Synthesis, and  Anti-Tumor Evaluation of Some New Flurbiprofen  Derivatives Against MCF-7  and WRL-68 Cell Lines

Kasim S. Hmood; Ammar A. Razzak  Mahmood  Kubba; Redha  I Al-bayati; Abdulrahman M.  Saleh

doi:10.22146/ijp.730

Kasim S. Hmood Iraqi Ministry of Health, Wasit Health Directorate- Department of Pharmacy, Wasit-Iraq
Ammar A. Razzak Mahmood Kubba Department of Pharmaceutical Chemistry, College of Pharmacy-University of Baghdad, Baghdad, Bab-Al-Mouadam-10001-Iraq
Redha I Al-bayati Department of Chemistry, College of Science, Al-Mustansirya University, Baghdad, 10001, Iraq
Abdulrahman M. Saleh Pharmaceutical Medicinal Chemistry and Drug Design Department, Faculty of Pharmacy (Boys), Al-Azhar University. Cairo 11884, Egypt

Keywords: Thiosemicarbazide, anticancer activity, thiourea, aryl isothiocyanate, high content screening

Abstract

A new series of flurbiprofen derivatives containing thiosemicarbazide moiety (3-7) was synthesized from flurbiprofen as parent nucleus by esterification, hydrazide formation, and heating with different aryl isothiocyanate substituents, respectively. Flurbiprofen was also treated with thiosemicarbazide in the presence POCl₃ as a catalyst, to produce 1,3,4 -thiadiazole -2-amine (8). Treatment of (8) with different aryl isothiocyanates produced thiourea derivatives (9-12). Also, the reaction of (8) with different benzoyl chloride substituents produced benzamide compounds (13-15). Eventually , treatment of (8) with ethyl acetoacetate(EAA) produced [1,3,4]thiadiazolo[3,2-a]pyrimidin-7-one (16) .The new compounds were characterized by spectroscopic techniques :FTIR, ¹HNMR, and CHNS analysis. A molecular docking study for the synthesized compounds (3-16), against the Vascular Endothelial Growth Factor receptor (VEGFR-2) was applied and it indicated that compounds 4,7,13, and 15,exhibited the optimum binding energy of -6.77, -6.12,-6.68, and -6.43 kcal/mol, respectively. Target compounds were also assessed for their in vitro anticancer effects in a cell-line study. All of the compounds tested showed the most plausible anticancer activity, compared to a positive control(Sorafenib), using in vitro MTT cytotoxic assay ,against human breast tumor (MCF-7), and normal WRL-68 cell line. The in vitro results revealed that compounds 4,5,10,11,13, and 15 exhibited the highest inhibitory activity at their IC₅₀ concentrations, against MCF-7 cell lines, as follows (122.7,113.9,95,7. 109.1,40.32 and 112.29µg/mL, respectively. While their cytotoxic effect against normal WRL-68 cell line at their IC₅₀ concentrations, as follow 210.2, 181.3 ,151.7,278.7,80.28, and 236 µg/mL, respectively, therefore, such compounds were considered more selective toward MCF-7 than normal WRL-68,and their selectivity index (SI): 1.71,1.59,1.59 ,2.55 ,1.99 , and 2.10,respectively . Among the synthesized compounds, the compound 15 was chosen to screen its effect in vitro through multi-parameter cytotoxic assay against MCF-7 breast cancer implemented in High Content Screening (ArrayScan XTI, Thermo Scientific),which could be taken in consideration as a starting point for the development of new anticancer drugs

References

Abdel Rahman ED., Mohamed KO. 2014. Synthesis of novel 1,3,4-thiadiazole analogues with expected anticancer activity. Der Pharma Chemica. 6(1) 323-335, http://derpharmachemica.com/archive.html.

Abraham V.C.,Towne D.L., Waring J.F., Warrior U., Burns D.J. 2008. Application of a high-content multiparameter cytotoxicity assay to prioritize compounds based on toxicity potential in humans. J Biomol Screen.13(6)527-537, doi: 10.1177/1087057108318428.

