title: Novel synthetic biscoumarins target tumor necrosis factor-α in hepatocellular carcinoma in vitro and in vivo.
abstract: TNF is a pleotropic cytokine known to be involved in the progression of several pro-inflammatory disorders. Many therapeutic agents have been designed to counteract the effect of TNF in rheumatoid arthritis as well as a number of cancers. In the present study we have synthesized and evaluated the anti-cancer activity of novel biscoumarins in vitro and in vivo. Among new compounds, BIHC was found to be the most cytotoxic agent against the HepG2 cell line while exhibiting less toxicity toward normal hepatocytes. Furthermore, BIHC inhibited the proliferation of various hepatocellular carcinoma (HCC) cells in a dose- and time-dependent manner. Subsequently, using in silico target prediction, BIHC was predicted as a TNF blocker. Experimental validation was able to confirm this hypothesis, where BIHC could significantly inhibit the recombinant mouse TNF-α binding to its antibody with an IC50 of 16.5 μM. Furthermore, in silico docking suggested a binding mode of BIHC similar to a ligand known to disrupt the native, trimeric structure of TNF, and also validated with molecular dynamics simulations. Moreover, we have demonstrated the down-regulation of p65 phosphorylation and other NF-κB-regulated gene products upon BIHC treatment, and on the phenotypic level the compound shows inhibition of CXCL12-induced invasion of HepG2 cells. Also, we demonstrate that BIHC inhibits infiltration of macrophages to the peritoneal cavity and suppresses the activity of TNF-α in vivo in mice primed with thioglycollate broth and lipopolysaccharide. We comprehensively validated the TNF-α inhibitory efficacy of BIHC in an inflammatory bowel disease mice model.
authors.affiliation: Laboratory of Chemical Biology, Department of Chemistry, Bangalore University, Palace Road, Bangalore 560 001, India.
authors.forename: Hosadurga Kumar
authors.initials: HK
authors.lastname: Keerthy
authors.affiliation: Department of Studies in Chemistry and University of Mysore, Manasagangotri, Mysore 570 006, India.
authors.forename: Chakrabhavi Dhananjaya
authors.initials: CD
authors.lastname: Mohan
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117 597.
authors.forename: Kodappully Sivaraman
authors.initials: KS
authors.lastname: Siveen
authors.affiliation: Unilever Centre for Molecular Science Informatics, Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Julian E
authors.initials: JE
authors.lastname: Fuchs
authors.affiliation: Interdisciplinary Research Group of Infectious Diseases, Singapore-MIT Alliance for Research and Technology Centre (SMART), Singapore 138 602, and.
authors.forename: Shobith
authors.initials: S
authors.lastname: Rangappa
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysore 570 006, India.
authors.forename: Mahalingam S
authors.initials: MS
authors.lastname: Sundaram
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117 597.
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysore 570 006, India.
authors.forename: Kesturu S
authors.initials: KS
authors.lastname: Girish
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117 597,; Cancer Science Institute of Singapore, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117 599.
authors.forename: Gautam
authors.initials: G
authors.lastname: Sethi
authors.affiliation: Laboratory of Chemical Biology, Department of Chemistry, Bangalore University, Palace Road, Bangalore 560 001, India,. Electronic address: salundibasappa@gmail.com.
authors.lastname: Basappa
authors.affiliation: Unilever Centre for Molecular Science Informatics, Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom,. Electronic address: ab454@cam.ac.uk.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bender
authors.affiliation: Department of Studies in Chemistry and University of Mysore, Manasagangotri, Mysore 570 006, India. Electronic address: rangappaks@yahoo.com.
authors.forename: Kanchugarakoppal Subbegowda
authors.initials: KS
authors.lastname: Rangappa
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000378:metabolism; D003374:Coumarins / Q000737:chemistry*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D015195:Drug Design; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects; D006801:Humans; D007249:Inflammation; D015212:Inflammatory Bowel Diseases / Q000378:metabolism; D020128:Inhibitory Concentration 50; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D062105:Molecular Docking Simulation; D016328:NF-kappa B / Q000378:metabolism; D011485:Protein Binding; D015398:Signal Transduction; D020349:Surface Plasmon Resonance; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry*
chemical_list: D000893:Anti-Inflammatory Agents; D003374:Coumarins; D016207:Cytokines; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha
keywords: Cancer Therapy; Cell Invasion; Hepatocellular Carcinoma; Inflammatory Bowel Disease (IBD); NF-kappa B (NF-KB); Tumor Necrosis Factor (TNF)