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: 8251888
• 负责人: Jason L. Poulos
• 金额: $ 80.58万
• 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/8251888
• 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 Delivery Systems; Drug Formulations; Drug Interactions; 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; Screening procedure; 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

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. PUBLIC HEALTH RELEVANCE: Librede is developing a novel cell-free ion channel screening platform that has the potential to significantly reduce costs and increase throughput of drug discovery and safety screening for ion channels. We will develop an ion channel measurement workstation that is capable of completely automated processing and measurement of Librede's ion channel plates after the addition of drug compounds modulating the channel activity. We are validating this workstation by screening compounds for activity against the CLIC1 human ion channel which is thought to play an important role in Alzheimer's disease.

描述（由申请人提供）：离子通道在每个细胞中都是重要的药物靶标，它们在包括心脏周期和神经活动在内的广泛生理过程中起关键作用。结果，意外的药物与离子通道的相互作用也至关重要，需要对所有候选药物筛查特定的离子通道进行筛查。不幸的是，在高吞吐量中进行药物发现和安全筛选的测定离子通道是有问题的，因为必须将离子通道纳入细胞膜中，以允许测量其离子传输以确定其功能。结果，目前没有用于离子通道筛选的高质量，高通量测定。与传统药物筛查的可溶性蛋白质相比，自动贴片夹仪器的最新发展仍低于吞吐量的量级，还需要昂贵的仪器，专业的细胞系和消耗品。对于现有的离子通道筛选方法，信息质量，吞吐量和成本存在很大差距。 Librede的目标是开发新技术，以提高早期药物研究和开发的效率。为此，Librede正在开发一种使用人造细胞膜的离子通道筛选的替代性无细胞技术。 Librede的专利待定无单元的人造膜可以实现更高的吞吐量和更低的消耗成本，同时需要便宜的设备和训练有素的人员。 Librede由加州大学洛杉矶分校（UCLA）研究人员和该技术的发明者创立。在第一阶段，我们与多通道放大器同时测量了一个48个膜阵列板，并验证了测量性能等于或超过竞争性的自动化贴片夹仪器。在此处提出的第二阶段工作中，我们将通过将我们的I期仪器与流体处理和运动控制硬件集成在一起，以使用Librede的人造膜技术构建用于无细胞离子通道筛选的自动化工作站。我们将咨询我们工作站的设计，构建和操作中的药物筛查行业自动化专家，并确认它可以与我们以前的工作相似，可以处理和测量Librede的人造膜板。为了证明其能力，我们将执行有限的CLIC1屏幕，这是一个与阿尔茨海默氏病有关的离子通道。 CLIC1很难用常规的补丁夹筛选，这使得这是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组合作，他们以前已经证明了人造膜中Clic1的测量。我们将以完整的自动化模式运行工作站，测量离子通道活性是72个复合离子通道靶向药物库的药物浓度的函数。该演示本质上与行业屏幕基本相同，是Librede开发我们人工膜技术的主要里程碑，该技术有望改善离子通道筛选的成本和吞吐量的数量级。 公共卫生相关性：Librede正在开发一个新型的无细胞离子通道筛选平台，该平台有可能显着降低成本并增加药物发现和离子通道安全筛查的吞吐量。我们将开发一个离子通道测量工作站，该工作站能够在添加药物化合物调节通道活性后完全自动化的处理和测量。我们正在通过筛选对CLIC1人离子通道的活性的化合物来验证该工作站，该通道被认为在阿尔茨海默氏病中起着重要作用。