Discovery of Selective MAP2K4 Inhibitors to Target Metastasis

 描述（由申请人提供）：转移性前列腺癌是不可治愈的，死亡率主要由远处转移的发展引起。类似地，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，治疗方法受到缺乏靶向调节转移的潜在分子过程的有效和选择性分子的限制。在小鼠和人类中使用遗传和化学方法，我们已经确定MAP 2K 4是转移的关键调节因子。该提案的目标是鉴定和表征选择性MAP 2K 4抑制剂，其将作为探针来验证该激酶在一系列侵袭性癌症疾病模型中的作用。在初步工作中，我们开发了HTS的功能性激酶测定（Z' 0.58），并进行了初步筛选，证实了我们鉴定MAP 2K 4抑制剂的能力（1.1%命中率）。我们还开发了正交生物化学和细胞测定来表征HTS命中的选择性、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鉴定功能性MAP 2K 4抑制剂; 2）使用生物化学测定确认MAP 2K 4抑制和选择性，并通过类似物合成验证有希望的命中支架;和3）评估命中的细胞活性以确定效力、选择性和抗转移性质。我们预测，我们的综合方法将能够发现新的MAP 2K 4抑制剂，这将为生物界提供评估转移在一系列疾病模型中的作用的工具。此外，此类分子将作为未来先导物优化研究的起点，以产生抗转移的临床候选物。

 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.