Al-Saad HN., Kubba AAR, Al-Bayati I.R. 2019. Design, synthesis, docking study and antiplatelet evaluation of new thiosemicarbazide derivatives, derived From captopril. Orient J Chem.,35(2) 829-838,http://dx.doi.org/10.13005/ojc/350246.

Al-Saad HN., Kubba AAR.2020. Evaluation of new thiosemicarbazides derived from captopril as angiotensin converting- enzyme inhibitors with docking study and predicted ADMET analysis. Res J Pharm Tech.,13(6) 2733-2741, doi: 10.5958/0974-360X.2020.00486.2.
Al-Wabli RI., Zakaria AS., Attia MI. 2017.Synthesis, spectroscopic characterization and, antimicrobial potential of certain new isatin-indole molecular hybrids, Molecules, 22(11)1958-1974, doi: 10.3390/molecules22111958.
Arora S., Agarwal S., Singhal S. 2014 . Anticancer activities of thiosemicarbazide / thiosemicarbazones: A review, Int J Pharm Pharm Sci.,6(9)34-41
Bhattacharya P., Leonard J.T., Roy K. 2005. Exploring QSAR of thiazole and thiadiazole derivatives as potent and selective human adenosine A3 receptor antagonists using FA and GFA techniques. Bioorg Med Chem.,13(4)1159–116, http:// doi: 10.1016/j.bmc.2004.11.022.

Brooks BR. , Brooks CL., MacKerell AD., Nilsson L., Petrella RJ., Roux B.,Won Y., Archontis G.,Bartels C.,Bor S. 2009. CHARMM: the biomolecular simulation program. J Comput Chem.,30(10)1545-1614, https://doi.org/10.1002/jcc.21287.

Chan JOT , Arsianti A, Marcelia M., Wijoyo SJ., Fadilah F., Putrianingsih R. ,Azizah NN. ,Tanimoto H. , Arsianti KK. 2018. Synthesis and anticancer effect of 3,4,5-N-alkyl-benzamides on colon carcinoma HCT- 116 Cells, Orient J Chem., 34(3)1362-1367, http://doi:dx.doi.org/10.13005/ojc/340323.
Choo H.,Kholmukhamedov A.,Zhou C.,Jobe S. 2017. Inner mitochondrial membrane disruption links apoptotic and a Agonist-initiated phosphatidylserine externalization in platelets, Arterioscler Thromb Vasc Biol., 37(8)1503–1512, doi: 10.1161/ATVBAHA.117.309473.

Coutsias EA.,Seok C.,Dill K.. 2004. Using quaternions to calculate RMSD. J Comput Chem.,25(15): 1849-1857, doi: 10.1002/jcc.20110.
El-Adl K., El-Helby A A.,Sakr H, Eissa IH., El-Hddad SSA, Shoman F MIA. 2020. Design, synthesis, molecular docking and anticancer evaluations of 5-benzylidenethiazolidine-2,4-dione derivatives targeting VEGFR-2 enzyme. Bioorg Chem.,102()104059, https://doi.org/10.1016/j.bioorg.2020.104059.

Eissa I.H., El-Naggar A.M. , Abd El-Sattar NEA., Youssef ASA. 2018. Design and discovery of novel quinoxaline derivatives as dual DNA intercalators and topoisomerase II inhibitors, Bentham science, 18(2) 195-209, doi: 10.2174/1871520617666170710182405.

El-Far M., Elmegeed G.A., Eskander E.F. 2009. Rady H.M , Novel modifed steroid derivatives of androstanolone as chemotherapeutic anti-cancer agents. Eur J Med Chem., 44 (10) 3936–3946, doi: 10.1016/j.ejmech.2009.04.020.

