Gonzaga Daniel Tadeu Gomes, Ferreira Leonardo Braga Gomes, Moreira Maramaldo Costa Thadeu Estevam, von Ranke Natalia Lidmar, Anastácio Furtado Pacheco Paulo, Sposito Simões Ana Paula, Arruda Juliana Carvalho, Dantas Luiza Pereira, de Freitas Hércules Rezende, de Melo Reis Ricardo Augusto, Penido Carmen, Bello Murilo Lamim, Castro Helena Carla, Rodrigues Carlos Rangel, Ferreira Vitor Francisco, Faria Robson Xavier, da Silva Fernando de Carvalho
Fundação Oswaldo Cruz, Instituto de Tecnologia em Fármacos, Farmanguinhos - Fiocruz, Departamento de Síntese de Fármacos Manguinhos, CEP 21041-250, Rio de Janeiro, RJ, Brazil; Universidade Federal Fluminense, Departamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, CEP 24020-150, Niterói, RJ, Brazil.
Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Laboratório de Inflamação, Avenida Brasil 4365, Manguinhos, CEP 21045-900, Rio de Janeiro, RJ, Brazil.
Eur J Med Chem. 2017 Oct 20;139:698-717. doi: 10.1016/j.ejmech.2017.08.034. Epub 2017 Aug 16.
Fifty-one 1,2,3-triazole derivatives were synthesized and evaluated with respect to P2X7 receptor (P2X7R) activity and its associated pore. These triazoles were screened in vitro for dye uptake assay and its cytotoxicity against mammalian cell types. Seven 1,2,3-triazole derivatives (5e, 6e, 8h, 9d, 9i, 11, and 12) potently blocked P2X7 receptor pore formation in vitro (J774.G8 cells and peritoneal macrophages). All blockers displayed IC value inferior to 500 nM, and they have low toxicity in either cell types. These seven selected triazoles inhibited P2X7R mediated interleukin-1 (IL-1β) release. In particular, compound 9d was the most potent P2X7R blocker. Additionally, in mouse acute models of inflammatory responses induced by ATP or carrageenan administration in the paw, compound 9d promoted a potent blocking response. Similarly, 9d also reduced mouse LPS-induced pleurisy cellularity. In silico predictions indicate this molecule appropriate to develop an anti-inflammatory agent when it was compared to commercial analogs. Electrophysiological studies suggest a competitive mechanism of action of 9d to block P2X7 receptor. Molecular docking was performed on the ATP binding site in order to observe the preferential interaction pose, indicating that binding mode of the 9d is by interacting its 1,2,3-triazole and ether moiety with positively charged residues and with its chlorobenzene moiety orientated toward the apolar end of the ATP binding site which are mainly composed by the Ile170, Trp167 and Leu309 residues from α subunit. These results highlight 9d derivative as a drug candidate with potential therapeutic application based on P2X7 receptor blockade.
合成了51种1,2,3 - 三唑衍生物，并针对P2X7受体（P2X7R）活性及其相关孔道进行了评估。这些三唑类化合物在体外进行了染料摄取试验及其对哺乳动物细胞类型的细胞毒性筛选。七种1,2,3 - 三唑衍生物（5e、6e、8h、9d、9i、11和12）在体外（J774.G8细胞和腹腔巨噬细胞）有效阻断P2X7受体孔道形成。所有阻断剂的IC值均低于500 nM，且在两种细胞类型中均具有低毒性。这七种选定的三唑类化合物抑制P2X7R介导的白细胞介素 - 1（IL - 1β）释放。特别是，化合物9d是最有效的P2X7R阻断剂。此外，在小鼠爪部注射ATP或角叉菜胶诱导的急性炎症反应模型中，化合物9d表现出强烈的阻断反应。同样，9d也降低了小鼠LPS诱导的胸膜炎细胞数量。计算机模拟预测表明，与市售类似物相比，该分子适合开发一种抗炎剂。电生理研究表明9d阻断P2X7受体的作用机制为竞争性。对ATP结合位点进行了分子对接，以观察优先相互作用姿态，表明9d的结合模式是通过其1,2,3 - 三唑和醚部分与带正电荷的残基相互作用，其氯苯部分朝向ATP结合位点的非极性末端，该位点主要由α亚基的Ile170、Trp167和Leu309残基组成。这些结果突出了9d衍生物作为基于P2X7受体阻断具有潜在治疗应用的候选药物。
