Synthesis, Molecular Docking Study, Anti-Oxidant and Cytotoxicity Evaluation of New Spiro Six Membered Ring Derivatives of 5-nitro isatin


  • Hala Ayad Mohamed Rasheed Department of Pharmaceutical Chemistry, College of Pharmacy, Mustansiriyah University, Baghdad, Iraq
  • Suaad M. H. Al-Majidi Department of Chemistry, College of Science, University of Baghdad, Baghdad, Iraq



5-nitro isatin, spiro quinozolin-4-one, spiro thiazine-4-one, spiro oxazine-4-one, MCF-7 assay, DPPH assay


Spiro-5-nitro isatino six memberd ring (quinazoline-4-one, thiazine-4-one and oxazine-4-one) respectively were produced by a cycloaddition of 5-nitro isatin Schiff bases [1–5] with anthranilic acid, o-mercapto benzoic acid and salicylic acid. 1HNMR and 13CNMR nuclear magnetic resonance spectroscopies, as well as Fourier-transform infrared spectroscopy, were used to identify the structures of the obtained compounds. These spiro-5-nitro isatin are of interest to us due to their potential antioxidant and anticancer properties. The MTT assay ((3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)) was used to examine in vitro bioactivity testing against breast cancer (MCF-7) cell lines which compound 14 (IC50=79.85, IC50=39) after 24 hours and 48 hours showed good cytotoxicity compared with drug reference tamoxifen. synthesized compounds 6-20 were evaluated for against antioxidant activity by using DPPH assay. In molecular docking, Promising compounds were used to reveal potential cytotoxic activity mechanisms.


E. H. Houssein, M. M. Emam, A. A. Ali and P. N. Suganthan. Deep and machine learning techniques for medical imaging-based breast cancer: A comprehensive review. Expert Systems with Appli. 2021; 167.

A. G. Waks and E. P. Winer. Breast Cancer Treat. A Rev. JAMA. 2019; 321(3): 316.

S. Łukasiewicz, M. Czeczelewski, A. Forma, J. Baj, R. Sitarz and A. Stanisławek. Breast Cancer—Epidemiology, Risk Factors, Classification, Prognostic Markers, and Current Treatment Strategies— An Updated Review. Cancers. 2021; 13 (17): 4287.

M. Bashiri, A. Jarrahpour, S. M. Nabavizadeh et al. Potent antiproliferative active agents: novel bis Schiff bases and bis spiro β-lactams bearing isatin tethered with butylene and phenylene as spacer and DNA/BSA binding behavior as well as studying molecular docking. Med. Chem. Res., 2020; 30: 258–284.

R. E. F. de Paiva, E. G. Vieira and D. R. da Silva et al. Anticancer Compounds Based on Isatin-Derivatives: Strategies to Ameliorate Selectivity and Efficiency. Front. Mol. Biosci., 2021; 7.

G. Chen, H. Su and M. Zhang et al. New bactericide derived from Isatin for treating oilfield reinjection water. Chem. Central J. 2012; 6:90.

G. Kumar, N. Singh and K. Kumar. Recent Advancement of Synthesis of Isatins as a Versatile Pharmacophore: A review. Drug Res. 2021; 71: 115–121.

B. Bhrigua, D. Pathaka and N. Siddiquib et al. Search for Biological Active Isatins: A Short Review. IJPSDR. 2010; 2(4): 229-235.

K. Lahari and R. Sundararajan. Design and synthesis of novel isatin derivatives as potent analgesic, anti-inflammatory and antimicrobial agents. J. Chem. Sci. 2020; 132:94.

İ. S. Doğan, G. G. Bölek and B. Kahveci. Synthesis of Some New Isatin Derivatives and Identification of Their Structures. J. of Natural and Applied Sci. 2019; 23: 67-70.

G. S. Singh and Z. Y. Desta. Isatins as Privileged Molecules in Design and Synthesis of SpiroFused Cyclic Framework. Chem. Rev. 2012;112: 6104−6155.