El-Feky SAH., Abd El-Fattah HA.,Osman NA.,Imran M ,Zedan MN. 2015. Design, synthesis and in vitro antitumor activity of novel phthalazin-1, 4-dione/chalcone hybrids and phthalazin-1, 4-dione/pyrazoline hybrids. J Chem Pharm Res., 7(7)1154-1166.
Foroumadi A.,Kargar Z., Sakhteman A., Sharifzadeh Z.,Feyzmohammadi R., Kazemi M, Shafiee A. 2006. Synthesis and antimycobacterial activity of some alkyl[5-(nitroaryl)-1,3,4-thiadiazol-2-ylthio]propionates. Bioorg Med Chem Lett.,16(5)1164–1167, http:// doi:10.1016/j.bmcl.2005.11.087.
Gomha SM., Adel-aziz HM. 2015. Synthesis and antitumor activity of 1,3,4-thiadiazole derivatives bearing coumarine ring. Heterocycles. 91(3) 583–592, http:// doi: 10.3987/COM-14-13146.

Gomha SM., Salah TA.,Abdelhamid AO.2015. Synthesis, characterization, and pharmacological evaluation of some novel thiadiazoles and thiazoles incorporating pyrazole moiety as anticancer agents. Monatsh Chem., 146 (1)149–158, http://doi:10.1007/s00706-014-1303-09.

Hamed K. ,Syam M.,.Soheil ZM, Mehran F.,Mahboubeh R, Aditya A, Behnam K., Hapipah MA, and Mohamad IN. 2014 .“Tanacetum polycephalum (L.) Schultz- Bip . induces mitochondrial-mediated apoptosis and inhibits migration and invasion in MCF7 cells,” Molecules. 19(7) 9479–9500, doi: 10.3390/molecules19079478.

Hassan F. ,Mohammed G. ,El-Hiti GA.,Alshanon A.,Yousif E. 2018. Cytotoxic effects of tamoxifen in breast cancer cells .Unexplored Med Data., 3(3)1-9, doi: 10.20517/2572-8180.2017.25.
Hmood KS., Kubba A.A.R., Synthesis, Docking study and in vitro anticancer evaluation of new derivatives of 2-(1-(2-flouro-[1,1-biphenyl]-4-yl)ethyl)-6-(substituted phenyl )imidazole[2,1-b][1,3,4]thiadiazole derived from flurbiprofen (process of publication-2020).

Huangab R.,Zhanga B.,Huang XC.,Lianga GB., Qina JM., Pana YM, Liao ZX., Wang HS. 2017. Synthesis and biological evaluation of terminal functionalized thiourea-containing dipeptides as antitumor agents RSC Adv.,7(15) 8866-8878, http:// doi:10.1039/C6RA25590F.

Hui W.,Yulan Z., Youhong H.,Shaozu W..1994. Synthesis of 5-ferrocenyl-4-phenyl-4H-1,2,4-triazole-3-thiol and Its derivatives. Synth React Inorg Met Org Chem., 24 (7)1121-1125, https://doi.org/10.1080/00945719408001388.

Issaca Y A. , Mohamedb SK. ,. Eissaa AEMF, Tantawya AH. ,El-Sawya AA. 2012. Synthesis of pentadecanyl-amino thiadiazole pharmacophores and their antimicrobial assessments. J Chem Pharm Res., 4(5) 2744-2750.

Jain K, Sharma S., Vaidya A, Ravichandran V., Agrawal RK. 2013. 1,3,4-Thiadiazole and its derivatives: A review on recent progress in biological activities. Chem. Biol. Drug Des., 81(5)557–576, http:// doi: 10.1111/cbdd.12125.

Koopaeia MN., Almasirada MJAA., Ghasemi-Nirib SF. 2013. Mohsen A.K.b,Koopaeib NN., Ghadimia M and Tabeia A., Synthesis and analgesic activity of novel hydrazide and hydrazine derivatives. Iran J Pharm Res.,12 (4) 721-727, PMID: 24523751

Kumar D., Kumar NM.,Chang KH.,Shah K. 2010. Synthesis and anticancer activity of 5-(3-indolyl)-1,3,4-thiadiazoles. Eur J Med Chem., 45(10)4664– 4668, https://doi.org/10.1016/j.ejmech.2010.07.023.

