TeamChip for High-throughput, Predictive Human Metabolism and Toxicology

TeamChip 用于高通量、预测性人体代谢和毒理学

• 批准号: 7803769
• 负责人: MOO-YEAL LEE
• 金额: $ 15万
• 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 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7803769
• 关键词: Acetaminophen; Address; Adenoviruses; Affect; Alginates; Animals; Biological Assay; CYP1A2 gene; CYP3A4 gene; Catalysis; Cell Line; Cells; Chemicals; Clinical Trials; Cyclophosphamide; Cytochrome P450; Data; Data Analyses; Development; Drug or Chemical; Encapsulated; Environmental Pollutants; Enzymes; Epithelial Cells; Evaluation; Exposure to; Fluorogenic Substrate; Generations; Genes; Glass; Green Fluorescent Proteins; Hepatocyte; Hepatotoxicity; Human; Human body; In Vitro; Individual; Institutes; Investments; Lead; Libraries; Liver; Lung; Measurement; Metabolic; Metabolism; Methods; Organ; Parents; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Population; Predisposition; Process; Protein Isoforms; Protocols documentation; Public Health; Recombinants; Reporter Genes; Research; Research Personnel; Safety; Screening procedure; Simulate; Slide; Small Business Technology Transfer Research; Speed; Staging; Subgroup; System; Technology; Testing; Therapeutic; Time; Toxic effect; Toxicology; Transfection; Treatment Protocols; Viral; Xenobiotic Metabolism; Xenobiotics; base; cDNA Library; cell type; design; drug candidate; drug development; drug discovery; environmental chemical; flexibility; high risk; high throughput analysis; in vivo; monolayer; programs; public health relevance; red fluorescent protein; response; tool; toxicant

DESCRIPTION (provided by applicant): Solidus Biosciences, Inc. in partnership with Rensselaer Polytechnic Institute is proposing to address a critical need in chemical safety technology through 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 the first-pass metabolism of the human liver and to predict enzyme-specific hepatotoxicity. The effects of metabolism on other cell types will also be demonstrated. 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 of this Phase I STTR project are to: 1. Develop efficient methods for transfecting genes into THLE-2 human liver epithelial cell lines and Beas-2B human lung epithelial cell lines encapsulated in 3D alginate matrices as small as 30 nL. 2. Construct recombinant adenoviruses that carry genes for metabolic enzymes from a human liver cDNA library (e.g., CYP450 isoforms including CYP1A2 and CYP3A4) and demonstrate gene transfection on monolayers of THLE-2 and Beas-2B cells using fluorogenic substrates in a 96-well plate. 3. Demonstrate 3D cellular microarrays containing metabolic enzyme-expressing THLE-2 and Beas-2B cells and identify metabolic genes whose differential expression affects the cellular response to chemicals as proof of concept. 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. 1 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 I STTR project for the development of Solidus Bioscience's TeamChip technology has significant relevance to public health by providing pharmaceutical researchers with the 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，提议通过其专有的“转染酶和代谢芯片”（或TeamChip）来解决化学安全技术的关键需求，用于系统候选药物和化学代谢和毒理学的高通量分析。开发TeamChip是为了模拟人类肝脏的第一次代谢，并预测酶特异性肝毒性。新陈代谢对其他细胞类型的影响也将被证明。因此，目标化合物与人类个体代谢酶或人类肝脏或其他器官中酶的组合的反应性可以以与早期候选药物和环境化学物质的预测性人体毒性评估相称的速度进行评估和量化。第一阶段str项目的具体目标是：建立有效的方法将基因转染到小至30 nL的三维海藻酸盐基质中包裹的THLE-2人肝上皮细胞系和Beas-2B人肺上皮细胞系。2. 构建携带人肝脏cDNA文库中代谢酶基因的重组腺病毒（例如，CYP450亚型，包括CYP1A2和CYP3A4），并在96孔板上使用荧光底物对THLE-2和Beas-2B细胞单层进行基因转染。3. 展示包含代谢酶表达THLE-2和Beas-2B细胞的3D细胞微阵列，并鉴定代谢基因，其差异表达影响细胞对化学物质的反应，作为概念证明。可用于快速评估大量化合物毒性的体外技术仍然有限。安全评价的一个关键组成部分是化学品（例如，候选药物和环境化学毒物）的代谢和毒理学，这反映了化学品被人体代谢酶代谢的敏感性以及母体化合物及其代谢物的毒性。目前的化学品安全评估方法成本高，耗时长，使用大量的化合物和大量的动物。因此，将TeamChip作为一种安全评估工具，用于评估特定代谢酶是否以及如何影响候选药物和化学毒物的毒性，具有巨大的潜力和机会。这种能力也可用于预测个体在药物和化学代谢和毒性方面的差异。1