Identification and characterization of selective azaphilone inhibitors of HuR-mRNA interactions

建议书摘要 RNA结合蛋白通过与mRNAs结合来调节基因表达，从而影响基因表达的速率。 翻译、信使核糖核酸亚细胞定位和信使核糖核酸半衰期。RBP-mRNAs的特异性功能产物 相互作用取决于两个绑定伙伴的身份。然而，要预测 由于RBP-mRNA相互作用的不同组合而导致的特定相互作用的功能后果 发生在细胞内。RBP Hur结合有许多不同的mRNA结合伙伴，允许HUR 来控制许多关键的细胞功能。HUR-mRNA相互作用网络的失调是值得注意的 与结肠癌、肺癌和胰腺癌等癌症有关，这是美国最致命的三种癌症 各州。最近发现的一种氮杂环酮HUR抑制剂非常有效，但缺乏所需的选择性 治疗发展或解剖复杂的Hur mRNA结合伙伴网络。传统合成 方法是对映体选择性氮卓酮合成的主要障碍，使得筛选变得不可行。 类似物具有更好的生物活性。然而，最近的创新产生了直截了当的、一刀切的 生物催化路线，可用于产生大型、多样化的氮杂环丙酮库。我的目标是识别小说 氮杂环丙酮类作为有效和选择性的Hur-mRNA抑制剂，使用组合的生物催化生成和 HUR结合分析平台。我将表征和验证Hur-azapilone的相互作用，并确定选择性 氮卓酮在体外和细胞内对HUR的抑制作用。接下来，我将调查中断的功能影响 癌细胞模型中特定的hur-mrna相互作用。拟议工作的完成将导致 高效、选择性Hur-mRNA相互作用的新型氮杂环酮类化合物的鉴定与鉴定 并可作为化学探针和癌症治疗药物进一步开发。

PROPOSAL SUMMARY RNA-binding proteins (RBPs) regulate gene expression through binding to mRNAs, thus influencing rates of translation, mRNA subcellular location, and mRNA half-life. The specific functional outcome of an RBP-mRNA interaction is dependent on the identity of both binding partners. However, it is challenging to predict the functional consequences of a specific interaction due to the diverse combinations of RBP-mRNA interactions that occur within the cell. The RBP HuR binds has a multitude of different mRNA binding partners, allowing HuR to control many critical cellular functions. Dysregulation of the HuR-mRNA interaction network is notably implicated in cancers such as colon, lung, and pancreatic cancers, the three most deadly cancers in the United States. A recently discovered azaphilone HuR inhibitor is very potent but lacks the selectivity required for therapeutic development or to dissect the complex network of HuR mRNA binding partners. Traditional synthetic methods are a major roadblock towards enantioselective azaphilone synthesis, making it infeasible to screen analogs for more favorable bioactivity. However, recent innovation has produced a straightforward, one-pot biocatalytic route that can be used to generate large, diverse azaphilone libraries. I aim to identify novel azaphilones that act as potent and selective HuR-mRNA inhibitors using a combined biocatalytic generation and HuR binding assay platform. I will characterize and validate HuR-azaphilone interactions and identify selective azaphilone inhibitors of HuR both in vitro and in cellulo. Next, I will investigate functional implications of disrupting specific HuR-mRNA interactions in a cancer cell model. Completion of the proposed work will result in the identification and characterization of novel azaphilones that are potent and selective HuR-mRNA interactions and can be further developed as chemical probes and cancer therapeutics.