title: Biological activity of a small molecule indole analog, 1-[(1H-indol-3-yl)methylene]-2-phenylhydrazine (HMPH), in chronic inflammation.
abstract: 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.
authors.affiliation: Department of Pharmacy Practice, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Chandra Sekhar
authors.initials: CS
authors.lastname: Misra
authors.affiliation: Department of Pharmacy Practice, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Chethan
authors.initials: C
authors.lastname: Gejjalagere Honnappa
authors.affiliation: Department of Pharmacognosy, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Srinivas Reddy
authors.initials: SR
authors.lastname: Jitta
authors.affiliation: Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Karthik
authors.initials: K
authors.lastname: Gourishetti
authors.affiliation: Department of Pharmacognosy, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Prasanthi
authors.initials: P
authors.lastname: Daram
authors.affiliation: Department of Pharmacology, Life Science Research Centre in India, Daiichi Sankyo India Pharma Private Limited, Gurgaon, 122015, India.
authors.forename: Mahendra Pal
authors.initials: MP
authors.lastname: Singh
authors.affiliation: Department of Pharmacy Practice, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Akhila
authors.initials: A
authors.lastname: Hosur Shrungeswara
authors.affiliation: Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India.
authors.forename: Yogendra
authors.initials: Y
authors.lastname: Nayak
authors.affiliation: Department of Pharmacy Practice, Manipal College of Pharmaceutical Sciences, Manipal University, Karnataka, Manipal, 576104, India. Electronic address: mkunnikrishnan@gmail.com.
authors.forename: Mazhuvancherry Kesavan
authors.initials: MK
authors.lastname: Unnikrishnan
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D002351:Carrageenan; D002478:Cells, Cultured; D002908:Chronic Disease; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D006835:Hydrazones / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D007211:Indoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D007249:Inflammation / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D015227:Lipid Peroxidation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects; D008297:Male; D051379:Mice; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000096:biosynthesis
chemical_list: C000608564:1-((1H-indol-3-yl)methylene)-2-phenylhydrazine; D000894:Anti-Inflammatory Agents, Non-Steroidal; D006835:Hydrazones; D007211:Indoles; D017382:Reactive Oxygen Species; D054852:Small Molecule Libraries; D014409:Tumor Necrosis Factor-alpha; D002351:Carrageenan
keywords: Anti-inflammatory; Arthritis; Carrageenan; HMPH [1-((1H-indol-3-yl) methylene)-2-phenylhydrazine]; Lipopolysaccharide; TNF-α