State Key Laboratory of Fine Chemicals, Department of Pharmaceutical Engineering, School of Chemical Engineering, Dalian University of Technology, 2 Linggong Road, Dalian 116024, PR China.
State Key Laboratory of Fine Chemicals, Department of Pharmaceutical Engineering, School of Chemical Engineering, Dalian University of Technology, 2 Linggong Road, Dalian 116024, PR China.
Bioorg Chem. 2024 Jul;148:107453. doi: 10.1016/j.bioorg.2024.107453. Epub 2024 May 14.
Thirty-five trifluoromethyl hydrazones and seventeen trifluoromethyl oxime esters were designed and synthesized via molecular hybridization. All the target compounds were initially screened for in vitro anti-inflammatory activity by assessing their inhibitory effect on NO release in LPS-stimulated RAW264.7 cells, and the optimal compound was finally identified as 2-(3-Methoxyphenyl)-N'-((6Z,9Z,12Z,15Z)-1,1,1-trifluorohenicosa-6,9,12,15-tetraen-2-ylidene)acetohydrazide (F26, IC = 4.55 ± 0.92 μM) with no cytotoxicity. Moreover, F26 potently reduced the production of PGE in LPS-stimulated RAW264.7 cells compared to indomethacin. The interaction of F26 with COX-2 and cPLA was directly verified by the CETSA technique. F26 was found to modulate the phosphorylation levels of p38 MAPK and NF-κB p65, as well as the protein expression of IκB, cPLA, COX-2, and iNOS in LPS-stimulated rat peritoneal macrophages. Additionally, F26 was observed to prevent the nuclear translocation of NF-κB p65 in LPS-stimulated rat peritoneal macrophages by immunofluorescence localization. Therefore, the aforementioned in vitro experiments demonstrated that F26 blocked the p38 MAPK and NF-κB pathways by binding to COX-2 and cPLA. In the adjuvant-induced arthritis model, F26 demonstrated a significant effect in preventing arthritis symptoms and inflammatory status in rats, exerting an immunomodulatory role by regulating the homeostasis between Th17 and Treg through inhibition of the p38 MAPK/cPLA/COX-2/PGE and NF-κB pathways. Encouragingly, F26 caused less acute ulcerogenicity in rats at a dose of 50 mg/kg compared to indomethacin. Overall, F26 is a promising candidate worthy of further investigation for treating inflammation and associated pain with lesser gastrointestinal irritation, as well as other symptoms in which cPLA and COX-2 are implicated in the pathophysiology.
设计并合成了 35 个三氟甲基腙和 17 个三氟甲基肟酯，通过分子杂交。所有目标化合物最初通过评估它们对 LPS 刺激的 RAW264.7 细胞中 NO 释放的抑制作用来筛选体外抗炎活性，最终确定最佳化合物为 2-(3-甲氧基苯基)-N'-((6Z,9Z,12Z,15Z)-1,1,1-三氟十六碳-6,9,12,15-四烯-2-亚基)乙二酰肼（F26，IC=4.55±0.92μM），无细胞毒性。此外，与吲哚美辛相比，F26 能有效降低 LPS 刺激的 RAW264.7 细胞中 PGE 的产生。CETSA 技术直接验证了 F26 与 COX-2 和 cPLA 的相互作用。F26 被发现调节 LPS 刺激的大鼠腹腔巨噬细胞中 p38 MAPK 和 NF-κB p65 的磷酸化水平，以及 IκB、cPLA、COX-2 和 iNOS 的蛋白表达。此外，通过免疫荧光定位观察到 F26 可防止 LPS 刺激的大鼠腹腔巨噬细胞核内 NF-κB p65 的易位。因此，上述体外实验表明，F26 通过与 COX-2 和 cPLA 结合阻断 p38 MAPK 和 NF-κB 途径。在佐剂诱导的关节炎模型中，F26 显著抑制关节炎症状和大鼠炎症状态，通过抑制 p38 MAPK/cPLA/COX-2/PGE 和 NF-κB 途径调节 Th17 和 Treg 之间的平衡，发挥免疫调节作用。令人鼓舞的是，与吲哚美辛相比，F26 在 50mg/kg 剂量下在大鼠中引起的急性溃疡形成性较小。总的来说，F26 是一种很有前途的候选药物，值得进一步研究，以治疗炎症及其相关疼痛，同时减少胃肠道刺激以及其他涉及 cPLA 和 COX-2 的病理生理学症状。
