In silico study, synthesis, and in vitro cytotoxic activity of novel 2-mercaptobenzimidazole derivatives

Year 2025, Volume: 29 Issue: 6, 2402 - 2418, 02.11.2025

Shams A. Nadhum Mohammed Hassan Mohammed

https://doi.org/10.12991/jrespharm.1797989

Abstract

Based on the benzimidazole molecule, two new series of 2-mercaptobenzimidazole derivatives contain different substitutions one at the mercapto (SH) group (different amines) and the second at N1 of the benzimidzole ring (different phenyl moiety) were synthesized in this research. The structures of the synthesized compounds were verified utilizing mass spectroscopy, fundamental analysis data, 1H-NMR, 13C-NMR, and FT-IR spectroscopic methods. The cytotoxicity efficacy of the target drugs were assessed using MTT assays and their interactions with the target enzymes of the human oestrogen and the epidermal growth factor receptors were predicted using molecular docking. Compound 6C demonstrated a very good cytotoxicity result against MCF-7 cancer cells with an IC50 of 62.20 µM, which is comparable to the standard 4-hydroxytamoxifen, which has an IC50 of 70.27 µM. Compound 2C demonstrated very good effects against A549 cancer cells, which has an IC50 of 72.28 µM, which is comparable to the standard gefitinib, which has an IC50 of 75.10 µM, and the docking results show compound 2C has an energy of binding of -7.26 Kcal/mol, which is comparable to the standard gefitinib, which has an energy of -7.69 Kcal/mol, while compound 2C and 3C have energy of -9.23,-8.94 Kcal/mol, respectively, which is comparable to the standard 4-hydroxytamoxifen, which has a binding energy of -9.11 Kcal/mol. Also, computational programmes were used to determine the compounds' drug-like properties, showing that all derivatives had good results. These findings propose that these inhibitors may be helpful in the progression of new anticancer medications in the future.

Keywords

Cytotoxic agents , 2- mercaptobenzimidazole , molecular docking , epidermal growth factor receptor