Kushwaha N.,Kushwaha SKS., Rai AK. 2012. Biological activities of thiadiazole derivatives: A review. Int J Chem Res., 4(2)517–531.

Looi C.Y.,Moharram B,Paydar M., Wong YL., Leong K.H. ,Mohamad K., Arya A., Wong W.F,Mustafa MR.2013. Induction of apoptosis in melanoma A375 cells by a chloroform fraction of Centratherum anthelminticum (L.) seeds involves NF-kappaB, p53 and Bcl-2-controlled mitochondrial signaling pathways. BMC Complement Altern Med., 13(1)166-180, http://www.biomedcentral.com/1472-6882/13/166.

Loong HH.,Yeo W. 2014.Microtubule-targeting agents in oncology and therapeutic potential in hepatocellular carcinoma. Onco Targets Ther., 7(1)575–585, doi: 10.2147/OTT.S46019.
Magalhaes LG., Ferreira L. , Andricopulo AD. 2018. Recent advances and perspectives in cancer drug design, An Acad Bras Cienc.,90(1) 1233-1250, http:// doi: 10.1590/0001-3765201820170823.

Mathew V. ,Keshavayya J., Vaidya VP., Giles D. 2007. Studies on synthesis and pharmacological activities of 3,6-disubstituted-1,2,4-triazolo[3,4-b]-1,3,4-thiadiazoles and their dihydro analogues. Eur J Med Chem., 42(6)823–840, http:// doi: 10.1016/j.ejmech.2006.12.010.

Nahi R.J.,Imran NH. 2019.Synthesis, characterization and thermal stability study of new heterocyclic compounds containing 1,2,3-triazole and 1,3,4-thiadiazole rings. Orient J Chem., 35(1) 234-240, http://dx.doi.org/ 10.13005/ojc/350128

Nishida N., Yano H., Nishida T., Kamura T., Kojiro M. 2006. Angiogenesis in Cancer, Vasc Health Risk Manag. 2(3)213–219, http:// doi: 10.2147/vhrm.2006.2.3.213.
Parmar K. ,Patel R., Joshi S.,Patel R. 2011. Efficient Synthesis and biological evaluation of 3-(phenyl)-6-(4-amino phenyl)[1,2,4]triazolo[3,4-b][1,3,4] thiadiazole and their schiff base derivatives. Int J Chemtech Res.,3(2) 761-765.

Paydar MJ, Kamalidehghan B., Wong LY., Wong WF , Looi CY, Mustafa MR. 2014. Evaluation of cytotoxic and chemotherapeutic properties of boldine in breast cancer using in vitro and in vivo models. Drug Des Devel Ther., 8:719–733, doi: 10.2147/DDDT.S58178.

Riyadh SM., El-Motairi SA., Ahmed HEA., Khalil KD., Habibe EE. 2018. Synthesis, biological evaluation, and molecular docking of novel thiazoles and [1,3,4]thiadiazoles incorporating sulfonamide group as DHFR Inhibitors. Chem. Biodivers. 15(9), http:// doi: 10.1002/cbdv.201800231.

Rudraraju A.V.,Amoyaw PNA., Hubin TJ., Khan MOF. 2014.Determination of log p values of new cyclen based antimalarial drug leads using RP-HPLC. Arch Pharm., 69(9) 655-662, doi: 10.1691/ph.2014.4019.

Sadeghinia A. , Kahroba H., Hamid A.S., Heidari R., Bradaran B., Zeinali S., Molavi O. 2019.Therapeutic targeting of angiogenesis molecular pathways in angiogenesis-dependent diseases, Biomed PharmacoTher., 110 : 775-785, http:// doi:10.1016/j.biopha.2018.12.022.

Sebeka AAH.,El Bahanasawy M, Tantawy M.A., Osman AMA., El Sayed IET. 2017. Synthesis and antiproliferative activity of novel neocryptolepine-hydrazides hybrids. J Appl Pharm Sci., 7(10) 9-15, doi: 10.7324/JAPS.2017.71002.