Abstract：
LIM Kinases, LIMK1 and LIMK2, have become promising targets for the development of inhibitors with potential application for the treatment of several major diseases. LIMKs play crucial roles in cytoskeleton remodeling as downstream effectors of small G proteins of the Rho-GTPase family, and as major regulators of cofilin, an actin depolymerizing factor. In this article we describe the conception, synthesis, and biological evaluation of novel tetrahydropyridine pyrrolopyrimidine LIMK inhibitors. Homology models were first constructed to better understand the binding mode of our preliminary compounds and to explain differences in biological activity. A library of over 60 products was generated and in vitro enzymatic activities were measured in the mid to low nanomolar range. The most promising derivatives were then evaluated in cell on cofilin phosphorylation inhibition which led to the identification of 52 which showed excellent selectivity for LIMKs in a kinase selectivity panel. We also demonstrated that 52 affected the cell cytoskeleton by disturbing actin filaments. Cell migration studies with this derivative using three different cell lines displayed a significant effect on cell motility. Finally, the crystal structure of the kinase domain of LIMK2 complexed with 52 was solved, greatly improving our understanding of the interaction between 52 and LIMK2 active site. The reported data represent a basis for the development of more efficient LIMK inhibitors for future in vivo preclinical validation.
摘要：
LIM激酶,LIMK1和LIMK2已成为开发抑制剂的有希望的靶标，具有治疗几种主要疾病的潜在应用。LIMKs作为Rho-GTP酶家族小G蛋白的下游效应子，在细胞骨架重塑中起着至关重要的作用，作为cofilin的主要监管机构，肌动蛋白解聚因子。在这篇文章中，我们描述了这个概念，合成，新型四氢吡啶吡咯并嘧啶LIMK抑制剂的生物学评价。首先构建同源性模型以更好地理解我们的初步化合物的结合模式并解释生物活性的差异。产生超过60种产物的文库,并在中至低纳摩尔范围内测量体外酶活性。然后在细胞中评估最有前途的衍生物对cofilin磷酸化抑制的影响，从而鉴定出52种在激酶选择性组中对LIMK表现出优异的选择性。我们还证明了52通过干扰肌动蛋白丝来影响细胞骨架。使用三种不同细胞系对该衍生物进行的细胞迁移研究显示出对细胞运动性的显着影响。最后,与52复合的LIMK2激酶结构域的晶体结构得到解决，大大提高了我们对52和LIMK2活性位点之间相互作用的理解。报道的数据代表了开发更有效的LIMK抑制剂用于未来体内临床前验证的基础。
