1 3 Anti-inflammatory effect of thalidomide dithiocarbamate and dithioate 4 analogs 5 6 7 Roba Talaat a , Waheba El-Sayed a , Hussein Agwa b , Amira Gamal-Eldeen c , Shaden Moawia a , 8 Magdy Zahran b,⇑ 9 a Department of Molecular Biology, Genetic Engineering and Biotechnology Research Institute, Sadat City University, Sadat City, Egypt 10 b Chemistry Department, Faculty of Science, Menoufiya University, Shebin El-Koam, Egypt 11 c Cancer Biology Laboratory, Center of Excellency for Advanced Sciences, National Research Centre, Dokki, Cairo, Egypt 12 13 15 article info 16 Article history: 17 Received 13 February 2015 18 Received in revised form 12 May 2015 19 Accepted 29 May 2015 20 Available online xxxx 21 Keywords: 22 Anti-inflammatory 23 Hydroxyl (OH Å ) radical 24 Thalidomide analogs 25 Prooxidant activity 26 27 abstract 28 Thalidomide has anti-inflammatory, immunomodulatory, and anti-angiogenic properties. It has been 29 used to treat a variety of cancers and autoimmune diseases. This study aimed to characterize 30 anti-inflammatory activities of novel thalidomide analogs by exploring their effects on splenocytes pro- 31 liferation and macrophage functions and their antioxidant activity. MTT assay was used to assess the 32 cytotoxic effect of thalidomide analogs against splenocytes. Tumor necrosis factor (TNF-a) and nuclear 33 factor kappa B (NF-jB-P65) were determined by enzyme-linked immunosorbent assay (ELISA). Nitric 34 oxide (NO) was estimated by colorimetric assay. Antioxidant activity was examined by ORAC assay. 35 Our results demonstrated that thalidomide dithioate analog 2 and thalidomide dithiocarbamate analog 36 4 produced a slight increase in splenocyte proliferation compared with thalidomide. Thalidomide dithio- 37 carbamate analog 1 is a potent inhibitor of TNF-a production, whereas thalidomide dithiocarbamate ana- 38 log 5 is a potent inhibitor of both TNF-a and NO. Analog 2 has a pronounced inhibitory effect on 39 NF-jB-P65 production level. All thalidomide analogs showed prooxidant activity against hydroxyl 40 (OH Å ) radical. Analog 1 and thalidomide dithioate analog 3 have prooxidant activity against peroxyl 41 (ROO Å ) radical in relation to thalidomide. On the other hand, analog 4 has a potent scavenging capacity 42 against peroxyl (ROO Å ) radical compared with thalidomide. Taken together, the results of this study sug- 43 gest that thalidomide analogs might have valuable anti-inflammatory activities with more pronounced 44 effect than thalidomide itself. 45 Ó 2015 Elsevier Ireland Ltd. All rights reserved. 46 47 48 49 1. Introduction 50 Inflammation is a physiologic process in response to tissue 51 damage resulting from microbial pathogen infection, chemical irri- 52 tation, and/or wounding [1–2]. Dysregulated inflammation plays a 53 major role in resulting chronic inflammation that can contribute to 54 diseases such as arthritis, heart attacks, Alzheimer’s disease, and 55 cancer [3]. Inflammatory responses play decisive roles at different 56 stages of tumor development [4]. Inflammation also affects 57 immune surveillance and responses to therapy [4]. Several 58 anti-inflammatory drugs have been found to reduce tumor 59 incidence when used as prophylactics, as well as to slow down 60 progression and reduce mortality when used as therapeutics [5]. 61 These drugs include nonsteroidal anti-inflammatory drugs (such 62 as cyclooxygenase (COX 2 ) inhibitors), and anti-inflammatory 63 steroids (such as dexamethasone) [6]. 64 Thalidomide has potent anti-inflammatory effects through inhi- 65 bition of tumor necrosis factor (TNF-a) and suppression of nuclear 66 factor kappa B (NF-jB) activation in response to inflammatory 67 agents [7,8]. Thalidomide has also demonstrated its efficacy in 68 several diseases such as various inflammatory diseases including 69 rheumatoid arthritis, Crohn’s disease, and Behcet’s disease 70 [9–10]. Furthermore, thalidomide has become associated with a 71 range of immunomodulatory actions [11]. 72 In our previous studies, a series of novel isosteric thalidomide 73 analogs with a distinct and clear potent antitumor activity were 74 designed and synthesized. These thalidomide analogs lead to 75 increase of Fas-L expression, decrease in Ki67 and vascular 76 endothelial growth factor (VEGF) staining in tumor cells which 77 suggests an inhibition of tumor proliferation rate, angiogenic pro- 78 cess associated with tumor growth, decrease in lactate dehydroge- 79 nase (LDH), intracellular adhesion molecule (ICAM-1) expression http://dx.doi.org/10.1016/j.cbi.2015.05.017 0009-2797/Ó 2015 Elsevier Ireland Ltd. All rights reserved. ⇑ Corresponding author at: Faculty of Science, Department of Chemistry, Menoufia University, Shebin El-Koam 2323, Egypt. Fax: +20 24159770. E-mail address: magdyzahran53@gmail.com (M. Zahran). Chemico-Biological Interactions xxx (2015) xxx–xxx Contents lists available at ScienceDirect Chemico-Biological Interactions journal homepage: www.elsevier.com/locate/chembioint CBI 7367 No. of Pages 8, Model 5G 4 June 2015 Please cite this article in press as: R. Talaat et al., Anti-inflammatory effect of thalidomide dithiocarbamate and dithioate analogs, Chemico-Biological Inter- actions (2015), http://dx.doi.org/10.1016/j.cbi.2015.05.017