References

• [1] Oleiwi MA, Zalzala MH. Synthesis, molecular docking study and cytotoxicity evaluation of some quinazolinone derivatives as nonclassical antifolates and potential cytotoxic agents. Iraqi J Pharm Sci. 2022; 31(2): 283–296. https://doi.org/10.31351/vol31iss2pp283-296
• [2] Nadhum SA, Mohammed MH. Design, synthesis, characterization and preliminary anticancer study for methotrexate silibinin conjugates. Iraqi J Pharm Sci. 2015; 24(1): 74–84. https://doi.org/10.31351/vol24iss1pp74-84
• [3] Saeed AM, Al-Hamashi AA. Molecular docking, ADMET study, synthesis, characterization, and preliminary antiproliferative activity of potential histone deacetylase inhibitors with isoxazole as new zinc binding group. Iraqi J Pharm Sci. 2023; 32(Suppl.): 188–203. https://doi.org/10.31351/vol32issSuppl.pp188-203
• [4] Hassan OM, Mahmood AA, Tahtamouni L. Synthesis and docking studies of new 5-bromoindole-2-carboxylic acid oxadiazole derivatives as EGFR inhibitors. Tikrit J Pharm Sci. 2023; 17(1): 58–77. http://doi.org/10.25130/tjphs.2023.17.1.6.58.77
• [5] Torres RJ, José ES. Src tyrosine kinase inhibitors: New perspectives on their immune, antiviral, and senotherapeutic potential. Front Pharmacol. 2019; 10: 1011–1017. https://doi.org/10.3389/fphar.2019.01011
• [6] Al-khfajy WS, Zalzala MH, Al-Mashhadani Z. Synergistic effects of 2-deoxy-D-glucose and cinnamic acid with erlotinib on NSCLC cell line. Iraqi J Pharm Sci. 2023; 32(1): 136–144. https://doi.org/10.31351/vol32issSuppl.pp136-144
• [7] Yaseen YS, Mahmood AA, Abbas AH, Shihab WA, Tahtamouni LH. New niflumic acid derivatives as EGFR inhibitors: Design, synthesis, in silico studies, and anti-proliferative assessment. Med Chem. 2023; 19: 445–459. https://doi.org/10.2174/1573406419666221219144804
• [8] Fan M, Yao L. Design, synthesis, and in vitro antitumor activity of 6-aryloxyl substituted quinazoline derivatives. Turk J Chem. 2022; 46: 849–858. https://doi.org/10.55730/1300-0527.3373
• [9] Hassan OM, Mahmood AA, Tahtamouni L. Novel 5-bromoindole-2-carboxylic acid derivatives as EGFR inhibitors: synthesis, docking study, and structure-activity relationship. Anti-Cancer Agents Med Chem. 2023; 23(11): 1336– 1348. https://doi.org/10.2174/1871520623666230227153449
• [10] Zhang Y, Wang Q. Synthesis and preliminary structure-activity relationship study of 3-methylquinazolinone derivatives as EGFR inhibitors with enhanced antiproliferative activities against tumour cells. J Enzyme Inh Med Chem. 2021; 36(1): 1205–1216. https://doi.org/10.1080/14756366.2021.1933466
• [11] Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer. Mol Oncol. 2018; 12: 3–20. https://doi.org/10.1002/1878-0261.12155
• [12] Lu X, Yu L, Zhang Z, Ren X, Smaill JB, Ding K. Targeting EGFR(L858R/T790M) and EGFR(L858R/T790M/C797S) resistance mutations in NSCLC: Current developments in medicinal chemistry. Med Res Rev. 2018; 38(5): 1550– 1581. https://doi.org/10.1002/med.21488
• [13] Pashayan N, Antoniou AC, Ivanus U, Esserman LJ, Easton DF Garcia-Closas M, Schmutzler R, Wegwarth O, Pharoah P, Moorthie S, De Montgolfier S, Baron C, Herceg Z, Turnbull C, Balleyguier C, Rossi PG, Wesseling J. Personalized early detection and prevention of breast cancer: Envision consensus statement. Nat Rev Clin Oncol. 2020; 17(11): 687–705. http://doi.org/10.1038/s41571-020-0388-9
• [14] Sukocheva OA. Estrogen, estrogen receptors, and hepatocellular carcinoma: Are we there yet? World J Gastroenterol. 2018; 24(1): 1–4. https://doi.org/10.3748/wjg.v24.i1.1
