kir6.2/SUR1 复合物。为了表征命中化合物，我们开发了一种测定方法来量化 INS-1E 细胞中的内源性钾通道活性。我们使用荧光成像和铊通量量化了 INS-1E 细胞中配体调节的钾通道活性。钾通道活性受代谢调节并与胰岛素分泌相关。通道开放剂（二氮嗪）和封闭剂（磺酰脲类）的药理学用于验证该测定的适用性。启用了精确的高通量检测，