Novel synthetic coumarins that targets NF-jB in Hepatocellular carcinoma Mahabaleshwaraiah Neelgundmath a,b,  , Koragere Rajashekar Dinesh c,  , Chakrabhavi Dhananjaya Mohan d,  , Feng Li e , Xiaoyun Dai e , Kodappully Sivaraman Siveen e , Shardul Paricharak f,g , Daniel J. Mason f , Julian E. Fuchs f , Gautam Sethi e , Andreas Bender f , Kanchugarakoppal S. Rangappa d , Obelannavar Kotresh b,⇑ , Basappa c,⇑ a Department of Chemistry, P.C. Jabin Science College, Hubli, India b Department of Chemistry, Karnataka Science College, Dharwad, India c Laboratory of Chemical Biology, Department of Chemistry, Bangalore University, Central College Campus, Palace Road, Bangalore 560001, India d Department of Studies in Chemistry, Manasagangotri, University of Mysore, Mysore 570005, India e Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117597 Singapore, Singapore f Centre for Molecular Science Informatics, Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW Cambridge, United Kingdom g Division of Medicinal Chemistry, Leiden Academic Centre for Drug Research, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands article info Article history: Received 31 August 2014 Revised 27 November 2014 Accepted 19 December 2014 Available online 30 December 2014 Keywords: Hepatocellular carcinoma Coumarin NF-jB Bioinformatics Inflammation abstract Hepatocellular carcinoma (HCC) is the fifth most common malignant tumor worldwide, and is the third most common cause of cancer related death. Constitutive activation of NF-jB is the underlying mecha- nism behind tumorigenesis and this protein regulates the expression of genes involved in proliferation, survival, drug resistance, angiogenesis and metastasis. The design of inhibitors which suppress NF-jB activation is therefore of great therapeutic importance in the treatment of HCC. In this study, we inves- tigated the effect of newly synthesized coumarin derivatives against HCC cells, and identified (7-Carbeth- oxyamino-2-oxo-2H-chromen-4-yl)methylpyrrolidine-1 carbodithioate (CPP) as lead compound. Further, we evaluated the effect of CPP on the DNA binding ability of NF-jB, CXCL12-induced cell migration and invasion, and the regulated gene products in HCC cells. We found that CPP induced cytotoxicity in three HCC cells in a time and dose dependent manner, and suppressed the DNA binding ability of NF-jB. CPP significantly decreased the CXCL12-induced cell migration and invasion. More evidently, CPP inhibits the expression of NF-jB targeted genes such as cyclin D1, Bcl-2, survivin, MMP12 and C-Myc. Furthermore, the molecular docking analysis suggested that CPP interacts with the p50 binding domain of the p65 sub- unit, scoring best among the 26 docked coumarin derivatives of this study. Thus, we are reporting CPP as a potent inhibitor of the pro-inflammatory pathway in Hepatocellular carcinoma. Ó 2014 Elsevier Ltd. All rights reserved. Hepatocellular carcinoma (HCC) is the most common form of liver cancer, and is the third most common cause of cancer related mortality. 1 Obesity, hepatitis B or hepatitis C virus infection, con- sumption of aflatoxin B1, and alcoholic hepatitis are the major risk factors of HCC. 2 The majority of patients with HCC are diagnosed at a late stage of the disease, with therapy limited to only liver trans- plantation and surgical liver sectioning. Early detection of HCC may contribute to the strengthening of prognosis. 3,4 Nuclear factor kappa B (NF-jB) is an inducible transcription factor present ubiqui- tously in the cytoplasm of almost all mammalian cells, and was identified by Baltimore and colleagues in 1986. 5 In an unstimulated cell, NF-jB is associated with IjB (inhibitory jB), and resides in the cytoplasm. 6 Different ligands acting upon the receptor result in a cascade of upstream kinases, finally leading to phosphorylation and ubiquitylation of IjB, which is targeted for degradation. NF- jB translocates into nucleus and regulates transcription of the genes involved in inflammation. 7 NF-jB is referred to as a dou- ble-edged sword by researchers because of its critical role in proper functioning of the immune system, and the possibility that onco- genesis may arise from the slightest change in regulation. 8 Several anti-apoptotic, angiogenic, inflammatory, metastasis-related, and http://dx.doi.org/10.1016/j.bmcl.2014.12.065 0960-894X/Ó 2014 Elsevier Ltd. All rights reserved. Abbreviation: CPP, (7-Carbethoxyamino-2-oxo-2H-chromen-4-yl)methylpyrrol- idine-1 carbodithioate. ⇑ Corresponding authors. Tel.: +91 9686449876 (Basappa). E-mail address: salundibasappa@gmail.com (Basappa).   These authors contributed equally. Bioorganic & Medicinal Chemistry Letters 25 (2015) 893–897 Contents lists available at ScienceDirect Bioorganic & Medicinal Chemistry Letters journal homepage: www.elsevier.com/locate/bmcl