Contents lists available at ScienceDirect Bioorganic Chemistry journal homepage: www.elsevier.com/locate/bioorg Design and synthesis of novel 2,3-dihydropyrazino[1,2-a]indole-1,4-dione derivatives as antiproliferative EGFR and BRAF V600E dual inhibitors Lamya H. Al-Wahaibi a , Ahmed M. Gouda b , Ola F. Abou-Ghadir c , Ola I.A. Salem c , Asmaa T. Ali d , Hatem S. Farghaly d , Mostafa H. Abdelrahman e , Laurent Trembleau f , Hajjaj H.M. Abdu-Allah c, ⁎ , Bahaa G.M. Youssif c, ⁎ a Department of Chemistry, College of Sciences, Princess Nourah Bint Abdulrahman University b Department of Medicinal Chemistry, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62514, Egypt c Pharmaceutical Organic Chemistry Department, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt d Biochemistry Department, Faculty of Pharmacy, Nahda University, Beni-Suef, Egypt e Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Al-Azhar University, Assiut 71524, Egypt f School of Natural and Computing Sciences, University of Aberdeen, Meston Building, Aberdeen AB243UE, United Kingdom ARTICLE INFO Keywords: Pyrazino-indole Antiproliferative EGFR BRAF V600E Apoptosis ABSTRACT Recent studies have shown additive and synergistic efects associated with the combination of kinase inhibitors. BRAF V600E and EGFR are attractive targets for many diseases treatments and have been studied extensively. In keeping with our interest in developing anticancer targeting EGFR and BRAF V600E , a novel series of 2,3-dihy- dropyrazino[1,2-a]indole-1,4-dione has been rationally designed, synthesized and evaluated for their anti- proliferative activity against a panel of four human cancer cell lines. Compounds 20–23, 28–31, and 33 showed promising antiproliferative activities. These compounds were further tested for their inhibitory potencies against EGFR and BRAF V600E kinases with erlotinib as a reference drug. Compounds 23 and 33 exhibited equipotency to doxorubicin against the four cell lines and efciently inhibited both EGFR (IC 50 = 0.08 and 0.09 µM, respec- tively) and BRAF V600E (IC 50 = 0.1 and 0.29 µM, respectively). In cell cycle study of MCF-7 cell line, compounds 23 and 33 induced apoptosis and exhibited cell cycle arrest in both Pre-G1 and G2/M phases. Molecular docking analyses revealed that the new compounds can ft snugly into the active sites of EGFR, and BRAF V600E kinases. Compound 23, 31 and 33 adopted similar binding orientations and interactions to those of erlotinib and ve- murafenib. 1. Introduction Because several kinases are involved in the growth of human can- cers, protein kinases have emerged as a major class of drug therapy targets [1,2]. Up to now, approximately 50 kinase inhibitors (KIIs) have already been licensed by the US Food and Drug Administration (FDA), among them, 45 kinase inhibitors are currently being targeted for cancer treatment [3]. As a result, much efort has been made to es- tablish small-molecule inhibitors that interact directly with the cata- lytic activity of certain target protein kinases. While usually established against single or more targets, small molecule protein kinase inhibitors often have a signifcant cross-reactivity as they can target preserved structural components, including protein kinases with purine nucleo- tide binding pockets [4]. One-target treatment has recently been widely reported to result in resistance to chemotherapy [5]. To fnd a solution for single-target drug treatment for chemotherapeutic cancer, combi- nation therapy has recently been adopted for clinical use [6]. Combi- nation therapy can have potential additive and even synergistic efects, but often results in some unexpected side-efects, such as an increased toxicity. As an alternative approach for combination therapy, dual- target or multi-target drugs have a lower hazard of drug interactions, improved pharmacokinetics (PK) and safety profles. A dual-target ki- nase can also prevent two drug interactions, poor patient adherence, adverse of-target efects, and high production costs [7]. There are many FDA approved dual-target or multi-target anticancer drugs. Alectinib is ALK/RET dual-targeted drug for ALK NSCLC [8]. Dasa- tinib has nine known targets (BCR-Abl, EGFR, Src, Lck, Yes, Fyn, Kit, EphA2, PDGFR) for CML or ALL. Briefy, dual/multi-targeted kinase inhibitors are well adapted to the development of efective anticancer drugs [9]. https://doi.org/10.1016/j.bioorg.2020.104260 Received 27 June 2020; Received in revised form 13 August 2020; Accepted 28 August 2020 ⁎ Corresponding authors at: Pharmaceutical Organic Chemistry Department, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt. E-mail addresses: hajjaj@aun.edu.eg (H.H.M. Abdu-Allah), bgyoussif@ju.edu.sa, bahaa.youssif@pharm.aun.edu.eg (B.G.M. Youssif). Bioorganic Chemistry 104 (2020) 104260 Available online 03 September 2020 0045-2068/ © 2020 Elsevier Inc. All rights reserved. T