Development of a chemical reaction sensor array platform for label-free, real-time kinetic analysis of enzyme-substrate reactions to enable high-throughput drug discovery

开发化学反应传感器阵列平台，用于酶-底物反应的无标记实时动力学分析，以实现高通量药物发现

• 批准号: 10677748
• 负责人: Bharath Takulapalli
• 金额: $ 102.47万
• 依托单位: SPOC PROTEOMICS INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-05 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10677748
• 关键词: Acoustics; Alzheimer' s Disease; Alzheimer' s disease pathology; Area; Automation; Binding; Biochemical Reaction; Biological Assay; Biosensor; Charge; Coupled; Custom; Data; Detection; Development; Disease; Drug Screening; Drug Targeting; Drug usage; Electronics; End Point Assay; Enzyme Activators; Enzyme Inhibition; Enzyme Inhibitor Drugs; Enzymes; Expenditure; Fluorescence; Future; Goals; Homeostasis; In Vitro; Ions; Kinetics; Label; Libraries; Link; Liquid substance; Literature; MAPT gene; Measurement; Measures; Mediating; Metabolism; Methods; Monitor; Noise; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Play; Protein Dephosphorylation; Publishing; Reaction; Reporting; Reproducibility; Role; Scanning; Semiconductors; Signal Transduction; Surface; System; Technology; Testing; Time; Transistors; Validation; Variant; assay development; candidate selection; chemical reaction; cost; data acquisition; detection assay; detection platform; detection sensitivity; drug discovery; drug testing; electronic sensor; enzyme activity; enzyme substrate; fabrication; glycogen synthase kinase 3 beta; high throughput screening; high-throughput drug screening; inhibitor; luminescence; multiplex detection; nano; nanosensors; nanowire; next generation; novel; novel therapeutics; programs; prototype; reaction rate; research and development; response; scale up; screening; sensor; success; tau Proteins; tau aggregation; tau-1; technology platform; technology validation; time use

Project Summary Global expenditures by pharmaceutical companies for research and development continue to increase each year with a current estimate of $1B USD and upwards of 15 years to develop a new drug. Drug discovery efforts are critical to the downstream success of candidate selection and relies heavily on high-throughput screening (HTS) to identify viable leads. Current methods for HTS drug discovery assays, such as fluorescence-, or luminescence-based methods, suffer from inherent technical drawbacks and limitations that result in false results leading to failed programs. Additionally, the majority of current HTS methods are not amendable to ultra high- throughput and use end-point assays as opposed to real-time kinetic approaches that can provide a more thorough assessment of molecular interactions. The use of real-time kinetic assays and higher throughput in drug discovery programs could greatly enhance success rates while reducing cost and time. INanoBio is developing a novel fully depleted exponentially coupled (FDEC) field effect transistor (FET) biosensor-based nanosensor multiplexed electronic drug discovery platform (nMEDD) for HTS. Our nMEDD platform will offer (i) real time enzymatic reaction monitoring; (ii) ultra-high sensitivity of detection; (iii) ultra HTS scalability; and (iv) high automation compatibility. The FDEC FET sensors electronically monitor changes in charge or potential by directly reacting with ions in solution, thus allowing for the determination of kinetic reaction rates to inform real-time detection and quantitative measurement of the effects of inhibitors or activators on the enzymatic reaction. As an initial validation of the technology platform, we are focusing on developing nMEDD for use in drug discovery for Alzheimer’s disease (AD). Specifically, we will focus on developing assays to monitor tau phosphorylation and dephosphorylation to enable screening for modulators of tau phosphorylation, as cytosolic aggregates of tau have been linked to AD pathology. The overall goal of this Fast-Track program is to develop INanoBio’s nMEDD platform as a HTS method for monitoring enzymatic reactions for drug discovery purposes. To achieve this goal, the Phase I program will be focused on development of a small-scale multiplex sensor and FDEC FET assays for tau phosphorylation and dephosphorylation to validate the performance of the multiplex sensor. Successful completion of Phase I will result in a sensor chip with spacing that is applicable to a 1536-well plate format and is capable of providing a stable response across the chip for detection of tau phosphorylation and dephosphorylation. The Phase II program will focus on further demonstrating the commercial potential of the FDEC FET technology by completing fabrication of an FDEC FET multiplex sensor and detection system for 1536-well detection of phosphorylation and dephosphorylation of tau, in addition to validation of the assay and demonstration of its potential for higher throughput. Successful completion of the proposed program will provide validation of the nMEDD technology for drug discovery to support partnering efforts and inform expansion of the technology into other disease areas.

项目概要 全球制药公司的研发支出持续增加 目前估计需要 10 亿美元，需要 15 年以上的时间来开发一种新药。药物发现工作 对于下游候选人选择的成功至关重要，并且在很大程度上依赖于高通量筛选 （HTS）以确定可行的线索。当前用于 HTS 药物发现测定的方法，例如荧光或 基于发光的方法存在固有的技术缺陷和限制，导致错误的结果 导致程序失败。此外，大多数当前的高温超导方法都不能修改为超高 吞吐量并使用终点检测，而不是实时动力学方法，可以提供更多 分子相互作用的彻底评估。使用实时动力学分析和更高的通量 药物发现计划可以大大提高成功率，同时降低成本和时间。 INanoBio 正在开发一种新型全耗尽指数耦合 (FDEC) 场效应晶体管 (FET) 用于 HTS 的基于生物传感器的纳米传感器多重电子药物发现平台 (nMEDD)。我们的 nMEDD 平台将提供（i）实时酶反应监测； (ii) 超高的检测灵敏度； (iii) 超高温超导 可扩展性； (iv) 高自动化兼容性。 FDEC FET 传感器以电子方式监控 通过直接与溶液中的离子反应来测量电荷或电势，从而可以测定动力学反应 比率以通知实时检测和定量测量抑制剂或激活剂对 酶促反应。作为技术平台的初步验证，我们专注于开发 nMEDD 用于治疗阿尔茨海默病（AD）的药物发现。具体来说，我们将专注于开发监测方法 tau 磷酸化和去磷酸化，以筛选 tau 磷酸化调节剂，如 tau 细胞质聚集体与 AD 病理有关。 该快速通道计划的总体目标是开发 INanoBio 的 nMEDD 平台作为 HTS 方法，用于 监测酶反应以用于药物发现目的。为了实现这一目标，第一阶段计划将 专注于开发小型多重传感器和 FDEC FET 测定法，用于 tau 磷酸化和 去磷酸化以验证多重传感器的性能。一期工程顺利完成 导致传感器芯片的间距适用于 1536 孔板格式，并且能够提供 在整个芯片上具有稳定的响应，用于检测 tau 磷酸化和去磷酸化。第二阶段 计划将重点通过完成进一步展示 FDEC FET 技术的商业潜力 用于 1536 孔磷酸化检测的 FDEC FET 多重传感器和检测系统的制造 和 tau 去磷酸化，此外还验证了该测定并证明了其更高的潜力 吞吐量。拟议计划的成功完成将为 nMEDD 技术提供验证 药物发现，以支持合作努力并为该技术扩展到其他疾病领域提供信息。