• [15] Guillaume M, Montagner A, Fontaine C, Lenfant F, Arnal JF, Gourdy P. Nuclear and membrane actions of estrogen receptor alpha: Contribution to the regulation of energy and glucose homeostasis. Adv Exp Med Biol. 2017; 1043: 401–426. https://doi.org/10.1007/978-3-319-70178-3_19
• [16] Kucukoglu K, Secıntı H, Ozgur A, Secen H, Tutar Y. Synthesis, molecular docking, and antitumoral activity of alnustone-like compounds against estrogen receptor alpha-positive human breast cancer. Turk J Chem. 2015; 39(1): 179–193. https://doi.org/10.3906/kim-1408-72
• [17] Nose N, Sugio K, Oyama T, Nozoe T, Uramoto H, Iwata T, Onitsuka T, Yasumoto K. Association between estrogen receptor beta expression and epidermal growth factor receptor mutation in the postoperative prognosis of adenocarcinoma of the lung. J Clin Oncol. 2009; 27(3): 411–417. https://doi.org/10.1200/jco.2008.18.3251
• [18] Stabile LP, Lyker JS, Gubish CT, Zhang W, Grandis JR, Siegfried JM. Combined targeting of the estrogen receptor and the epidermal growth factor receptor in non-small cell lung cancer shows enhanced antiproliferative effects. Cancer Res. 2005; 65(4): 1459–1470. https://doi.org/10.1158/0008-5472.can-04-1872
• [19] Nadhum SA, Kamoon RA, Mohammed MH. 6-Mercaptopurine derivatives : Maintenance therapy of acute lymphoblastic leukemia: A review. Biochem Cell Arch. 2020; 20(1): 2091–2099. https://doi.org/10.35124/bca.2020.20.1.2091
• [20] Abduljabbar TT, Hadi MK. Synthesis, characterization and antibacterial evaluation of some coumarin derivatives. Iraqi J Pharm Sci. 2021; 30(1): 249–257. https://doi.org/10.31351/vol30iss1pp249-257
• [21] Kandemir L, Karakuş S, Özbaş S, Rollas S, Akbuğa J, Kandemir L. Synthesis, structure elucidation and cytotoxic activities of 2,5-disubstituted-1,3,4-thiadiazole and 1,2,4-triazole-3-thione derivatives. J Res Pharm. 2022; 26(4): 941– 953. https://doi.org/10.29228/jrp.192
• [22] Mohareb RM, Gamaan MS. The uses of ethyl 2-(1h-benzo[d]imidazol-2-yl)acetate to synthesis pyrazole, thiophene, pyridine and coumarin derivatives with antitumor activities. Bull Chem Soc Ethiop. 2018; 32(3): 541–557. https://doi.org/10.4314/bcse.v32i3.13
• [23] Al-Majidi SM, Ibrahim HA, AL-issa YA. Synthesis and identification of some new derivatives [benzyl thio]benzimidazole-N-(methylene-5-yl) 4,5-di substituted 1,2,4-triazole and evaluation of their activity as antimicrobial and anti-inflammatory agents. Iraqi J Sci. 2021; 62(4): 1054–1065. https://doi.org/10.24996/IJS.2021.62.4.2
• [24] Kalay Ş, Akkaya H. Molecular modelling of some ligands against acetylcholinesterase to treat Alzheimer’s Disease. J Res Pharm. 2023; 27(6): 2199-2209. http://doi.org/10.29228/jrp.510
• [25] Özdemir Z, Alagöz MA, Akdemir AG, Özçelik AB, Özçelik B, Uysal M. Studies on a novel series of 3(2H)- pyridazinones: Synthesis, molecular modelling, antimicrobial activity. J Res Pharm. 2019; 23(5): 960-972. https://doi.org/10.35333/jrp.2019.43
• [26] Wodtke R, Steinberg J, Kockerling M, Loser R, Mamat C. NMR-based investigations of acyl-functionalized piperazines concerning their conformational behaviour in solution. RSC Adv. 2018, 8: 40921–40933. http://doi.org/10.1039/C8RA09152H
• [27] Murtaza S, Altaf AA, Hamayun M, Iftikhar K, Tahir MN, Tariq J, Faiz KH .Synthesis, antibacterial activity and docking studies of chloroacetamide derivatives. Eur J Chem. 2019; 10(4): 358-366. http://doi.org/10.5155/eurjchem.10.4.358-366.1859
• [28] Yadav S, Narasimhan B, Lim SM, Ramasamy K. Synthesis and evaluation of antimicrobial, antitubercular and anticancer activities of benzimidazole derivatives. Egypt J Basic Appl Sci. 2018; 5(1): 100–109. https://doi.org/10.1016/j.ejbas.2017.11.001
