Original article Anticancer activity of new coumarin substituted hydrazidee hydrazone derivatives Tamer Nasr a, * , Samir Bondock b, c , Mahmoud Youns d a Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Helwan University, Helwan, Egypt b Department of Chemistry, Faculty of Science, Mansoura University, ET-35516 Mansoura, Egypt c Department of Chemistry, Faculty of Science, King Khalid University, 9004 Abha, Saudi Arabia d Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, Helwan University, Egypt article info Article history: Received 12 November 2013 Received in revised form 24 December 2013 Accepted 8 February 2014 Available online 11 February 2014 Keywords: Coumarin Hydrazideehydrazone Antiproliferative Apoptosis Caspases 3/7 Microarray abstract Drug resistance is a major impediment for cancer treatment, to overcome it we designed and synthesized sixteen coumarins bearing hydrazideehydrazone moiety and evaluated them against human drug-resistant pancreatic carcinoma (Panc-1) cells and drug-sensitive (hepatic carcinoma; Hep-G2 and leukemia; CCRF) cell lines in vitro. The 6-brominated coumarin hydrazideehydrazone derivatives (BCHHD) 7c, 8c and 10c were more potent than doxorubicin (DOX) against resistant Panc-1 cells. BCHHD 7c showed significant cytotoxicity against all tested cells (IC 50 : 3.60e6.50 mM) on comparison with all other coumarin hydrazideehydrazone derivatives (CHHD), whereas BCHHD’s 8c and 10c showed sig- nificant antiproliferative activity only against resistant Panc-1 cells with IC 50 of 2.02 mM and 2.15 mM, respectively. All the investigated BCHHD’s were able to activate caspases 3/7 and they could induce apoptosis in resistant Panc-1 cells. Microarray analysis showed that BCHHD 7c induced the expression of apoptotic- and cell cycle arrest (G2/M)- genes in resistant Panc-1 cells. Moreover, BCHHD 7c induced the up-regulation of CDKN1A, DDIT4, GDF-15 and down-regulation of CDC2, CDC20, CDK2 genes. Based on our results, we conclude that 7c could be a potent anticancer drug to overcome drug resistance in cancer and it could be highly beneficial for patients in the clinic. Ó 2014 Elsevier Masson SAS. All rights reserved. 1. Introduction Drug resistance constitutes lack of response to many chemically and mechanically unrelated anticancer agents by cancer cells. It is one of the main causes for failure of chemotherapy and can lead to recurrence of disease or even death [1]. Clinical administration of high doses of anticancer drugs to overcome resistance leads to drug-induced toxicities [2]. Hence newer anticancer agents need to be synthesized and tested for its efficacy both in vitro and in vivo to overcome drug resistance. The natural and synthetic coumarins attract great attention due to their wide range of biological properties, including anticancer [3], anti-HIV [4], anti-inflammatory [5] and antibacterial [6] activ- ities. Furthermore, their cancer chemopreventive properties have been recently emphasized [3,7]. The apoptosis and differentiation- induced activities of coumarins extend to several different cell line models in vitro, and they appear to be the most promising in terms of cancer treatment [8]. Coumarins could exert their anticancer activity by different mechanisms; either by inhibiting the telomerase enzyme [9], inhibiting protein kinase activity and down regulating oncogene expression [10] or by inducing the caspase-9 mediated apoptosis. Additionally, researchers showed that coumarins are able to sup- press cancer cell proliferation by arresting cell cycle in G0/G1 [9], G2/M phases [11], and through affecting the p-gp of the cancer cells [12,13]. It was also reported that hydroxycoumarins might exert their anticancer activity by generating free radical species in cancer cells producing oxidative stress leading to pro-apoptic effect [7]. It was proven that the d-lactone ring of the coumarinic system has a fundamental role in both the generation and stabilization of such species as well as in the pro-apoptotic action of hydroxycoumarins [7]. Moreover, the antiproliferative activity of 7-hydroxycoumarin derivatives could be due to their effect on the mitochondrial thiol compounds of cancer cells [14]. Literature survey revealed that the hydrazideehydrazone (eCOeNHeN]CHe) moiety has significant role as antitumor * Corresponding author. E-mail address: tamerhefni@yahoo.com (T. Nasr). Contents lists available at ScienceDirect European Journal of Medicinal Chemistry journal homepage: http://www.elsevier.com/locate/ejmech http://dx.doi.org/10.1016/j.ejmech.2014.02.026 0223-5234/Ó 2014 Elsevier Masson SAS. All rights reserved. European Journal of Medicinal Chemistry 76 (2014) 539e548