Metastin (KiSS-1) mimetics identified from peptide structure-activity relationship-derived pharmacophores and directed small molecule database screening
Metastin (KiSS-1) mimetics identified from peptide structure-activity relationship-derived pharmacophores and directed small molecule database screening
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DOI：
10.1021/jm0609824
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发表时间：
2007-02-08
影响因子：
7.3
通讯作者：
Mayo, Kevin H.
中科院分区：
文献类型：
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作者：
Orsini, Michael J.;Klein, Mark A.;Mayo, Kevin H.
转移蛋白，也称为KiSS-1，是转移蛋白受体GPR 54的同源配体，是已知在实验模型中显著减少转移的肽。尽管如此，没有报道的转移蛋白的结构，也没有任何可以在临床或实验上使用的转移蛋白功能的小分子调节剂。在这里，我们报告的NMR溶液结构的13个残基的metastin肽在膜样环境（SDS胶束），并发现它有一个相对稳定的螺旋构象从残基7至13。在metastin受体结合和钙流与受体转染的HEK-293细胞的测定中，我们通过丙氨酸扫描和氨基酸取代证明肽C-末端在NMR结构模型中显示螺旋周期性，并且Phe 9，Arg 12和Phe 13对肽的活性至关重要。这三个残基位于螺旋的一面，并定义了metastin的药效团位点。我们在小分子数据库搜索中使用这些药效团特征来鉴定对metastin受体具有亚微摩尔亲和力的命中。我们还表明，模拟该药效团位点的关键元素的分子与metastin受体结合，并作为完全激动剂，尽管与metastin本身相比效力降低。总之，这种结构-活性方法可以产生与定义和调节转移素受体功能相关的非常有用的化合物。
Metastin, also known as KiSS-1, the cognate ligand for the metastin receptor GPR54, is a peptide known to dramatically reduce metastasis in experimental models. Despite this, there is no reported structure for metastin nor any small molecule modulators of metastin function that could be used either clinically or experimentally. Here we report the NMR solution structure of a 13-residue metastin peptide in a membrane-like environment (SDS micelles) and find it to have a relatively stable helix conformation from residues 7 to 13. In assays for metastin receptor binding and calcium flux with receptor-transfected HEK-293 cells, we demonstrate through alanine scanning and amino acid substitutions that the peptide C-terminus shows helix periodicity in an NMR structural model and that Phe9, Arg12, and Phe13 are crucial to the activity of the peptide. These three residues lie on one face of the helix and define a pharmacophore site for metastin. We used these pharmacophore features in small molecule database searches to identify hits with submicromolar affinity for the metastin receptor. We also show here that molecules mimicking key elements of this pharmacophore site bind to the metastin receptor and act as full agonists, albeit with reduced potency compared to that of metastin itself. Together this structure-activity approach may yield pharmacologically useful compounds relevant in defining and modulating metastin receptor function.