Misra Chandra Sekhar, Gejjalagere Honnappa Chethan, Jitta Srinivas Reddy, Gourishetti Karthik, Daram Prasanthi, Singh Mahendra Pal, Hosur Shrungeswara Akhila, Nayak Yogendra, Unnikrishnan Mazhuvancherry Kesavan
Department of Pharmacy Practice, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
Department of Pharmacognosy, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
Chem Biol Interact. 2016 Jan 25;244:71-83. doi: 10.1016/j.cbi.2015.10.024. Epub 2015 Nov 5.
A synthetic small molecule, 1-[(1H-indol-3-yl)methylene]-2-phenylhydrazine (HMPH) was conveniently synthesised by a one-step reaction, purified and characterised by chromatographic and spectroscopic methods. HMPH scavenged free radicals and inhibited lipopolysaccharide (LPS)-induced ROS generation and NO release in RAW-264.7 cells without signs of any detectable cytotoxicity. HMPH inhibited lipid peroxidation (LPO) with IC50 of 135 ± 9 as against 58 ± 8 μM for α-tocopherol. Further, HMPH (>50 μM) significantly reduced the LPS-induced TNF-α release in mouse peritoneal macrophages and in human peripheral blood mononuclear cells (PBMCs). HMPH did not show any visible signs of toxicity in rats up to 400 mg/kg/intraperitoneal and 2000 mg/kg/oral. HMPH at 25 and 50 mg/kg attenuated neutrophil infiltration in air-pouch lavage and bronchoalveolar lavage (BAL) in rat models. HMPH also reduced myeloperoxidase (MPO), nitrite and TNF-α in air-pouch lavage in addition to MPO in plasma. HMPH reduced acute paw-inflammation in carrageenan-induced paw-edema. HMPH consistently decreased both ipsilateral and contralateral paw inflammation, minimised the clinical scores of arthritis, prevented body weight (B.wt.) loss, attenuated serum C-reactive protein (C-RP) and rheumatoid factors (RF) in rat model of adjuvant-induced arthritis. Histopathology and radio-graphical reports show that HMPH reduced bone erosion in both ipsilateral and contralateral paw joints. Failure to inhibit COX suggests that effectiveness of HMPH in both acute and chronic inflammation is mediated by a multimodal mechanism involving modulation of immunity, attenuating TNF-α, protecting bone attrition and reducing oxidative stress.
一种合成小分子1-[(1H-吲哚-3-基)亚甲基]-2-苯基肼（HMPH）通过一步反应简便合成，经色谱和光谱方法纯化与表征。HMPH可清除自由基，抑制脂多糖（LPS）诱导的RAW-264.7细胞中活性氧（ROS）生成及一氧化氮（NO）释放，且无任何可检测到的细胞毒性迹象。HMPH抑制脂质过氧化（LPO）的IC50为135±9 μM，而α-生育酚为58±8 μM。此外，HMPH（>50 μM）可显著降低LPS诱导的小鼠腹腔巨噬细胞及人外周血单核细胞（PBMCs）中肿瘤坏死因子-α（TNF-α）的释放。在大鼠中，腹腔注射400 mg/kg及口服2000 mg/kg剂量的HMPH均未显示出任何明显的毒性迹象。在大鼠模型中，25和50 mg/kg剂量的HMPH可减轻气袋灌洗和支气管肺泡灌洗（BAL）中的中性粒细胞浸润。HMPH还可降低气袋灌洗中的髓过氧化物酶（MPO）、亚硝酸盐和TNF-α，以及血浆中的MPO。HMPH可减轻角叉菜胶诱导的爪肿胀中的急性爪部炎症。在佐剂诱导的关节炎大鼠模型中，HMPH持续降低同侧和对侧爪部炎症，使关节炎临床评分降至最低，防止体重减轻，减轻血清C反应蛋白（C-RP）和类风湿因子（RF）。组织病理学和影像学报告显示，HMPH可减轻同侧和对侧爪关节的骨侵蚀。未能抑制环氧化酶（COX）表明，HMPH在急性和慢性炎症中的有效性是通过一种多模式机制介导的，该机制涉及免疫调节、减轻TNF-α、保护骨质流失和降低氧化应激。
