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.

中文翻译：

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哌啶尿素化学控制在Cullin Neddylation 1（DCN1）介导的Cullin Neddylation中有缺陷。

我们先前发现并验证了一类哌啶基脲，它们可调节cullin扑灭1（DCN1）依赖性的neddylation中的缺陷。在这里，我们报告了初步的结构-活性关系研究，旨在将我们的高通量筛选命中率提高为易于处理的工具化合物，以剖析急性DCN1-UBE2M抑制作用对NEDD8 / cullin途径的影响。与我们最初的产品相比，结构优化使生化效能提高了100倍，溶解度和渗透性适度提高。在我们的TR-FRET结合试验中，优化的化合物可抑制DCN1-UBE2M蛋白质-蛋白质的相互作用，而在脉冲追逐NEDD8转移试验中，可抑制cullin的酰化。优化的化合物与DCN1结合，并选择性降低鳞状细胞癌细胞系中的NDM的稳态水平。最终，我们预计这些研究将确定用于治疗肺鳞状细胞癌和其他癌症的临床开发的早期先导化合物。