TeamChip for High-throughput, Predictive Human Metabolism and Toxicology

描述（由申请人提供）：Solidus Biosciences，Inc.该公司与伦斯勒理工学院合作，提出通过其专有的“转染酶和代谢芯片”（或TeamChip）来解决化学安全技术的关键需求，用于系统性候选药物和化学代谢及毒理学的高通量分析。正在开发的TeamChip模拟人类肝脏的首过代谢，并预测酶特异性肝毒性。代谢对其他细胞类型的影响也将得到证明。因此，可以以与早期候选药物和环境化学品的预测性人体毒性评估相当的速度评估和定量目标化合物与人体肝脏或其他器官类型中的单个人体代谢酶或酶组合的反应性。 第一阶段STTR项目的具体目标是：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人肺上皮细胞系中，这些细胞系封装在小至30 nL的3D藻酸盐基质中。2.构建携带来自人肝cDNA文库的代谢酶基因的重组腺病毒（例如，CYP 450同种型，包括CYP 1A 2和CYP 3A 4），并证明在96孔板中使用荧光底物在THLE-2和Beas-2B细胞单层上的基因转染。3.展示含有代谢酶表达THLE-2和Beas-2B细胞的3D细胞微阵列，并鉴定其差异表达影响细胞对化学品反应的代谢基因，作为概念验证。 可用于快速评估大量化合物毒性的体外技术仍然有限。安全性评价的一个关键组成部分是化学品的代谢和毒理学（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，并且使用大量化合物和大量动物。因此，有很大的潜力和机会，将TeamChip作为一种安全性评估工具，可用于评估特定的代谢酶是否以及如何影响候选药物和化学毒物的毒性。这种能力也可用于预测个体之间在药物和化学代谢和毒性方面的差异。1 公共卫生相关性：药物发现过程是一个投资密集型，高风险的奋进，导致低产量的有效和安全的药物;一个问题，是混淆的显着缺乏信息，存在于预测代谢的候选药物的命运，一般来说，并在预测候选药物在人体中的反应。拟议的第一阶段STTR项目，用于开发Solidus Bioscience的TeamChip技术，通过为制药研究人员提供预测候选药物体内代谢所需的信息，从而有助于决定哪些化合物被提出用于先导优化和最终开发更好，更安全的药物，对公共卫生具有重要意义。此外，这项研究与工业和环境化学品在安全和使用方面的优先次序有关。

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.