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

描述（由申请人提供）：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（或代谢酶稳定性测定芯片，或MesaChip）来解决药物安全技术的关键需求，用于高通量分析候选药物代谢。MesaChip的开发目的是为制药行业用户提供模拟人类肝脏首过代谢的能力。虽然近年来新化学实体（NCE）和可筛选药物靶点的数量急剧增加，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失败率很高。药物安全性评价的一个重要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易感性和代谢速率。然而，早期代谢稳定性分析目前受到固有的低通量方法的限制，这些方法可提供准确的候选药物代谢定量测量。因此，缺乏早期高质量决策所需的准确信息。这个第一阶段项目的具体目标是：1。通过同时测量氧气和NADPH的消耗以及H2O2的形成，开发出基于良好基础的荧光分析，以确定p450催化的底物氧化速率。2. 使用LC-MS验证上述速率测量可用于计算p450催化氧化反应的准确速率。3. 将Aim 1中开发的孔板检测技术应用于基于MetaChip平台的高通量微阵列格式（MesaChip），该格式涉及将p450整合到功能化玻璃载玻片上的微点（< 30 nL）中。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体内代谢稳定性所需的信息，从而帮助确定哪些化合物可以用于先导物优化。这种能力也是广泛采用个性化医疗的关键前兆，可以预测个体之间药物代谢的差异，从而在个体患者水平上提供药物治疗潜在结果的信息。因此，这个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，导致有效和安全药物的产量很低；由于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缺乏信息，使这个问题变得混乱。Solidus Bioscience公司MesaChip技术的STTR项目开发，为制药研究人员提供了预测候选药物体内代谢稳定性所需的信息，从而有助于确定哪些化合物被提出进行先导物优化，最终开发出更好、更安全的药物，与公共卫生具有重要的相关性。

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.