High throughput cell-free ion channel screening workstation

高通量无细胞离子通道筛选工作站

• 批准号: 8545181
• 负责人: Jason L. Poulos
• 金额: $ 53.93万
• 依托单位: LIBREDE, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-07 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8545181
• 关键词: Alzheimer' s Disease; Amplifiers; Arrhythmia; Artificial Membranes; Automation; Biological Assay; Businesses; California; Cardiac; Cardiac Death; Cell Culture Techniques; Cell Line; Cell membrane; Cells; Chloride Channels; Consult; Cultured Cells; Development; Drug Compounding; Drug Formulations; Drug Interactions; Drug Targeting; Equipment; Freezing; Goals; Human; Human Resources; Industry; Ion Channel; Lead; Legal patent; Libraries; Licensing; Liquid substance; Los Angeles; Measurement; Measures; Membrane; Methods; Molecular; Motion; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological Processes; Play; Preclinical Drug Evaluation; Process; Process Measure; Proteins; Research; Research Personnel; Resource Sharing; Risk; Role; Running; Safety; Shipping; Ships; Solutions; Staging; System; Technology; Training; Transmembrane Transport; Universities; Water; Work; assay development; base; cell type; cost; design; design and construction; drug candidate; drug discovery; high throughput screening; improved; instrument; instrumentation; new technology; novel; operation; patch clamp; professor; prototype; relating to nervous system; research and development; screening

DESCRIPTION (provided by applicant): Ion channels are important drug targets-present in every cell, they play key roles in a wide range of physiological processes including the cardiac cycle and neural activity. As a result, unintended drug interactions with ion channels are also of critical importance, requiring the screening of all drug candidates against specific ion channels. Unfortunately, assaying ion channels for pharmaceutical discovery and safety screening is problematic to perform in high throughput because the ion channels must be incorporated into a cell membrane to allow measurement of their ionic transport to determine their functionality. As a result, there are currently no high quality, high throughput assays for ion channel screening. Recent developments of automated patch clamp instrumentation are still over an order of magnitude lower throughput than conventional drug screening for soluble proteins and also require expensive instrumentation, specialized cell lines, and consumables. For existing methods of ion channel screening, there is a large gap in information quality, throughput, and cost. Librede's goal is to develop new technologies that increase the efficiency of early stage pharmaceutical research and development. To this end, Librede is developing an alternative cell-free technology for ion channel screening using artificial cell membranes. Librede's patent pending formulation of cell-free artificial membranes can enable higher throughput and lower consumable costs while requiring less expensive equipment and trained personnel. Librede was founded by UCLA researchers and the inventors of this technology. In Phase I, we measured a 48 membrane array plate simultaneously with a multichannel amplifier and verified that the measurement performance was equal to or exceeded competitive automated patch clamp instruments. In the Phase II work proposed here, we will build on this work by integrating our Phase I instrumentation with fluid handling and motion control hardware to construct an automated workstation for cell-free ion channel screening using Librede's artificial membrane technology. We will consult with a pharmaceutical screening industry automation expert in the design, construction, and operation of our workstation and confirm that it can process and measure Librede's artificial membrane plates similarly to our previous work. To demonstrate its capabilities, we will perform a limited screen of CLIC1, an ion channel implicated in Alzheimer's disease. CLIC1 is particularly difficult to screen with conventional patch clamp, which makes this an ideal proof-of-concept application of our platform. We will work with the Schmidt group at UCLA, who have previously demonstrated the measurement of CLIC1 in artificial membranes. We will run the workstation in full automation mode, measuring ion channel activity as a function of drug concentration for a 72 compound ion channel-targeted drug library. This demonstration is essentially identical to an industry screen and is a major milestone for Librede's development of our artificial membrane technology, which promises an order of magnitude improvement in cost and throughput for ion channel screening.

描述（由申请人提供）：离子通道是重要的药物靶标-存在于每个细胞中，它们在包括心动周期和神经活动在内的广泛生理过程中发挥关键作用。因此，与离子通道的非预期药物相互作用也至关重要，需要针对特定离子通道筛选所有候选药物。不幸的是，测定用于药物发现和安全筛选的离子通道在高通量下进行是有问题的，因为离子通道必须被掺入细胞膜中以允许测量它们的离子转运以确定它们的功能。因此，目前没有用于离子通道筛选的高质量、高通量测定。自动化膜片钳仪器的最新发展仍然比用于可溶性蛋白的常规药物筛选的通量低一个数量级，并且还需要昂贵的仪器、专门的细胞系和消耗品。对于现有的离子通道筛选方法，在信息质量、通量和成本方面存在很大差距。Librede的目标是开发新技术，提高早期药物研发的效率。为此，Librede正在开发一种替代的无细胞技术，用于使用人工细胞膜进行离子通道筛选。Librede正在申请专利的无细胞人工膜配方可以实现更高的吞吐量和更低的消耗成本，同时需要更便宜的设备和训练有素的人员。Librede是由加州大学洛杉矶分校的研究人员和这项技术的发明者创立的。在第一阶段，我们用多通道放大器同时测量了48个膜阵列板，并验证了测量性能等于或超过竞争性自动膜片钳仪器。在这里提出的第二阶段工作中，我们将通过将我们的第一阶段仪器与流体处理和运动控制硬件集成来构建一个自动化工作站，用于使用Librede的人工膜技术进行无细胞离子通道筛选。我们将与制药筛选行业的自动化专家就我们工作站的设计、建造和操作进行咨询，并确认它可以像我们以前的工作一样处理和测量Librede的人工膜板。为了证明其能力，我们将进行CLIC 1的有限筛选，CLIC 1是一种与阿尔茨海默病有关的离子通道。CLIC 1特别难以用传统的膜片钳进行筛选，这使得它成为我们平台的理想概念验证应用。我们将与加州大学洛杉矶分校的施密特小组合作，他们以前曾演示过在人工膜中测量CLIC 1。我们将在全自动模式下运行工作站，测量离子通道活性作为72化合物离子通道靶向药物库的药物浓度的函数。该演示与工业屏幕基本相同，是Librede开发人工膜技术的一个重要里程碑，该技术有望在离子通道筛选的成本和通量方面实现数量级的改进。