Development of Photoactivatable Allosteric Modulators for the Chemokine Receptor CXCR3
Development of Photoactivatable Allosteric Modulators for the Chemokine Receptor CXCR3
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DOI：
10.1002/cmdc.201500573
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发表时间：
2016-03-17
期刊：
影响因子：
3.4
通讯作者：
Tschammer, Nuska
中科院分区：
文献类型：
--
作者：
Admas, Tizita Haimanot;Bernat, Viachaslau;Tschammer, Nuska
CXCR 3受体是一种A类G蛋白偶联受体（GPCR），参与各种免疫细胞的调节和运输。已经提出CXCR 3拮抗剂有益于治疗广泛的病症，包括但不限于炎性和自身免疫性疾病。然而，CXCR 3配体的基于结构的设计仍然受到缺乏详细描述变构配体与受体之间相互作用的结构信息的阻碍。我们设计并合成了用于结构和功能表征的光活化探针，使用光亲和标记，然后进行质谱分析，AMG 487和RAMX 3（两种有效的选择性CXCR 3负变构调节剂）的CXCR 3变构结合口袋。光亲和标记是一种常见的方法，通过光活化探针的帮助下，转化为极其活跃的中间体光解后，阐明GPCR的小分子配体的结合模式。光不稳定探针N-[（{1-[3-（4-乙氧基苯基）-4-氧代-3，4-二氢吡啶并[2，3-d]嘧啶-2-基]乙基}-2-[4-氟-3-（三氟甲基）苯基]-N-{1-[4-（3-（三氟甲基）-3H-二氮杂环丙烯-3-基]苄基}哌啶-4-基）甲基]乙酰胺（10）在[H-3] RAMX 3放射性配体置换测定中显示CXCR 3受体的显著标记（80%）。化合物10将用作通过质谱法详细研究CXCR 3的结合口袋的重要工具化合物。
The CXCR3 receptor, a classA Gprotein-coupled receptor (GPCR), is involved in the regulation and trafficking of various immune cells. CXCR3 antagonists have been proposed to be beneficial for the treatment of a wide range of disorders including but not limited to inflammatory and autoimmune diseases. The structure-based design of CXCR3 ligands remains, however, hampered by a lack of structural information describing in detail the interactions between an allosteric ligand and the receptor. We designed and synthesized photoactivatable probes for the structural and functional characterization, using photoaffinity labeling followed by mass spectrometry, of the CXCR3 allosteric binding pocket of AMG 487 and RAMX3, two potent and selective CXCR3 negative allosteric modulators. Photoaffinity labeling is a common approach to elucidate binding modes of small-molecule ligands of GPCRs through the aid of photoactivatable probes that convert to extremely reactive intermediates upon photolysis. The photolabile probe N-[({1-[3-(4-ethoxyphenyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-2-yl]ethyl}-2-[4-fluoro-3-(trifluoromethyl)phenyl]-N-{1-[4-(3-(trifluoromethyl)-3H-diazirin-3-yl]benzyl}piperidin-4-yl)methyl]acetamide (10) showed significant labeling of the CXCR3 receptor (80%) in a [H-3]RAMX3 radioligand displacement assay. Compound 10 will serve as an important tool compound for the detailed investigation of the binding pocket of CXCR3 by mass spectrometry.