Discovery of Selective MAP2K4 Inhibitors to Target Metastasis

 描述(申请人提供)：转移性前列腺癌是不可治愈的，死亡主要是由于远处转移的发展造成的。同样，在所有癌症类型中，超过80%的死亡源于转移的形成，因此有效的抗转移治疗将具有巨大的临床益处。不幸的是，治疗方法一直受到缺乏针对调节转移的潜在分子过程的有效和选择性分子的限制。通过对小鼠和人的遗传学和化学方法的研究，我们发现MAP2K4是肿瘤转移的关键调控因子。这项提案的目标是识别和表征选择性MAP2K4抑制剂，这些抑制剂将作为探针，验证该激酶在一系列侵袭性癌症疾病模型中的作用。在前期工作中，我们开发了HTS的功能激酶分析(Z‘0.58)，并进行了中试筛选，确认了我们识别MAP2K4抑制剂的能力(命中率为1.1%)。我们还开发了正交生化和细胞分析来表征HTS HITS的选择性、机制和抗转移潜力。通过这项建议，我们将：1)筛选190,000种不同化合物的文库，以确定功能性MAP2K4抑制剂；2)使用生化分析确认MAP2K4的抑制和选择性，并通过模拟合成验证有前景的HIT支架；以及3)评估HITS的细胞活性，以确定效力、选择性和抗转移特性。我们预测，我们的综合方法将使新的MAP2K4抑制剂的发现成为可能，这将为生物界提供工具来评估转移在一系列疾病模型中的作用。此外，这些分子将作为未来的先导优化研究的起点，以创建抗转移临床候选药物。

 DESCRIPTION (provided by applicant): Metastatic prostate cancer is not curable, and mortality primarily results from the development of distant metastases. Similarly, over 80% of deaths across all cancer types stem from the formation of metastases, and therefore effective anti-metastatic treatments would have enormous clinical benefit. Unfortunately, therapeutic approaches have been limited by a lack of potent and selective molecules targeting the underlying molecular processes that regulate metastasis. Using genetic and chemical methods in mouse and man, we have identified MAP2K4 as a key regulator of metastasis. The goal of this proposal is to identify and characterize selective MAP2K4 inhibitors that would serve as probes to validate the role of this kinase in an array of invasive cancer disease models. In preliminary work, we developed a functional kinase assay for HTS (Z' 0.58) and performed a pilot screen confirming our ability to identify MAP2K4 inhibitors (1.1% hit rate). We have also developed orthogonal biochemical and cellular assays to characterize the selectivity, mechanism, and anti-metastatic potential of HTS hits. Through this proposal we will: 1) Screen a library of 190,000 diverse compounds to identify functional MAP2K4 inhibitors; 2) Confirm MAP2K4 inhibition and selectivity using biochemical assays and validate promising hit scaffolds through analog synthesis; and 3) evaluate cellular activity of hits to determine potency, selectivity, and anti-metastatic properties. We predict that our integrative approach will enable discovery of new MAP2K4 inhibitors, which will provide the biological community with tools to evaluate the role of metastasis in a range of disease models. Furthermore, such molecules will serve as a starting point in future lead optimization studies to create anti- metastatic clinical candidates.