Department of Chemical Biology and Theraputics , St. Jude Children's Research Hospital , Memphis , Tennessee 38105 United States.
Howard Hughes Medical Institute , St. Jude Children's Research Hospital , Memphis , Tennessee 38105 United States.
J Med Chem. 2018 Apr 12;61(7):2680-2693. doi: 10.1021/acs.jmedchem.7b01277. Epub 2018 Mar 26.
We previously discovered and validated a class of piperidinyl ureas that regulate defective in cullin neddylation 1 (DCN1)-dependent neddylation of cullins. Here, we report preliminary structure-activity relationship studies aimed at advancing our high-throughput screen hit into a tractable tool compound for dissecting the effects of acute DCN1-UBE2M inhibition on the NEDD8/cullin pathway. Structure-enabled optimization led to a 100-fold increase in biochemical potency and modestly increased solubility and permeability as compared to our initial hit. The optimized compounds inhibit the DCN1-UBE2M protein-protein interaction in our TR-FRET binding assay and inhibit cullin neddylation in our pulse-chase NEDD8 transfer assay. The optimized compounds bind to DCN1 and selectively reduce steady-state levels of neddylated CUL1 and CUL3 in a squamous cell carcinoma cell line. Ultimately, we anticipate that these studies will identify early lead compounds for clinical development for the treatment of lung squamous cell carcinomas and other cancers.
我们之前发现并验证了一类哌啶基脲，可调节 Cullin 连接酶 neddylation1（DCN1）依赖性 Cullin 的 neddylation 缺陷。在这里，我们报告了初步的结构-活性关系研究，旨在将高通量筛选的命中化合物推进为一种可行的工具化合物，以剖析急性 DCN1-UBE2M 抑制对 NEDD8/Cullin 途径的影响。与我们最初的命中化合物相比，结构使能优化使生化效力提高了 100 倍，并且略微提高了溶解度和通透性。优化后的化合物在我们的 TR-FRET 结合测定中抑制 DCN1-UBE2M 蛋白-蛋白相互作用，并在我们的脉冲追踪 NEDD8 转移测定中抑制 Cullin 的 neddylation。优化后的化合物与 DCN1 结合，并在鳞状细胞癌细胞系中选择性降低稳定状态下的 neddylated CUL1 和 CUL3 水平。最终，我们预计这些研究将为治疗肺鳞状细胞癌和其他癌症的临床开发确定早期先导化合物。