• [29] Frick M, McAtee D, McAtee J. Synthesis of N -alkyl-1,4-diazepin-5-ones via Schmidt ring expansion chemistry. Heterocycl Commun. 2011; 17(1-2): 17–19. http://doi.org/10.1515/hc.2011.002
• [30] Shihab IK, Mohammed MH. Synthesis, characterization and preliminary cytotoxic activity study of new 5- fluorouracil conjugate with pyrrolidine dithiocarbamate as a mutual anticancer prodrug. Iraqi J Pharm Sci. 2019;28(2):17–23. https://doi.org/CorpusID:210895103
• [31] Çimen Z, Akkoç S, Kökbudak Z. Reactions of aminopyrimidine derivatives with chloroacetyl and isophthaloyl chlorides. Heteroatom Chem. 2018;29:1-6. https://doi.org/10.1002/hc.21458
• [32] Al-kazweeny R, Muhi-eldeen ZA, Al-kaissi E, Al-tameemi S, Tayeh SS, Al-hussenini J. Designs, synthesis, structural elucidation and antimicrobial evaluation of various derivatives of 2-mercaptobenzimidazole as possible antimicrobial agents. Int J Med Res Health Sci. 2020; 9(12): 1-13. https://doi.org/CorpusID:233366853
• [33] Molou KYG, Timotou A, Camara TE. Design, synthesis of a novel N-substituted benzimidazole derivatives. Acta Chim Pharm Indica. 2022;12(3):1-7. http://doi.org/10.37532/2277-288X.2022.12(3).184
• [34] Abdullah MN, Hamid SA, Salhimi SM, Jalil NA, Al-Amin M, Jumali NS. Design and synthesis of 1 sec/tert butyl-2- chloro/nitrophenyl benzimidazole derivatives: Molecular docking and in vitro evaluation against MDA-MB-231 and MCF-7 cell lines. J Mol Struct. 2023; 1277:13428. https://doi.org/10.1016/j.molstruc.2022.134828
• [35] Priimagi A, Cavallo G, Metrangolo P, Resnati G. The halogen bond in the design of functional supramolecular materials: Recent advances. Acc Chem Res. 2013; 46(11): 2686–2695. https://doi.org/10.1021/ar400103r
• [36] Elhemely MA, Belgath AA, El-Sayed SH, Burusco KK, Kadirvel M, Tirella A, Finegan K, Bryce RB, Stratford IJ, Sally Freeman S. SAR of novel 3-arylisoquinolinones: meta-Substitution on the aryl ring dramatically enhances antiproliferative activity through binding to microtubules. J Med Chem. 2022; 65( 6): 4783–4797. https://doi.org/10.1021/acs.jmedchem.1c01936
• [37] Cheng M, Yu X, Lu K, Xie L, Wang L, Meng F, Han X, Chen X, Liu J, Xiong Y, Jin J. Discovery of potent and selective epidermal growth factor receptor (EGFR) bifunctional small-molecule degraders. J Med Chem. 2020; 63: 1216−1232. https://doi.org/10.1021/acs.jmedchem.9b01566
• [38] Todsaporn D, Mahalapbutr P, Poo-Arporn RP, Choowongkomon K, Rungrotmongkol T. Structural dynamics and kinase inhibitory activity of three generations of tyrosine kinase inhibitors against wild-type, L858R/T790M, and L858R/T790M/C797S forms of EGFR. Comput Biol Med. 2022; 147: 105787–105799. https://doi.org/10.1016/j.compbiomed.2022.105787
• [39] Ervina M, Pratama MR, Poerwono H, Ekowati J, Widyowati R, Matsunami K, Sukardiman. In silico estrogen receptor alpha antagonist studies and toxicity prediction of Melia azedarach leaves bioactive ethyl acetate fraction. J Adv Pharm Technol Res. 2021; 12(3): 236–241. https://doi.org/10.4103/japtr.japtr_198_21
• [40] Sagheer OM, Mohammed MH, Wadi JS, Ibraheem ZO. Studying the cytotoxic activity of newly designed and synthesized HDAC inhibitors derivatives of pentanoyl anilide-5-biguanide. Macromol Symp. 2022; 401(1): 1–10. https://doi.org/10.1002/masy.202100346
• [41] Sulaiman GM, Jabir MS, Hameed AH. Nanoscale modification of chrysin for improved of therapeutic efficiency and cytotoxicity. Artif Cells Nanomed Biotechnol. 2018; 46(sup1): 708–720. https://doi.org/10.1080/21691401.2018.1434661