A High-Throughput Human Metabolic Stability Assay Chip for Early-Stage Drug Disco

描述(申请人提供)：Solidus Biosciences，Inc.与加州大学伯克利分校和伦斯勒理工学院合作，提议通过其专有的“代谢稳定性”芯片(或代谢酶稳定性分析芯片，或台式芯片)来满足药物安全技术的关键需求，用于高通量分析候选药物的新陈代谢。正在开发的台式芯片为制药行业的用户提供了模拟人类肝脏首次代谢的能力。虽然近年来新的化学实体(NCE)和可筛选药物靶标的数量急剧增加，但生产率的提高并没有转化为新药批准数量的增加，部分原因是NCE或其代谢物的毒性导致的高失败率(S)。药物安全性评价的一个关键组成部分是候选药物的代谢稳定性，它反映了候选药物被代谢的敏感性和代谢速度。然而，早期代谢稳定性分析目前受到现有方法固有的低吞吐量的限制，这些方法提供了对候选药物代谢的准确定量测量。因此，缺乏早期高质量决策所需的准确信息。这一第一阶段项目的具体目标是：1.开发基础良好的荧光分析方法，能够通过同时测量氧气和NADPH消耗以及过氧化氢的形成来确定P450催化的底物氧化速率。2.用LC-MS验证上述速率测量可用于计算P450催化的氧化反应的准确速率。3.使在目标1中开发的井板分析技术适用于基于MetaChip平台的高通量微阵列格式(台式芯片)，其涉及将P450整合到功能化玻璃片上的微斑点(&lt；30nl)。一旦完全开发，MesaChip将提供一个强大的高通量技术平台，能够为制药研究人员提供预测候选药物在体内代谢稳定性所需的信息，从而帮助决定提出哪些化合物进行先导优化。这一能力也是广泛采用个性化药物的关键先兆，可以预测个体在药物代谢方面的差异，从而提供关于个体患者层面药物治疗潜在结果的信息。因此，这一STTR项目的目标符合美国国立卫生研究院的目标。药物发现过程是一项投资密集、高风险的工作，导致有效和安全的药物产量较低；在预测候选药物的代谢命运和预测候选药物在人体内的反应性方面，普遍存在的严重缺乏信息使这一问题变得复杂起来。为开发Solidus Bioscience的中间芯片技术而拟议的STTR项目为制药研究人员提供了预测候选药物在体内的代谢稳定性所需的信息，从而有助于决定提出哪些化合物用于先导优化和最终开发更好、更安全的药物，因此对公众健康具有重要意义。

DESCRIPTION (provided by applicant): Solidus Biosciences, Inc. in partnership with the University of California, Berkeley, and Rensselaer Polytechnic Institute are proposing to address a critical need in drug safety technology through its proprietary "metabolic stability" chip (or Metabolizing Enzyme Stability Assay Chip, or MesaChip) for high-throughput analysis of drug candidate metabolism. The MesaChip is being developed to provide the pharmaceutical industry user with the ability to mimic the first-pass metabolism of the human liver. While in recent years there has been a dramatic increase in the number of new chemical entities (NCEs) and screenable drug targets, such increases in productivity have not translated into an increased number of new drug approvals, in part because of the high failure rate due to toxicity of the NCE or its metabolite(s). A critical component of drug safety evaluation is the metabolic stability of a drug candidate, which reflects the susceptibility of a drug candidate to be metabolized and the rate of such metabolism. Early stage metabolic stability analysis, however, is currently limited by the inherent low throughput of methods available that provide accurate quantitative measurement of drug candidate metabolism. As a result, accurate information required for early-stage high-quality decision making is lacking. The specific aims of this Phase I project are to: 1. Develop well-based fluorescence assays that enable determination of P450-catalyzed substrate oxidation rates from concurrent measurements of oxygen and NADPH consumption and of H2O2 formation. 2. Verify using LC-MS that the above rate measurements can be used to calculate accurate rates of P450-catalyzed oxidation reactions. 3. Adapt the well-plate assay techniques developed in Aim 1 to a high-throughput microarray format (MesaChip) based on the MetaChip platform that involves P450s incorporated into microspots (< 30 nL) on a functionalized glass slide. Once fully developed, the MesaChip will provide a robust and high-throughput technology platform capable of providing pharmaceutical researchers with the information needed to predict the in vivo metabolic stability of drug candidates, and thus help to decide which compounds are brought forward for lead optimization. This capability is also a critical precursor to the widespread adoption of personalized medicine, where differences among individuals in drug metabolism can be predicted, thereby providing information on the potential outcome of drug therapy at the individual patient level. The goals of this STTR project, therefore, fit the goals of the National Institutes of Health. 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 STTR project for the development of Solidus Bioscience's MesaChip technology has significant relevance to public health by providing pharmaceutical researchers with the information needed to predict the in vivo metabolic stability 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.