PDF(Pubmed)
Abstract：
TRP channels are important pharmacological targets in physiopathology. TRPV2 plays distinct roles in cardiac and neuromuscular function, immunity, and metabolism, and is associated with pathologies like muscular dystrophy and cancer. However, TRPV2 pharmacology is unspecific and scarce at best. Using in silico similarity-based chemoinformatics we obtained a set of 270 potential hits for TRPV2 categorized into families based on chemical nature and similarity. Docking the compounds on available rat TRPV2 structures allowed the clustering of drug families in specific ligand binding sites. Starting from a probenecid docking pose in the piperlongumine binding site and using a Gaussian accelerated molecular dynamics approach we have assigned a putative probenecid binding site. In parallel, we measured the EC50 of 7 probenecid derivatives on TRPV2 expressed in Pichia pastoris using a novel medium-throughput Ca2+ influx assay in yeast membranes together with an unbiased and unsupervised data analysis method. We found that 4-(piperidine-1-sulfonyl)-benzoic acid had a better EC50 than probenecid, which is one of the most specific TRPV2 agonists to date. Exploring the TRPV2-dependent anti-hypertensive potential in vivo, we found that 4-(piperidine-1-sulfonyl)-benzoic acid shows a sex-biased vasodilator effect producing larger vascular relaxations in female mice. Overall, this study expands the pharmacological toolbox for TRPV2, a widely expressed membrane protein and orphan drug target.
摘要：
TRP通道是病理生理学中的重要药理靶点。TRPV2在心脏和神经肌肉功能中起着不同的作用，豁免权,和新陈代谢，并与肌肉营养不良和癌症等病理有关。然而,TRPV2药理学是无特异性和稀缺的。使用基于计算机相似性的化学信息学，我们获得了基于化学性质和相似性分为家族的TRPV2的270个潜在命中。将化合物对接在可用的大鼠TRPV2结构上允许药物家族聚集在特定的配体结合位点中。从胡椒长胺结合位点的丙磺舒对接姿势开始，并使用高斯加速分子动力学方法，我们分配了一个推定的丙磺舒结合位点。并行,我们使用一种新型的酵母膜中中等通量Ca2内流测定法以及无偏和无监督的数据分析方法，测量了7种丙磺舒衍生物在毕赤酵母中表达的TRPV2上的EC50。我们发现4-(哌啶-1-磺酰基)-苯甲酸比丙磺舒具有更好的EC50,这是迄今为止最具体的TRPV2激动剂之一。探索体内TRPV2依赖性抗高血压潜能，我们发现4-(哌啶-1-磺酰基)-苯甲酸显示出性别偏倚的血管舒张作用,在雌性小鼠中产生更大的血管舒张作用.总的来说,这项研究扩展了TRPV2的药理学工具箱，TRPV2是一种广泛表达的膜蛋白和孤儿药物靶标。
