疼痛是最常见的身体症状,也是寻求医疗护理的主要原因。长期影响人们的心理健康和社会生活。CaV3.2通道在疼痛状态的外周处理维持中起着至关重要的作用。本研究旨在鉴定靶向CaV3.2通道的新型候选药物。全细胞膜片钳,细胞热转移测定,FlexStation,体内和体外CaV3.2敲低,定点诱变,和双突变周期分析用于探索疼痛相关受体和配体-受体直接相互作用。我们发现toddaculin可以有效抑制CaV3.2通道,并显着降低背根神经节神经元的兴奋性和疼痛行为。香豆素的羰基通过范德华力(VDW)直接与CaV3.2的孔域相互作用。与结合袋的对接进一步引导我们识别甘氨酰香豆素,与母体化合物相比,对CaV3.2通道的抑制作用更强,镇痛活性更好。Toddaculin及其类似物在疼痛模型中显示出有益的治疗效果。CaV3.2上的Toddaculin结合袋可能是药物设计的有希望的对接位点。
Pain is the most common presenting physical symptom and a primary reason for seeking medical care, which chronically affects people\'s mental health and social life. CaV3.2 channel plays an essential role in the peripheral processing maintenance of pain states. This study was designed to identify novel drug candidates targeting the CaV3.2 channel. Whole-cell patch-clamp, cellular thermal shift assay, FlexStation, in vivo and in vitro CaV3.2 knock-down, site-directed mutagenesis, and double-mutant cycle analysis were employed to explore the pain-related receptors and ligand-receptor direct interaction. We found that toddaculin efficiently inhibits the CaV3.2 channel and significantly reduced the excitability of dorsal root ganglion neurons and pain behaviors. The Carbonyl group of coumarins directly interacts with the pore domain of CaV3.2 via van der Waals (VDW) force. Docking with binding pockets further led us to identify glycycoumarin, which exhibited more potent inhibition on the CaV3.2 channel and better analgesic activity than the parent compound. Toddaculin and its analog showed beneficial therapeutic effects in pain models. Toddaculin binding pocket on CaV3.2 might be a promising docking site for the design of drugs.