Development of preclinical HTS for discovery of drugs modulating apoptosis

目前的临床前高通量药物筛选方法耗时且昂贵。此外，委员会认为， 在筛选过程中产生的大多数候选人在进一步的动物试验后被证明是无效的 模型为了克服这些局限性，现在正在努力开发非侵入性，高度敏感的 以及用于监测和早期检测单细胞和体内药物功效的成本有效的成像工具。 可激活的成像探针被设计成响应于识别而放大或增强荧光信号 特定的生物分子相互作用。我们开发了各种蛋白酶激活的成像系统， 通过使用具有荧光团，肽， 聚合物和金属。在这项提案中，我们计划将这些研究成果扩展到一种新的临床前药物 筛选系统，用于快速表征与糖尿病相关的候选药物。由于大多数 有效的抗癌治疗剂介导的凋亡级联，真实的时间筛选方法来检测 在体外和体内的单个细胞中，细胞凋亡在分子水平上的进展将显著改善 细胞内凋亡信号机制的理解，可以帮助临床监测， 筛查糖尿病相关药物的疗效。我们将开发先进的筛选技术， 应用于基于细胞的高通量和体内药物筛选，用于下一代药物靶向 凋亡不同类型的可激活成像系统靶向关键的凋亡介质，半胱天冬酶， 将被开发和优化用于体外和体内应用。通过靶向一个关键的细胞凋亡 通过介导，细胞凋亡的早期阶段可以容易地以快速和快速的方式被监测、成像和分析。 高效时尚选定的筛选系统将在各种肿瘤实验模型中进行验证 凋亡将起草标准的体外和体内药物筛选方法。总的来说，这个系统是 预计将大大减少与动物药物筛选试验相关的时间和成本。我们的目标是 开发完全可翻译的临床前筛选系统，用于靶向细胞凋亡的药物发现。成功 在这一倡议中说明的概念验证将允许广泛的临床前和临床应用。

Current preclinical high-throughput drug screening methods are time-consuming and expensive. Moreover, most of the candidates generated during screenings turn out to be invalid after further testing in animal models. To overcome these limitations, efforts are now being made to develop noninvasive, highly sensitive and cost-effective imaging tools for monitoring and early detection of drug efficacy in single cells and in vivo. Activatable imaging probes are designed to amplify or boost fluorescence signals in response to recognition of specific biomolecular interactions. We developed various protease-activatable systems for imaging and diagnostic applications by using advanced nanobioconjugation chemistry with fluorophores, peptides, polymers and metals. In this proposal, we plan to expand those research findings to a novel preclinical drug screening system for the rapid characterization of apoptosis-related drug candidates. Because the most effective anticancer therapeutics mediate apoptosis cascades, real time screening methods to detect the progression of apoptosis at the molecular level in single cells in vitro and in vivo would significantly improve the understanding of intracellular apoptotic signal mechanisms and could assist the clinical monitoring and screening of apoptosis-related drug efficacy. We will develop advanced screening technologies that can be applied in cell-based high-throughput and in vivo drug screening for next generation drugs targeting apoptosis. Different types of activatable imaging systems that target crucial apoptosis mediators, caspases, will be developed and optimized for in vitro and in vivo applications. By targeting a crucial apoptosis mediator, the early stage of apoptosis can be readily monitored, imaged and analyzed in a rapid and efficient fashion. Selected screening systems will be validated in various experimental models of tumor apoptosis. Standard in vitro and in vivo drug screening methods will be drafted. Overall, this system is expected to significantly reduce time and costs associated with drug screening tests in animals. Our aim is to develop fully translatable preclinical screening systems for drug discovery targeting apoptosis. Success of the proof-of-concept illustrated in this initiative will allow for widespread preclinical and clinical applications.