Computational and experimental studies of targets of protein kinase inhibitors

拟议的研究将是对相互作用的实验和计算相结合的研究。 在蛋白激酶和小分子抑制剂之间。蛋白激酶与许多 疾病，因此是有吸引力的治疗目标。然而，在设计小尺寸的 激酶的分子抑制剂是如何开发专门针对感兴趣的激酶的抑制剂，同时 身体中的数百种其他激酶不受影响。为了实现这一目标， 需要了解控制抑制物-激酶相互作用的特异性，其中一种方法是 开发能够准确预测抑制物-激酶的相对亲和力的计算方法 复合体。拟议的研究计划旨在测试、扩展和改进这种计算方法 通过大量的实验研究相结合。具体目标包括： (1)具有-10蛋白激酶范围的市售小分子抑制剂的抑制常数 目标将被测量。这些研究将提供关于抑制剂-激酶相互作用的新的定量数据。 (2)用等温滴定法测定抑制剂-激酶络合物的结合热力学。 量热法。将使用结合自由能、热焓和熵的单独测量来 对抑制物--激酶结合相互作用的能量描述进行参数化。 (3)具有一系列不同蛋白激酶靶点的复合体中的抑制剂的晶体结构将是 解决了。这些研究将提供对约束程度的直接和全面的看法 当与不同的蛋白激酶络合时，一种特定的抑制剂所采用的取向会改变。 (4)从目标1-3获得的实验数据将用于开发和测试一种计算方法。 用于准确预测抑制剂与ALL-500人蛋白激酶的相对结合亲和力。 蛋白激酶与许多不同的癌症类型有关，越来越多地被认为是 治疗药物的重要靶点。这里提出的研究直接针对单个最伟大的 对激酶治疗追求的挑战：如何设计能够特异性地 抑制一种感兴趣的激酶。

The proposed research will be a combined experimental and computational study of the interactions between protein kinases and small molecule inhibitors. Protein kinases are implicated in a number of diseases, and are therefore attractive therapeutic targets. A major challenge however in designing small molecule inhibitors of kinases is how to develop inhibitors that specifically target a kinase of interest while leaving the hundreds of other kinases in the body unaffected. In order to achieve this goal, the factors that control the specificity of inhibitor-kinase interactions need to be understood, and one way to do this is to develop computational methodologies that can accurately predict the relative affinities of inhibitor-kinase complexes. The proposed research plan aims to test, extend and refine such a computational methodology through combination with a number of experimental studies. The specific aims involve: (1) Inhibition constants of commercially available small molecule inhibitors with a range of -10 protein kinase targets will be measured. These studies will provide new quantitative data on inhibitor-kinase interactions. (2) The binding thermodynamics of inhibitor-kinase complexeswill be measured with isothermal titration calorimetry. Separate measurements of binding free energies, enthalpies and entropies will be used to parameterize an energetic description of inhibitor-kinase binding interactions. (3) The crystal structures of inhibitors in complex with a range of different protein kinase targets will be solved. These studies will provide a direct and comprehensive view of the extent to which the binding orientations adopted by a given inhibitor change when complexed with different protein kinases. (4) The experimental data obtained from Aims 1-3 will be used to develop and test a computational method for accurately predicting the relative binding affinities of an inhibitor with all -500 human protein kinases. Protein kinases, being implicated in a large number of different cancer types, are increasingly seen as very important targets for therapeutic drugs. The research proposed here directly addresses the single greatest challenge to the therapeutic pursuit of kinases: how to design small molecule inhibitors that can specifically inhibit a kinase of interest.