Int. J. Pharm. Investigation, 2020;10(1):37-42 International Journal of Pharmaceutical Investigation, Vol 10, Issue 1, Jan-Mar, 2020 37 Original Article Comparative Pulmonary Protective Efcacy of Amifostine and it’s Analogue S-2(2-aminoethylamino)ethyl Phenyl Sulfde (DRDE-07) against Sulphur Mustard Induced Oxidative Stress and Infammation in Female Mice Alok Kumar Soni 1 , Uma Pathak 2 , Durga Prasad Nagar 1 , Arvind Kumar Gupta 3 , Gurusamy Mathu Kannan 1, * 1 Division of Pharmacology and Toxicology, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, INDIA. 2 Division of Synthetic Chemistry, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, INDIA. 3 Process Technology Development Division, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, INDIA. ABSTRACT Aim: The present study was undertaken to investigate the comparative pul- monary protective ef fcacy of Amifostine (S-2[3-aminoprophylamino] ethyl phosphorothioate) and its analogues DRDE-07 (S-2(2-aminoethylamino) ethyl phenyl sulfde) against sulfur mustard toxicity in mice. Materials and Methods: Twenty female mice were divided into four groups: Control, SM group animals were percutaneously exposed to 16.2 mg/kg. The third and fourth group of animals received amifostine and DRDE-07 (210 and 250 mg/kg respectively) through the oral route, 30 min before SM exposure. The clinical symptoms and body weight changes were observed daily and sacrifced on 7 th day. Bronchoalveolar lavage fuid (BALF) and lung tissue were collected for biochemical and histopathological studies. The follow- ing biochemical endpoints were studied in BALF (total cell count, lactate dehydrogenase, protein content, β-glucuronidase activity, MMP-2, 9 activ- ity and FSH) whereas reactive oxygen species (ROS), reduced glutathione (GSH), lipid peroxidation, superoxide dismutase, catalase and myeloper- oxidase activity was measured in lung tissue. The above biochemical ob- servations are also supported by histopathology studies. Results: Dermal exposure to SM signifcantly reduced body weight. The signifcant increase in BALF LDH leakage, protein content, cell number and MMPs activity in the SM exposed animals suggest disruption of endothelial barrier in the lung (p<0.05). Signifcant ROS generation (p<0.05) was observed in lung tissue of SM group which results in a signifcant decrease in SOD GSH and CAT and an increase in MDA (p<0.05). These alterations in BALF as well in lung tissue due to SM exposure was signifcantly prevented by the pretreatment of amifostine and DRDE-07 (p<0.05). The histopathological observations also support the above results. The above results indicate that the preventive ef fcacy of DRDE-07 is better than amifostine. Conclusion: The percutaneous SM exposure-induced pulmonary damages were signif- cantly protected by DRDE-07 than amifostine in mice. Keywords: Amifostine, BALF, Chemical Warfare Agents, DRDE-07, Pulmo- nary injury, Oxidative stress, Sulfur mustard. Correspondence Dr. Gurusamy Mathu Kannan Division of Pharmacology and Toxicology, Defence Research and Development Es- tablishment, Jhansi Road, Gwalior - 474 002, Madhya Pradesh, INDIA. Phone no: +91-0751-2390369 Email: gmkannan@drde.drdo.in DOI: 10.5530/ijpi.2020.1.7 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms. INTRODUCTION Sulfur mustard (SM, 2, 2’-dichlorodiethyl sulfde) is a vesicant chemi- cal warfare agent which was widely used during World War-I, Iran-Iraq conficts and recently by the Syrian terrorist groups. 1 Skin, eye and re- spiratory system are the major target organs to SM exposure and induce a small blister to severe pathological alteration and even death. 2 Even single exposure to mustard gas with respiratory injury was associated with increased risk of lung cancer in later life. 3 Because of the lipophilic- ity nature, SM penetrates the human skin and distributed to the whole body within a short time, where they induced internal organs injury. Vi- jayaraghavan et al. (2001) demonstrated that SM is highly toxic through the dermal route than subcutaneous and oral route. 4 Recently, Mohamed Batal et al also reported that percutaneous SM exposure induces time and dose-dependent DNA damage in internal organs and lung was the most afected organ. 5 Oxidative stress and infammations are attributed to the SM induced pulmonary toxicity in various experimental animals. 6 Te subcutaneous injection of half mustard induced oxidative stress through free radical generation and activation of the immune system through the infltration of T cells and generation of infammatory cytokines which are responsible for toxic efects in the lungs. 7,8 Various strategies have been adopted to counteract the SM toxicities which include adminis- tration of antioxidant, anti-infammatory agents, protease inhibitors and radio protector. 9 Treatment by antioxidant and anti-infammatory agents have been reported for SM induced systemic injury include such as melatonin, pentoxifylline, N-acetyl cysteine, sodium thiosulphate. 10,11 But none of them found to be efective in preventing the SM induced lethal toxicity. Amifostine (WR-2721) is an analogue of cysteamine and selectively protect the normal tissues from the toxicities associated with chemo- therapy and irradiation. 12 Te cytoprotective properties of amifostine are attributed to the potent scavenging of drug or radiation-induced oxygen free radicals. 11 Since the SM toxicity mimics the radiation toxic- ity the prophylactic efcacy of amifostine was tested against SM both in vitro 13 and in vivo. 14 Few studies were also compared to the protective efcacy of amifostine and DRDE-07 on diferent animal models against SM and nitrogen mustard. 15,16 Tough the above studies suggested the better prophylactic efcacy of DRDE-07 than amifostine, on haemato- logical, hepatic and renal biochemical parameters, there is no report on the pulmonary protection. Te present study was aimed to investigate