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

基本信息

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
项目类别：
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

项目摘要

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，建议通过其专有的“转染酶和代谢芯片（或Teamchip）对化学安全技术的关键需求”（或Teamchip），以对系统药物候选药物和化学代理和毒理学和毒理学和毒理学和毒理学和毒理学和毒理学进行高通量分析。为模仿人肝脏的首次代谢，并预测酶特异性的肝毒性。代谢对其他细胞类型的影响也将得到证明。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。该阶段ISTTR项目的具体目的是：1。开发有效的方法将基因转染到THLE-2人肝上皮细胞系和BEAS-2B人类肺上皮细胞系中，该细胞封装在3D藻酸盐基质中，小至30 nl。 2。构建重组腺病毒，可携带来自人肝cDNA库中代谢酶的基因（例如CYP450的同工型，包括CYP1A2和CYP3A4），并在96-Well Plate中使用Fluororgenic substrate的THLE-2和Beas-2b细胞的单层转染了基因转染，并在96-Well Plate中进行了Beas-2b细胞。 3。演示3D细胞微阵列，其中包含表达THLE-2和BEAS-2B细胞的代谢酶，并鉴定出代谢基因的代谢基因，其差异表达会影响细胞对化学物质的细胞反应作为概念证明。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。 1 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I阶段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提出以进行铅优化以及更好和更安全的药物的最终开发。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。