Identify small molecule inhibitors of methyl-dependent protein-protein interactio

描述（由申请人提供）：随着我们对表观遗传学理解的扩大，越来越清楚的是，这一领域的参与者是药物开发的潜在目标。表观遗传疗法的这一领域已经建立了治疗癌症和神经系统疾病的应用。我们已经在这一领域工作了多年，并进行了高通量筛选，以确定第一个小分子蛋白质甲基转移酶抑制剂。这些甲基转移酶靶向多种底物，从而调节表观遗传途径的许多环节。不同底物的甲基化通常会产生含有结构域（chromo、tudor、PHD、MBT和ANK重复序列）的效应蛋白的对接位点。我们假设，通过抑制特定的蛋白质-蛋白质相互作用，我们将能够开发出阻断表观遗传途径单臂的化合物。阻断单个蛋白质-蛋白质相互作用所获得的特异性将远远大于阻断调节多种相互作用之一的酶所获得的特异性。低亲和蛋白-蛋白相互作用（Kd在一到两位数的微摩尔范围内），如这里讨论的，很难用传统的技术，如荧光偏振或荧光共振能量转移来测定。我们通过开发一种敏感的基于化学发光的分析方法解决了这个问题，这是本项目的起点。在这里，我们计划进行筛选，以确定可以阻断甲基依赖的蛋白质-蛋白质相互作用的先导化合物，这可以用作分析这些甲基驱动的相互作用的探针，甚至可能发展成靶向表观遗传疗法。)

DESCRIPTION (provided by applicant): As our understanding of epigenetics expands, it is becoming clear that players in this field are potential targets for drug development. This area of epigenetic therapy has established applications for the treatment of cancer and neurological diseases. We have been involved in this field for a number of years and have performed high-throughput screens to identify the first small molecule inhibitors of protein methyltransferases. These methyltransferases target multiple substrates, and thus regulate many arms of an epigenetic pathway. The methylation of different substrates often generates docking sites for effector protein that harbor domains (chromo, tudor, PHD, MBT and ANK repeats). We hypothesize that by inhibiting specific protein-protein interactions we will be able to develop compounds that block a single arm of an epigenetic pathway. The specificity attained by blocking a single protein-protein interaction will be far greater than that attained by blocking the enzyme that regulates one of many interactions. Low-affinity protein-protein interactions (Kd in the single- to double-digit micromolar range) such as those discussed here have been very difficult to assay by traditional techniques such as fluorescence polarization or fluorescence resonance energy transfer. We have solved this problem by developing a sensitive chemiluminescence-based assay, which serves as the starting point for this project. Here we plan to perform screens to identify lead compounds that can block methyl- dependent protein-protein interactions, which can be used as probes to analyze these methyl-driven interactions and perhaps even developed into targeted epigenetic therapies. ) PUBLIC HEALTH RELEVANCE: Epimutations, unlike genetic mutations, can be reversed by chemotherapeutic intervention, which makes epigenetic therapy conceptually extremely appealing. Drug therapies that target chromatin have been touted as the next emerging frontier for the treatment of cancer and neuronal-based diseases. In this study, we will identify small molecules that will prevent these epimutations from being read.