Fluorescent HTS Assays for Human Sulfotransferases

描述（申请人提供）：药物代谢问题，如药物相互作用和毒性代谢产物的产生，导致临床试验期间几乎一半的候选药物失败。硫酸化是一种重要的共轭药物代谢类型，也用于调节内源性激素的水平和活性。例如，硫酸化调节特定神经递质受体、核受体和蛋白激酶的内源性配体的活性，这些受体和蛋白激酶是抑郁症、乳腺癌和心血管疾病的药物靶点。目前，由于缺乏高通量筛选试验来描述11种已知的人磺基转移酶对内源性激素和候选药物的特异性，制药公司对这些疾病的更具选择性的治疗方法的开发受到阻碍。为了解决这一需求，我们建议在第一阶段，以雌激素磺基转移酶（SULT1E1）为模型，提供基于荧光的磺基转移酶均相测定方法的原理证明。在第二阶段，新的测定方法将进行优化，一个完整的面板的11人磺基转移酶将被纯化和纳入，并通过筛选不同的生物活性化学品库，用于识别SULT底物和抑制剂，该测定将被验证用于药物HTS应用。

DESCRIPTION (provided by applicant): Drug metabolism problems such as drug-drug interactions and production of toxic metabolites cause almost half of all drug candidate failures during clinical trials. Sulfation is an important type of conjugative drug metabolism that is also used to regulate the level and activity of endogenous hormones. For instance, sulfation regulates the activity of endogenous ligands for specific neurotransmitter receptors, nuclear receptors, and protein kinases that are drug targets for depression, breast cancer, and cardiovascular disease. Development of more selective therapies for these disorders by pharmaceutical companies is currently hampered a lack of high throughput screening assays for delineating the specificity of the 11 known human sulfotransferases for endogenous hormones and drug candidates. To address this need, we propose in Phase I to provide proof of principle for a fluorescence based homogenous assay method for sulfotransferases using estrogen sulfotransferase (SULT1E1) as a model. In Phase II, the novel assay methods will be optimized, a full panel of 11 human sulfotransferases will be purified and incorporated, and the assay will be validated for pharmaceutical HTS applications by screening a diverse library of bioactive chemicals for identification of SULT substrates and inhibitors.