Department of Pharmacy, University of Salerno , Via G. Paolo II 132, 84084, Fisciano, Salerno, Italy.
Department of Pharmacy, University Federico II of Naples , Via D. Montesano 49, 80131, Naples, Italy.
J Med Chem. 2016 Mar 10;59(5):2179-91. doi: 10.1021/acs.jmedchem.5b01914. Epub 2016 Feb 17.
Pharmacological modulation of the transient receptor potential melastatin type 8 (TRPM8) is currently under investigation as a new approach for the treatment of pain and other diseases. In this study, a series of N-substituted tryptamines was prepared to explore the structural requirements determining TRPM8 modulation. Using a fluorescence-based screening assay, we identified two compounds acting as an activator (2-(1H-indol-3-yl)-N-(4-phenoxybenzyl)ethanamine, 21) or an inhibitor (N,N-dibenzyl-2-(1H-indol-3-yl)ethanamine, 12) of calcium influx in HEK293 cells. In patch-clamp recordings, compound 21 displayed a significantly higher potency (EC50 = 40 ± 4 μM) and a similar efficacy when compared to menthol; by contrast, compound 12 produced a concentration-dependent inhibition of menthol-induced TRPM8 currents (IC50 = 367 ± 24 nM). Molecular modeling studies using a homology model of a single rat TRPM8 subunit identified a putative binding site located between the VSD and the TRP box, disclosing differences in the binding modes for the agonist and the antagonist.
目前，对瞬时受体电位 melastatin 型 8（TRPM8）进行药理学调节，作为治疗疼痛和其他疾病的新方法正在研究中。在这项研究中，制备了一系列 N-取代色胺，以探索决定 TRPM8 调节的结构要求。使用基于荧光的筛选测定法，我们鉴定了两种在 HEK293 细胞中作为钙内流的激活剂（2-（1H-吲哚-3-基）-N-（4-苯氧基苄基）乙胺，21）或抑制剂（N，N-二苄基-2-（1H-吲哚-3-基）乙胺，12）的化合物。在膜片钳记录中，与薄荷醇相比，化合物 21 显示出更高的效力（EC50 = 40 ± 4 μM）和相似的效力；相比之下，化合物 12 产生了薄荷醇诱导的 TRPM8 电流的浓度依赖性抑制（IC50 = 367 ± 24 nM）。使用大鼠单个 TRPM8 亚基的同源模型进行的分子建模研究确定了位于 VSD 和 TRP 盒之间的假定结合位点，揭示了激动剂和拮抗剂的结合模式的差异。
