分类： 生物学 >> 生物物理学 提交时间： 2016-05-11
摘要： Aim: Ca2+-release-activated Ca2+ (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 Ca2+ 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 Ca2+ influx with IC50 of 3.25 +/- 0.17 mu mol/L. SAR study on its derivatives showed that the alkyl substituent on the a-position of the left-side benzylic amine (R1) was essential for Ca2+ 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 Ca2+ 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 Ca2+ influx inhibition, immune inhibition and low cytotoxicity.
分类： 工程与技术科学 >> 生物医学工程学 提交时间： 2023-05-27
Wang, Dongping
Jin, Kai
Hu, Chenxuan
Du, Maohua
Belgaid, Yacine
Shi, Subao
Li, Jiahao
Hu, Siyi
Yu, Jun
Nathan, Arokia
Ma, Hanbin
摘要： Digital liquid sample handling is an enabling tool for cutting-edge lifesciences research. Similar to Field Programmable Gate Arrays (FPGA) in Integrated Circuits, we present here an active-matrix thin-film electronicsbased digital microfluidics system, which will be henceforth referred to as Field Programmable Droplets Arrays (FPDA). The system contains 256 256 pixels in an active area of 10.65 cm2, which is capable of manipulating thousands of individually addressable liquid droplets simultaneously. By leveraging a customised TFT-based circuit design solution, it becomes possible to programmatically manipulate droplets at the pixel level. The minimum achievable droplet volume is around 0.5 nl, which is two orders of magnitude smaller than the state-of-the-art reported1. The movement of droplets can be either pre-programmed or controlled in real-time. The FPDA system shows great potential of the ubiquitous thin-film electronics technology in digital liquid handling. These efforts will make it possible to create a true programmable lab-on-a-chip device to enable great advances in life science research.
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