Development of Bioactive Chemical Probes for Calcium-activated Chloride Channel

描述（由申请人提供）：离子通道是膜蛋白，存在于可兴奋细胞和非可兴奋细胞中。它们选择性地传导离子穿过细胞膜，并在细胞生理学中发挥关键作用，包括电和细胞信号传导、离子稳态和激素分泌。该提案的目的是找到编码钙激活氯离子通道（CaCC）的TMEM 16 A的有效和特异性小分子化学探针。CaCC的异常被认为是囊性纤维化、哮喘、慢性支气管炎和高血压等疾病的原因。其作用涉及许多生理过程，包括心脏和神经元兴奋、感觉转导、跨上皮分泌、平滑肌收缩和受精。 Lily Jan博士的实验室首次克隆了TMEM 16 A基因，我们与该实验室合作开发了几种细胞系和一种基于荧光的碘化物替代通量测定法。目前正在对CaCC靶标TMEM 16 A的高通量筛选方案进行测试。该提案概述了使用具有遗传编码的Cl-/I-生物传感器的TMEM 16 A表达细胞系进行> 300，000种化合物筛选以搜索CaCC的有效抑制剂/激活剂探针的计划。然后通过自动膜片钳记录评价活性化合物。与弗朗西斯科加州大学的Lily Jan博士合作，我们未来的计划包括分别在天然制剂中测试分离的化合物，如神经元和上皮细胞。这些活性化合物，在药理学和药物化学的进一步努力，可以被开发为工具或作为先导化合物，用于开发治疗药物的CaCC相关疾病。 公共卫生相关性：最近发现的钙激活氯离子通道（CaCC）为识别小分子探针铺平了新的道路。CaCC活性的异常被认为是导致几种主要疾病的原因，例如囊性纤维化、哮喘、慢性支气管炎和高血压。其作用涉及许多生理过程，包括心脏和神经元兴奋、感觉转导、跨上皮分泌、平滑肌收缩和受精。因此，特异性调节化合物可用于帮助治疗开发。

DESCRIPTION (provided by applicant): Ion channels are membrane proteins, found in both excitable cells and non-excitable cells. They selectively conduct ions across cellular membranes and play a critical role in cellular physiology, including electrical and cellular signaling, ion homeostasis, and hormone secretion. The objective of this proposal is to find potent and specific small molecule chemical probes for TMEM16A that encodes the calcium-activated chloride channel (CaCC). Abnormality of CaCC is thought to be causal to diseases, such as cystic fibrosis, asthma, chronic bronchitis, and hypertension. Its role is implicated in numerous physiological processes including cardiac and neuronal excitation, sensory transduction, trans-epithelial secretion, smooth muscle contraction, and fertilization. In collaboration with Dr. Lily Jan's laboratory, which first cloned the TMEM16A gene, we have developed several cell lines and a fluorescence-based iodide surrogate flux assay. The protocol for high-throughput screening is being tested for the CaCC target TMEM16A. This proposal outlines a plan to conduct a >300,000-compound screen using the TMEM16A expressing cell line with a genetically encoded Cl-/I- biosensor to search potent inhibitor/activator probes for CaCC. The active compounds will then be evaluated by automated patch-clamp recording. In collaboration with Dr. Lily Jan at University of California at San Francisco, our future plan includes testing of the isolated compounds in native preparations, such as neurons and epithelium, respectively. These active compounds, with further efforts in pharmacology and medicinal chemistry, may be exploited either as tools or as lead compounds for developing therapeutic remedies to the CaCC-associated diseases. PUBLIC HEALTH RELEVANCE: The recent discovery of the calcium-activated chloride channel (CaCC) has paved a new path to identify small molecule probes. Abnormality of CaCC activity is thought to be causal to several major diseases, such as cystic fibrosis, asthma, chronic bronchitis, and hypertension. Its role is implicated in numerous physiological processes including cardiac and neuronal excitation, sensory transduction, trans-epithelial secretion, smooth muscle contraction, and fertilization. Therefore, specific modulation compounds are useful to aid therapeutic development.