Fluorescent HTS Assays for Human Sulfotransferases

描述(由申请人提供)：在临床试验期间，几乎一半的候选药物失败是由于药物代谢问题，如药物与药物的相互作用和有毒代谢物的产生。硫酸盐化是一种重要的结合型药物代谢，也用于调节内源性激素的水平和活性。例如，硫酸盐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，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，因为缺乏高通量的筛选试验来描述11种已知的人内源性激素和候选药物的磺基转移酶的特异性。为了满足这一需要，我们在第一阶段提出了一种以雌激素磺基转移酶(SULT1E1)为模型的基于荧光的磺基转移酶均相分析方法的原理证明。在第二阶段，将优化新的检测方法，纯化并整合一整组11个人硫转移酶，并通过筛选用于鉴定底物和抑制剂的各种生物活性化学物库，验证该检测方法在药物HTS应用中的有效性。

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.