Identification of endogenous surrogate ligands for human P2Y receptors through an in silico search
Identification of endogenous surrogate ligands for human P2Y receptors through an in silico search
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DOI：
10.1254/jphs.95.81
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发表时间：
2004-05-01
影响因子：
3.5
通讯作者：
Fujita, N
中科院分区：
文献类型：
--
作者：
Hiramoto, T;Nonaka, Y;Fujita, N
G蛋白偶联受体（GPCR）广泛分布于人体各处，目前近50%的药物作用于GPCR。GPCR被认为由七个跨膜α-螺旋组成，其形成α-螺旋束，其中激动剂和拮抗剂结合。靶GPCR的3D结构对于设计作用于GPCR的新药是必不可少的。我们之前已经构建了人P2 Y（1）（hP 2 Y（1））受体的3D结构，GPCR，通过同源建模与牛视紫红质的3D结构作为模板。在本研究中，我们使用AutoDock 3.0对可以结合hP 2 Y（1）受体模型的化合物进行了计算机模拟筛选。我们从30种排名靠前的化合物中选择了21种，通过测定细胞内Ca 2+浓度，我们鉴定了12种激活或阻断重组CHO细胞中稳定表达的hP 2 Y（1）受体的化合物。发现5-磷酸核糖基-1-焦磷酸（PPPP）以15 nM的低ED 50值激活hP 2 Y（1）受体。Ca ~（2+）测定表明，它对P_2Y（2）、P_2Y（6）或P_2X（2）受体没有显著影响，但对P_2Y（12）受体起弱激动剂的作用。这是第一项合理鉴定P2 Y受体家族替代配体的研究。
G protein-coupled receptors (GPCRs) are distributed widely throughout the human body, and nearly 50% of current medicines act on a GPCR. GPCRs are considered to consist of seven transmembrane a-helices that form an a-helical bundle in which agonists and antagonists bind. A 3D structure of the target GPCR is indispensable for designing novel medicines acting on a GPCR. We have previously constructed the 3D structure of human P2Y(1) (hP2Y(1)) receptor, a GPCR, by homology modeling with the 3D structure of bovine rhodopsin as a template. In the present study, we have employed an in silico screening for compounds that could bind to the hP2Y(1)-receptor model using AutoDock 3.0. We selected 21 of the 30 top-ranked compounds, and by measuring intracellular Ca2+ concentration, we identified 12 compounds that activated or blocked the hP2Y(1) receptor stably expressed in recombinant CHO cells. 5-Phosphoribosyl-1-pyrophosphate (PPPP) was found to activate the hP2Y(1) receptor with a low ED50 value of 15 nM. The Ca2+ assays showed it had no significant effect on P2Y(2), P2Y(6), or P2X(2) receptors, but acted as a weak agonist on the P2Y(12) receptor. This is the first study to rationally identify surrogate ligands for the P2Y-receptor family.