Fluorescence Lifetime-Based Kinase Assay for HTS

描述(由申请人提供)：基于荧光的分析极大地促进了药物发现中的高通量筛选(HTS)。然而，在这样做的过程中，他们造成了与假阳性和假阴性相关的瓶颈，这可能是由于光散射、内滤波效应和复合荧光的干扰而产生的。与荧光强度不同，基于荧光寿命的测量抗光散射和内滤波效应，并提供区分目标荧光信号和背景荧光所需的选择性。在第一阶段，我们将演示一种均一的基于荧光寿命的激酶分析方法，该方法可以抵抗荧光干扰，并且不需要使用抗体。克服荧光干扰将减少与二次筛选相关的成本，并有助于更快地鉴定先导化合物。此外，它将创造机会，筛选已知具有某些荧光特性，但以前无法通过传统的基于荧光的方法获得的化合物文库和/或天然产品。激酶是一类主要的药物靶点，与癌症、糖尿病和关节炎等人类疾病有关。这项技术将提高我们找到治疗这些重要疾病的药物的速度和效率。

DESCRIPTION (provided by applicant): Fluorescence based-assays have greatly facilitated high-throughput screening (HTS) in drug discovery. However, in doing so, they created a bottleneck relating to false positives and false negatives, which may arise due to interferences from light scattering, inner filter effects, and compound fluorescence. Unlike fluorescence intensity, a fluorescence lifetime-based measurement is resistant to light scattering and inner filter effects, and provides the selectivity needed to discriminate the target fluorescence signal from background fluorescence. In Phase I we will demonstrate a homogenous fluorescence lifetime-based kinase assay that is resistant to fluorescence interferences and does not require the use of antibodies. Overcoming fluorescence interferences will cut costs associated with secondary screening and assist quicker identification of lead compounds. Furthermore, it will create an opportunity to screen compound libraries and/or natural products that are known to have some fluorescent properties, but were previously inaccessible by means of traditional fluorescence-based methods. Kinases are a major class of drug targets, which are involved in human diseases such as cancer, diabetes, and arthritis. This technology will advance the speed and efficiency in which we can find drugs to treat these important diseases.