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: 10450925
• 负责人: Bharath Takulapalli
• 金额: $ 97.47万
• 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/10450925
• 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; 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; Play; Protein Dephosphorylation; Publishing; Reaction; Reporting; Reproducibility; Role; Scanning; Semiconductors; Signal Transduction; Surface; System; Technology; Testing; Time; Transistors; Validation; Variant; assay development; base; candidate selection; chemical reaction; cost; data acquisition; detection assay; detection platform; detection sensitivity; drug discovery; electronic sensor; enzyme activity; enzyme substrate; high throughput screening; high-throughput drug screening; inhibitor; kinase inhibitor; luminescence; multiplex detection; 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 phosphorylation; tau-protein kinase; 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.

项目总结