A. S. Grewal. Isatin Derivatives with Several Biological Activities. IJPR. 2014; 6(10).

R.Paiva, E. Vieira and D. Silva et al. Anticancer Compounds Based on Isatin-Derivatives: Strategies to Ameliorate Selectivity and Efficiency. Frontiers in Molecular Biosciences. 2021; 7.

M. A. Borad, M. N. Bhoi and N. P. Prajapati et al. A Review on Synthesis of Spiro Heterocyclic Compounds from Isatin. Synthetic Communications. 2013.

M. N. Meeran and A. Z. Hussain. Synthesis, Characterization and DPPH Scavenging Assay of Isatin Related Spiroheterocyclic Compounds. Indian J Pharm Sci. 2017; 79(4):641-645.

N. G. Fawazy, S. S. Panda, A. Mostafa and B. M. Kariuki et. al. Development of spiro‑3‑indolin‑2‑one containing compounds of antiproliferative and anti‑SARS‑CoV‑2 properties. scientific reports. 2022; 12:13880.

K. Acosta-Quiroga, C. Rojas-Pena,L and St. Nerio,M. et al. Spirocyclic derivatives as antioxidants: a review. RSC Adv. 2021; 11:21926.

D. J. Connolly and P. J. Guiry. A Facile and Versatile Route to 2-Substituted-4(3H)-Quinazolinones and Quinazolines. LETTER. 2001.

M. Yokoyama. A Novel synthesis of 4H-1,3-thiazine-4-0ne derivatives. Bulletin of the chemical society of Japan. 1971; 44: 1610-1613.

N. Hirose, S. Kuriyama, S. Sohda et. al. Studies on Benzoheterocyclic Derivatives. XIV. Synthesis of Spiro [cycloalkane-1', 2(1H) quinazolin]-4(3H)-ones and the Related Compounds. chem pharm bull. 1972; 20(5) 1005- 1013.

R. Jain and S. Vajpei. reaction of schiff’s bases with mercapto acids. Synthesis of spiro [2h-1,3-benzothiazine-2,3’-[3h] indole]-2’ ,4(1’h,3h)-diones and spir0[3h-indole-3,2’-thiazolidinei-2,4’(1h)-diones) Phosphorus. Sulfur, and Silicon. 1992; 70: 63-66.

T. N. Omar. Synthesis of Schiff Bases of Benzaldehyde and Salicylaldehyde as Anti-inflammatory Agents. Iraqi J.Pharm.Sci., 2007; 16 (2): 17-1.

H. J. Al-Adhami and S. M. H. Al-Majidi. Design, Synthesis, and Antimicrobial and Antioxidant Activities of Some New Dihydrotetrazole, Dihydroquinazolin-4-one, and 1,3-Benzothiazin-4-one Derivatives Based on 6-Amino-1,3-dimethyluracil. Russian J. of Org. Chem. 2021; 57: 448–454.

Hadi, N. A., Mahmood, R. I. and Al-Saffar, A. Z. Evaluation of antioxidant enzyme activity in doxorubicin treated breast cancer patients in Iraq: A molecular and cytotoxic study. Gene Reports. 2021; 24: 101285.

Mahmood, R. I., Abbass, A. Kh., Razali, N., Al-Saffar, A. Z. and Al-Obaidi, J. R. Protein profile of MCF-7 breast cancer cell line treated with lectin delivered by CaCO3NPs revealed changes in molecular chaperones, cytoskeleton, and membrane-associated proteins. International Journal of Biological Macromolecules. 2021; 184: 636-647.

Abdul Rahman, M. N. and Nafiah, M. A. et. al. Antioxidant, Antimicrobial, and Cytotoxic Activities of the Hexane and Dichloromethane extracts of Malaysian Mitragyna spesioca Korth. Leaves. Malaysian Journal of Chemistry. 2022; 24(2): 191-198.

Alvarez J, Shoichet B. Virtual screening in drug discovery. CRC. press; 2005.

K. M. Dnyandev, G. V. Babasaheb, K. V. Chandrashekhar and M. A. Chandrakant et al. A Review on Molecular Docking. IRJPAC. 2021; 22(3): 60-68.

N. S. Pagadala, K. Syed and J. Tuszynski. Software for molecular docking: a review. Biophys Rev. 2017; 9(2):91-102.