具有动力学选择性的c-Src抑制剂的设计、合成及其对多囊肾抑制活性研究

常染色体显性遗传性多囊肾病（ADPKD）发病率高、病程长且缺乏特效药。c-Src活性与ADPKD进程密切相关，是其重要的治疗靶标。现有的c-Src抑制剂因选择性低而带来严重的副作用，急需开发高选择性抑制剂。由于c-Src同家族的激酶域结构高度相似，基于传统热力学方法难以设计高选择性c-Src抑制剂。本项目聚焦c-Src抑制剂的动态解离过程，拟从动力学角度优化化合物与c-Src的特异性结合时间，提高抑制剂的选择性。首先，申请人利用伞形采样解析不同抑制剂的解离路径，结合高斯加速分子动力学模拟和马尔科夫状态模型阐明c-Src抑制剂产生不同解离速率的结构基础。其次，基于模拟结果指导化合物结构改造，结合动力学实验比较化合物与c-Src及其同亚家族其它激酶的动力学性质，优化c-Src抑制剂的动力学选择性。最后，将与c-Src不同结合时长的化合物通过细胞、器官和动物三个层面评价其抑制活性及相关副作用。

Autosomal dominant polycystic kidney disease (ADPKD) is characterized by high incidence, long disease course and lack of effective drugs. The progression of ADPKD is closely related to the activity of the c-Src kinase, which therefore represents an important target for the treatment of ADPKD. Existing c-Src inhibitors have serious side effects due to their low selectivity. Thus, it is desired to develop novel c-Src inhibitors with better selectivity. However, given its inherent similarity in the binding domain of the c-Src subfamily, it is difficult to design highly selective c-Src inhibitors based on the traditional thermodynamic parameter, i.e., affinity. In this project the applicant focuses on the dynamic dissociation process of c-Src inhibitors, aiming to optimize the duration of compounds binding to c-Src to improve the selectivity of c-Src inhibitors. Firstly, the applicant plans to use umbrella sampling to sample the dissociation pathway of different inhibitors, in combination with Gaussian accelerated molecular dynamics simulations and Markov state models, to elucidate the structural basis of ligand dissociating from the c-Src kinase. Secondly, structural modifications will be carried out based on the result of molecular dynamics simulations. Kinetic profiling of the newly synthesized agents will be further carried out at both c-Src and other kinases in the subfamily to optimize their kinetic selectivity. Finally, compounds with different receptor residence time will be evaluated for their inhibitory activity and related side effects in both in vitro and in vivo models.