TeamChip for High-Throughput, Predictive Human Metabolism and Toxicology: Phase I

用于高通量、预测性人体代谢和毒理学的 TeamChip：第一阶段

• 批准号: 8251456
• 负责人: MOO-YEAL LEE
• 金额: $ 50万
• 依托单位: SOLIDUS BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8251456
• 关键词: Acetaminophen; Adenoviruses; Affect; Animals; Biological Assay; CYP2C9 gene; CYP3A4 gene; Cell Culture Techniques; Cells; ChIP-on-chip; Chemicals; Clinical Trials; Cryopreservation; Development; Encapsulated; Environmental Pollutants; Enzymes; Evaluation; Exposure to; Freezing; Generations; Genes; Hepatocyte; Hepatotoxicity; Human; Human body; In Vitro; Individual; Infection; Institutes; Investments; Lead; Libraries; Liver; Measurement; Measures; Metabolic; Metabolism; Methods; Modeling; Organ; Parents; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Plastics; Population; Predisposition; Process; Protein Isoforms; Proteins; Protocols documentation; Public Health; Recombinants; Reporter; Research; Research Personnel; Safety; Screening procedure; Simulate; Small Business Technology Transfer Research; Speed; Staging; Subgroup; System; Technology; Testing; Therapeutic; Time; Toxic effect; Toxicology; Transfection; Treatment Protocols; Validation; Viral; Western Blotting; Xenobiotic Metabolism; Xenobiotics; base; cDNA Library; cell type; commercialization; design; drug candidate; drug development; drug discovery; environmental chemical; flexibility; high risk; high throughput analysis; in vivo; monolayer; programs; response; tool; toxicant; troglitazone; Acetaminophen; Adenoviruses; Affect; Animals; Biological Assay; CYP2C9 gene; CYP3A4 gene; Cell Culture Techniques; Cells; ChIP-on-chip; Chemicals; Clinical Trials; Cryopreservation; Development; Encapsulated; Environmental Pollutants; Enzymes; Evaluation; Exposure to; Freezing; Generations; Genes; Hepatocyte; Hepatotoxicity; Human; Human body; In Vitro; Individual; Infection; Institutes; Investments; Lead; Libraries; Liver; Measurement; Measures; Metabolic; Metabolism; Methods; Modeling; Organ; Parents; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Plastics; Population; Predisposition; Process; Protein Isoforms; Proteins; Protocols documentation; Public Health; Recombinants; Reporter; Research; Research Personnel; Safety; Screening procedure; Simulate; Small Business Technology Transfer Research; Speed; Staging; Subgroup; System; Technology; Testing; Therapeutic; Time; Toxic effect; Toxicology; Transfection; Treatment Protocols; Validation; Viral; Western Blotting; Xenobiotic Metabolism; Xenobiotics; base; cDNA Library; cell type; commercialization; design; drug candidate; drug development; drug discovery; environmental chemical; flexibility; high risk; high throughput analysis; in vivo; monolayer; programs; response; tool; toxicant; troglitazone

