Department of Pharmaceutical Chemistry, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy.
J Med Chem. 2010 May 13;53(9):3489-501. doi: 10.1021/jm901691y.
The application of frontal affinity chromatography-mass spectrometry (FAC-MS), along with molecular modeling studies, to the screening of potential drug candidates toward the recently deorphanized G-protein-coupled receptor (GPCR) GPR17 is shown. GPR17 is dually activated by uracil nucleotides and cysteinyl-leukotrienes, and is expressed in organs typically undergoing ischemic damage (i.e., brain, heart and kidney), thus representing a new pharmacological target for acute and chronic neurodegeneration. GPR17 was entrapped on an immobilized artificial membrane (IAM), and this stationary phase was used to screen a library of nucleotide derivatives by FAC-MS to select high affinity ligands. The chromatographic results have been validated with a reference functional assay ([(35)S]GTPgammaS binding assay). The receptor nucleotide-binding site was studied by setting up a column where a mutated GPR17 receptor (Arg255Ile) has been immobilized. The chromatographic behavior of the tested nucleotide derivatives together with in silico studies have been used to gain insights into the structure requirement of GPR17 ligands.
本文展示了正面亲和色谱-质谱（FAC-MS）联用分子建模研究在筛选针对最近去孤儿化的 G 蛋白偶联受体（GPCR）GPR17 的潜在药物候选物中的应用。GPR17 被尿嘧啶核苷酸和半胱氨酰白三烯双重激活，在通常经历缺血损伤的器官（即脑、心脏和肾脏）中表达，因此代表了急性和慢性神经退行性变的新的药理学靶标。GPR17 被包埋在固定化人工膜（IAM）上，该固定相用于通过 FAC-MS 筛选核苷酸衍生物文库，以选择高亲和力配体。色谱结果已通过参考功能测定（[（35）S]GTPγS 结合测定）进行了验证。通过设置一个固定化突变 GPR17 受体（Arg255Ile）的柱子来研究受体核苷酸结合位点。测试的核苷酸衍生物的色谱行为以及计算机模拟研究用于深入了解 GPR17 配体的结构要求。
