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既耗时又昂贵。此外， 在进一步的动物试验中，在筛选过程中产生的大部分候选被证明是无效的 模特们。为了克服这些限制，现在正在努力开发非侵入性的、高度敏感的 以及经济高效的成像工具，用于监测和早期检测单细胞和体内的药物疗效。 可激活的成像探针被设计为放大或增强荧光信号以响应识别 特定的生物分子相互作用。我们开发了各种可激活蛋白酶的系统，用于成像和 使用先进的纳米生物偶联化学与荧光团、多肽、 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 用于快速鉴定细胞凋亡相关候选药物的筛选系统。因为最多的 有效的抗癌治疗药物介导细胞凋亡级联反应，实时筛选方法检测 单细胞在体内外分子水平上的凋亡研究进展将显著改善 了解细胞内凋亡信号的机制，有助于临床监测和治疗 细胞凋亡相关药物疗效的筛选。我们将开发先进的筛查技术，可以 用于下一代药物靶向的细胞高通量体内药物筛选 细胞凋亡。不同类型的可激活的成像系统，针对关键的细胞凋亡介质，半胱氨酸氨基转移酶， 将被开发和优化用于体外和体内应用。通过靶向一种关键的细胞凋亡 中介物，可以很容易地监测，成像和分析细胞凋亡的早期阶段 高效的时尚。选定的筛查系统将在各种肿瘤实验模型中得到验证 细胞凋亡。将起草标准的体外和体内药物筛选方法。总体而言，这个系统是 预计将显著减少与动物药物筛选测试相关的时间和成本。我们的目标是 开发完全可翻译的临床前筛选系统，用于针对细胞凋亡的药物发现。取得的成功 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.