Discovery and structural optimization of 1-phenyl-3-(1-phenylethyl)urea derivatives as novel inhibitors of CRAC channel.
Discovery and structural optimization of 1-phenyl-3-(1-phenylethyl)urea derivatives as novel inhibitors of CRAC channel.
复制标题
作为新型 CRAC 通道抑制剂的 1-苯基-3-(1-苯乙基)脲衍生物的发现和结构优化。
DOI：
10.1038/aps.2015.52
复制
发表时间：
2015
影响因子：
8.2
通讯作者：
Nan Fa-jun
中科院分区：
文献类型：
--
作者：
Zhang Hai-zhen;Xu Xiao-lan;Chen Hua-yan;Ali Sher;Wang Dan;Yu Jun-wei;Xu Tao;Nan Fa-jun
目的：ca2 +释放激活ca2 +(CRAC)通道是由钙释放激活钙调节剂1 （ORAI1）形成，并由基质相互作用分子1 （STIM1）控制的存储操作通道亚家族。CRAC通道可能成为治疗免疫疾病和过敏的新靶点。本研究的目的是鉴定新的小分子CRAC通道抑制剂。方法：采用稳定共表达ORAI1和STIM1的HEK293细胞进行高通量筛选。1-苯基-3-（1-苯乙基）尿素通过ORAI1抑制CRAC通道。设计合成了其5个系列衍生物，并分析了它们的一级构效关系。我们评估了所有衍生物对Ca 2+通过CRAC通道内流HEK293细胞的影响、Jurkat细胞的细胞毒性以及表达ORAI1-SS-eGFP的Jurkat细胞中IL-2的产生。结果：通过高通量筛选，在包含32000个化合物的文库中共发现了19个hit。1-苯基-3-（1-苯乙基）尿素抑制ca2 +内流，ic50为3.25±0.17 μmol/L。对其衍生物的SAR研究表明，左侧苯胺(R1) α-位置上的烷基取代基是抑制ca2 +内流所必需的，且s构型优于r构型。右边R3被供电子基取代的衍生物比那些被吸电子基取代的衍生物表现出更强的抑制活性。此外，尿素的游离N-H并不是维持ca2 +内流抑制效能所必需的。N, N ' -二取代或N ' -取代衍生物显示出相对较低的细胞毒性，但保持了抑制IL-2产生的能力。其中化合物5b对IL-2的抑制作用较强，细胞毒性较低。结论：1-苯基-3-（1-苯乙基）尿素是一种特异性靶向ORAI1的新型CRAC通道抑制剂。本研究为设计和开发具有更好的ca2 +内流抑制、免疫抑制和低细胞毒性的CRAC通道抑制剂提供了新的化学支架。
Aim:Ca 2+-release-activated Ca 2+(CRAC) channel, a subfamily of store-operated channels, is formed by calcium release-activated calcium modulator 1 (ORAI1), and gated by stromal interaction molecule 1 (STIM1). CRAC channel may be a novel target for the treatment of immune disorders and allergy. The aim of this study was to identify novel small molecule CRAC channel inhibitors.Methods:HEK293 cells stably co-expressing both ORAI1 and STIM1 were used for high-throughput screening. A hit, 1-phenyl-3-(1-phenylethyl) urea, was identified that inhibited CRAC channels by targeting ORAI1. Five series of its derivatives were designed and synthesized, and their primary structure-activity relationships (SARs) were analyzed. All derivatives were assessed for their effects on Ca 2+ influx through CRAC channels on HEK293 cells, cytotoxicity in Jurkat cells, and IL-2 production in Jurkat cells expressing ORAI1-SS-eGFP.Results:A total of 19 hits were discovered in libraries containing 32 000 compounds using the high-throughput screening. 1-Phenyl-3-(1-phenylethyl) urea inhibited Ca 2+ influx with IC 50 of 3.25±0.17 μmol/L. SAR study on its derivatives showed that the alkyl substituent on the α-position of the left-side benzylic amine (R1) was essential for Ca 2+ influx inhibition and that the S-configuration was better than the R-configuration. The derivatives in which the right-side R3 was substituted by an electron-donating group showed more potent inhibitory activity than those that were substituted by electron-withdrawing groups. Furthermore, the free N–H of urea was not necessary to maintain the high potency of Ca 2+ influx inhibition. The N, N′-disubstituted or N′-substituted derivatives showed relatively low cytotoxicity but maintained the ability to inhibit IL-2 production. Among them, compound 5b showed an improved inhibition of IL-2 production and low cytotoxicity.Conclusion:1-Phenyl-3-(1-phenylethyl) urea is a novel CRAC channel inhibitor that specifically targets ORAI1. This study provides a new chemical scaffold for design and development of CRAC channel inhibitors with improved Ca 2+ influx inhibition, immune inhibition and low cytotoxicity.