DESCRIPTION (provided by applicant): Solidus Biosciences, Inc. in partnership with Rensselaer Polytechnic Institute, will focus on further development, validation, and commercialization of its proprietary "Transfected Enzyme and Metabolism Chip" (or TeamChip) for high-throughput analysis of systematic drug candidate and chemical metabolism and toxicology. The TeamChip is being developed to mimic first-pass metabolism of the human liver and to predict enzyme-specific hepatotoxicity. A library of human cells expressing different combinations of metabolic enzymes on the TeamChip will be prepared by transfecting metabolic genes using a viral delivery system into human cells encapsulated in three-dimensional (3D) matrices (as small as 60 nL) arrayed on a plastic chip. Thus, the reactivity of target compounds with individual human metabolic enzymes or combinations of enzymes in the human liver or other organ types can be assessed and quantified at speeds commensurate with predictive human toxicity assessment of early stage drug candidates and environmental chemicals. The specific aims/milestones of this Phase II STTR proposal are to: 1. Construct recombinant adenoviruses that carry genes for metabolic enzymes from a human liver cDNA library (e.g., representative CYP450 isoforms and conjugative metabolic enzymes), demonstrate gene transfection on monolayers of Hep3B cells using fluorogenic/luminescent substrates, and measure different levels of adenoviral enzyme expression by Western blot assays. 2. Prepare the TeamChip containing Hep3B cells expressing various combinations of metabolic enzymes and identify metabolic genes whose differential expression affects the cellular response to model compounds. 3. Optimize on-chip cryopreservation protocols for recombinant adenoviruses and transfected Hep3B cells. Demonstrate metabolism-induced toxicity with cryopreserved TeamChips and compare the results with non- frozen counterparts. In vitro technologies that can be used to quickly assess large numbers of compounds for toxicity remain limited. A critical component of safety evaluation is metabolism and toxicology of chemicals (e.g., drug candidates and environmental chemical toxicants), which reflects the susceptibility of chemicals to be metabolized by human metabolic enzymes and the toxicity of parent compounds and their metabolites. Current approaches to chemical safety assessment are costly, time consuming, and use large amounts of compound and large numbers of animals. Thus, there is great potential and opportunity to apply the TeamChip as a safety assessment tool that can be used to evaluate whether and how specific metabolic enzymes contribute to the toxicity of drug candidates and chemical toxicants. This capability may also be used to predict differences among individuals in drug and chemical metabolism and toxicity. PUBLIC HEALTH RELEVANCE: The drug discovery process is an investment-intensive, high-risk endeavor that results in low yields of effective and safe drugs; a problem that is confounded by the significant lack of information that exists in predicting the metabolic fate of drug candidates, in general, and in predicting the reactivity of drug candidates in the human body. The proposed Phase II STTR project for the development of Solidus Bioscience's TeamChip technology has significant relevance to public health by providing pharmaceutical researchers with in vitro information needed to predict the in vivo metabolism of drug candidates, and thus help to decide which compounds are brought forward for lead optimization and the ultimate development of better and safer drugs. Furthermore, this research is relevant to the prioritization of industrial and environmental chemicals in terms of their safety and use.

描述（由申请人提供）：Solidus Biosciences，Inc。与Rensselaer Polytechnic Institute合作，将重点介绍其专有的“转染酶和代谢芯片（或Teamchip）的进一步开发，验证和商业化”（或Teamchip），以对系统药物候选人和化学代理和质量学和化学代理和质量学和化学代理和化学代理和质量学和化学代谢分析。该团队正在开发以模仿人肝脏的第一频繁代谢，并预测酶特异性的肝毒性。通过使用病毒递送系统转染代谢基因将代谢酶在团队上表达不同组合的人类细胞库将制备，这些基因封装在塑料芯片上的三维（3D）基质中（小至60 nl）中的人类细胞。因此，可以评估和量化人肝脏或其他器官类型中酶的靶化合物的反应性或与早期药物候选药物和环境化学物质的预测性人类毒性评估的速度评估和量化。 该II期STTR建议的具体目的/里程碑是：1。构建重组腺病毒，这些腺病毒携带来自人肝cDNA文库的代谢酶的基因，例如，CYP450 CYP450同种型和共轭代谢酶的代表性酶，表现出对HexpoRent of Hexogenter of Hexogent of Hexogent of Hextrogent of Hextrogent of Hextricers of Hiphorent ofer cliprencation of Hextric inter的基因转移的不同。腺病毒酶通过蛋白质印迹测定法表达。 2.准备包含HEP3B细胞的TeamChip表达代谢酶的各种组合并鉴定代谢基因，其差异表达会影响模型化合物的细胞反应。 3.优化重组腺病毒和转染HEP3B细胞的芯片上冷冻保存方案。证明了与冷冻保存的团队一起新陈代谢诱导的毒性，并将结果与​​非冷冻对应物进行比较。 可用于快速评估大量毒性化合物的体外技术仍然有限。安全评估的关键组成部分是化学物质的代谢和毒理学（例如，候选药物和环境化学毒物），这反映了化学物质被人类代谢酶代谢的易感性以及父母代谢化合物的毒性及其代谢物的毒性。当前的化学安全评估方法是昂贵的，耗时的，并使用了大量的化合物和大量动物。因此，将团队策略应用于安全评估工具有很大的潜力和机会，可用于评估特定代谢酶是否以及如何促进候选药物和化学毒物的毒性。该能力也可以用于预测药物和化学代谢和毒性中个体之间的差异。 公共卫生相关性：药物发现过程是一项投资密集的，高风险的努力，可导致有效和安全的药物产量较低；通常，在预测候选药物的代谢命运以及预测人体中候选药物的反应性时，存在一个严重缺乏信息所困扰的问题。拟议的II期STTR项目用于开发Solidus Bioscience的TeamChip技术，通过为预测药物候选药物的体内代谢所需的体外信息提供了与公共卫生的重要相关性，从而有助于确定哪些化合物可以带来哪些化合物来提出以进行优化和更好和更安全的药物的最终发展。此外，这项研究与工业和环境化学物质在安全性和使用方面的优先级相关。