Human ion channel pharmacology in droplet bilayer membranes

液滴双层膜中的人体离子通道药理学

• 批准号: 8608543
• 负责人: Jason L. Poulos
• 金额: $ 54.45万
• 依托单位: LIBREDE, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8608543
• 关键词: Achievement; Address; Adoption; Agreement; Arrhythmia; Artificial Membranes; Automation; Biological Assay; Cardiac; Cardiac Death; Cell membrane; Cells; Contracts; Data; Data Quality; Development; Device or Instrument Development; Dose; Drug Interactions; Drug Targeting; Electrophysiology (science); Equipment; Guidelines; Housing; Human; Industry; Inhibitory Concentration 50; Ion Channel; Libraries; Literature; Market Research; Measurement; Measures; Membrane; Methods; Pain; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase; Physiological Processes; Preparation; Process; Process Measure; Protocols documentation; Reagent; Reporting; Research; Risk; Running; Safety; Simulate; Solutions; System; TRPV1 gene; Technology; Technology Transfer; Testing; Transmembrane Transport; Validation; Work; assay development; base; cost; drug discovery; high throughput screening; instrument; instrumentation; meetings; novel strategies; operation; patch clamp; prototype; public health relevance; relating to nervous system; scale up; screening; Achievement; Address; Adoption; Agreement; Arrhythmia; Artificial Membranes; Automation; Biological Assay; Cardiac; Cardiac Death; Cell membrane; Cells; Contracts; Data; Data Quality; Development; Device or Instrument Development; Dose; Drug Interactions; Drug Targeting; Electrophysiology (science); Equipment; Guidelines; Housing; Human; Industry; Inhibitory Concentration 50; Ion Channel; Libraries; Literature; Market Research; Measurement; Measures; Membrane; Methods; Pain; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase; Physiological Processes; Preparation; Process; Process Measure; Protocols documentation; Reagent; Reporting; Research; Risk; Running; Safety; Simulate; Solutions; System; TRPV1 gene; Technology; Technology Transfer; Testing; Transmembrane Transport; Validation; Work; assay development; base; cost; drug discovery; high throughput screening; instrument; instrumentation; meetings; novel strategies; operation; patch clamp; prototype; public health relevance; relating to nervous system; scale up; screening

DESCRIPTION (provided by applicant): Ion channel drug discovery and safety screening are limited by high cost per data point, low throughput, and complexity. To directly address these industry-wide pain points, Librede Inc. is developing an alternative cell-free technology for ion channel screening, based on the measurement of ion channels in artificial droplet membranes. By eliminating cells at the point of measurement, our technology enables much lower running costs and simplified assay development. In preliminary work, we have used our platform to measure TRPM8 and hERG ion channels and obtain drug IC50 and EC50 values in agreement with state-of-the-art ion channel electrophysiology platforms. In Phase I development of our technology, we created an automation- ready prototype array plate, and verified its reliability and high yield by using it to create over 1000 artificial membranes. In the Phase II work proposed here, we will validate our platform with four ion channels: hERG, Kv7.1, TRPV1, and Cav1.2, selected for their relevance to screening and their degree of complexity. We will optimize the methods of preparing these ion channels from commercially available host cells to achieve highly repeatable and reliable conductance measurements. We will then use these preparations to measure the effects of drugs on the channels' conductance in a dose-dependent manner, generating reliable and repeatable IC50 and EC50 values that we will compare to those obtained from automated patch clamp screening instrumentation. These drug measurements will be performed using the array plates we developed in Phase I, allowing us to evaluate their performance for a simulated screening application. During the proposed work, we will collaborate with the research group of Prof. Jacob Schmidt at UCLA who, with the PI, initially developed the proposed technology. The Schmidt group will aid Librede by initially processing and measuring the ion channel preparations and then transferring this technology to Librede to replicate, optimize, scale-up, and ultimately commercialize. At the completion of this work, we will have validated four highly relevant ion channels pharmacologically in our system and demonstrated its operation in assay conditions. This validation will demonstrate our system's performance to potential end users and allow placement of instrument prototypes with identified beta testers and early adopters. The Phase II work proposed here is a critical step in the development of our new cell-free electrophysiology platform which can address the main industry pain points of cost, throughput, and ease of use (Librede's value proposition), and has the potential to significantly impact ion channel drug discovery and safety screening.

描述（由申请人提供）：离子通道药物发现和安全筛查受每个数据点，低吞吐量和复杂性的高成本限制。为了直接解决这些范围内的行业疼痛点，Librede Inc.正在基于人造液滴膜中的离子通道的测量，为离子通道筛选开发了一种无细胞通道筛选的技术。通过在测量点消除细胞，我们的技术可以降低运行成本和简化的测定开发。在初步工作中，我们使用我们的平台来测量TRPM8和HERG离子通道，并获得与最先进的离子通道电生理学平台一致的药物IC50和EC50值。在技​​术的第一阶段开发中，我们创建了一个自动化的现有原型阵列板，并通过使用它来创建1000多个人造膜来验证其可靠性和高收益率。在此处提出的第二阶段工作中，我们将使用四个离子通道验证我们的平台：HERG，KV7.1，TRPV1和CAV1.2，以其与筛选及其复杂程度相关。我们将优化从市售宿主细胞中制备这些离子通道的方法，以实现高度可重复和可靠的电导测量。然后，我们将使用这些制剂以剂量依赖性的方式来测量药物对通道电导的影响，从而产生可靠且可重复的IC50和EC50值，我们将与从自动化贴片夹筛选仪器中获得的仪器进行比较。这些药物测量将使用我们在第一阶段开发的阵列板进行，使我们能够评估它们的性能以进行模拟筛选应用。在拟议的工作中，我们将与加州大学洛杉矶分校的雅各布·施密特（Jacob Schmidt）教授的研究小组合作，他最初与PI开发了该技术。 Schmidt组将通过最初处理和测量离子渠道准备工作，然后将该技术转移到Librede以复制，优化，扩展和最终商业化来帮助Librede。完成这项工作时，我们将在系统中在药理学上验证了四个高度相关的离子通道，并在测定条件下证明了其运行。该验证将证明我们的系统对潜在的最终用户的性能，并允许使用已确定的Beta测试仪和早期采用者放置仪器原型。此处提出的第二阶段工作是我们新的无细胞电生理平台开发的关键步骤，该平台可以解决主要行业的成本疼痛点，吞吐量和易用性（Librede的价值主张），并有可能显着影响离子通道药物发现和安全性筛查。