Development of a high-throughput screen to detect the effects of both pre- and post-biotransformed compounds for enhanced content drug discovery workflows

开发高通量筛选来检测生物转化前和生物转化后化合物的影响，以增强药物发现工作流程的内容

• 批准号: 9466709
• 负责人: GARY S SAYLER
• 金额: $ 52.26万
• 依托单位: 490 BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9466709
• 关键词: Address; Affect; Biological Assay; Biotechnology; CYP1A2 gene; CYP2B6 gene; CYP3A4 gene; Cardiac; Cell Culture System; Cell Culture Techniques; Cell Death; Cell Line; Cell model; Cells; Chemicals; Coculture Techniques; Complex; Data; Detection; Development; Dimensions; Drug Industry; Drug Metabolic Detoxication; EPHX1 gene; Early Diagnosis; Economics; Effectiveness; Epithelial; Evaluation; Expenditure; Failure; Fees; Generations; Genetic; Gold; Growth; Health; Human; Human Biology; Human body; Image; In Vitro; Industrialization; Industry; Kidney; Legal; Libraries; Light; Link; Liver; Logistics; Luciferases; Measurement; Metabolic; Metabolism; Methods; Modernization; NAT1 gene; National Institute of General Medical Sciences; Nature; Organ; Output; Pathway interactions; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase; Phase I Clinical Trials; Physiological; Physiology; Predisposition; Prodrugs; Production; Regimen; Reporter; Research Personnel; Rofecoxib; Safety; Sampling; Signal Transduction; Small Business Innovation Research Grant; System; Technology; Therapeutic; Time; Tissues; Toxic effect; Toxicity Tests; Toxicology; Vascular Endothelium; Work; assay development; base; cell type; cerivastatin; comparative; cost; cytotoxic; data acquisition; drug development; drug discovery; drug metabolism; established cell line; high throughput screening; improved; in vivo; injured; killings; novel; novel therapeutics; operation; pancreatic cell line; research and development; scaffold; screening; technology validation; three dimensional cell culture

Development of a high-throughput screen to detect the effects of both pre- and post- biotransformed compounds for enhanced content drug discovery workflows Project Summary This Small Business Innovation Research Phase II project will build upon our successful Phase I demonstration that substrate-free autobioluminescent signal generation can detect both the pre- and post-biotransformed metabolic impacts of therapeutic compounds from a single plate-based assay. Here, we will leverage this technology to develop a panel of industry-relevant autobioluminescent cell lines optimized for the detection of pre- and post-biotransformed compound metabolic impacts and the identification of specific detoxification pathway activation using modern three-dimensional (3D) microphysiological culture systems. These products and their underlying technology will specifically address the National Institute of General Medical Sciences (NIGMS) request for novel in vivo and in vitro methods for predicting the safety and toxicities of pharmacologic agents. By optimizing this technology to function within the industry-preferred 3D microphysiological format, we will address the critical need for new methods that can both identify compound toxicity and elucidate the mechanisms through which cells mitigate the compounds’ effects. The autonomous nature of this technology will increase toxicological data acquisition while preserving the critical advantage of presenting physiologically- relevant data, and reducing the cost of performance by eliminating substrates, reducing complexity, limiting hands-on operation time, obviating the need for sample destruction, and reducing the potential for measurement error. Through the validation of this technology at a scale relevant to tier 1 drug discovery screening and its comparative analysis against the existing gold-standard ATP content assay, this revolutionary approach is poised to have a significant and immediate impact towards reducing the estimated $8B/year in unnecessary expenditures made by pharmaceutical companies during their development of the 48% of new compounds that fail at the Phase I clinical trial stage due to misidentification of toxicological effects during tier 1 screening. This is possible because, as demonstrated in our Phase I work, the use of our autobioluminescent technology overcomes the high economic and logistical costs of existing, traditionally-bioluminescent cell’s requisite chemical substrate addition, which must co-occur with each generation of signal, and the intensive hands-on time necessitated to scale cultures due to their requisite sample destruction concurrent with imaging. Similarly, our autobioluminescent technology also obviates the hurdles presented by fluorescent cell’s susceptibility to autofluorescent signal inhibition and their tendency to remain active during downturns in cellular metabolism or even after cell death. The technology and products developed in this effort will therefore be capable of significantly improving the throughput and effectiveness of microphysiological systems-based tier 1 compound screening to improve the efficiency and economics of new compound development, and ultimately, consumer safety. This will allow them to thrive in a microphysiological system market that is predicted to maintain a compound annual growth rate of 70% to exceed $1.3B globally by 2022.

一种高通量屏幕的开发，以检测治疗前后的影响 用于增强内容药物发现工作流程的生物转化化合物 项目摘要 此小型企业创新研究第二阶段项目将建立在我们成功的第一阶段演示的基础上 这种无底物的自体发光信号的产生可以检测到生物转化前后的 来自单一平板化验的治疗化合物的代谢影响。这里，我们将利用这一点 开发一组与行业相关的自体发光细胞系的技术，优化用于检测 生物转化前后化合物代谢的影响及特异性解毒的鉴定 使用现代三维(3D)微生理培养系统激活通路。这些产品 他们的基础技术将专门针对国家普通医学科学研究所 (NIGMS)要求建立新的体内和体外药物安全性和毒性预测方法 探员们。通过优化该技术以在行业优选的3D微生理格式内工作， 我们将解决对新方法的迫切需求，这些方法既能识别化合物毒性，又能阐明 细胞通过哪些机制减轻化合物的影响。这项技术的自主性将 增加毒物学数据采集，同时保留呈现生理学- 相关数据，并通过消除底物、降低复杂性、限制 动手操作时间，消除了样品销毁的需要，并减少了测量的可能性 错误。通过在与第一级药物发现筛选相关的规模上验证这项技术 与现有的黄金标准ATP含量测定进行比较分析，这一革命性的方法势在必行 对减少估计每年80亿美元的不必要开支产生重大而直接的影响 制药公司在开发48%的新化合物期间的支出 在第一阶段临床试验阶段，由于在第一级筛查期间对毒理学效应的错误识别而失败。这 这是因为，正如我们在第一阶段工作中所展示的那样，我们的自体生物发光技术的使用 克服了现有的、传统生物发光细胞所需的高昂的经济和物流成本 化学底物添加，这必须与每一代信号同时发生，以及密集的动手 由于其必要的样品销毁与成像同时进行，因此扩大培养所需的时间。同样， 我们的自体生物发光技术还消除了荧光细胞对 自体荧光信号抑制及其在细胞代谢低迷时保持活跃的趋势 甚至在细胞死亡之后。因此，在这一努力中开发的技术和产品将能够 显著提高基于微生理系统的一级化合物的吞吐量和有效性 筛选以提高新化合物开发的效率和经济性，并最终提高消费者 安全。这将使他们在微生理系统市场中蓬勃发展，预计该市场将保持 到2022年，全球的复合年增长率将超过13亿美元。