Author(s):
Suganya Ilango, Priyanka Jayachandran, Ramalingam Nirmaladevi
Email(s):
suganyai@srmist.edu.in
DOI:
10.52711/0974-360X.2026.00334 
Address:
Suganya Ilango1*, Priyanka Jayachandran2, Ramalingam Nirmaladevi3
1Department of Biochemistry, Faculty of Science and Humanities, SRM Institute of Science and Technology, Kattankulathur Campus, Chengalpattu-603203, Tamil Nadu, India.
2Department of Biochemistry and Biotechnology, Avinashilingam Institute for Home Science and Higher Education for Women, Coimbatore 641108, Tamil Nadu, India.
3Department of Biochemistry, Biotechnology and Bioinformatics, Avinashilingam Institute for Home Science and Higher Education for Women, Coimbatore-641043, Tamil Nadu, India.
*Corresponding Author
Published In:
Volume - 19,
Issue - 5,
Year - 2026
ABSTRACT:
Annona muricata (soursop) leaves are rich in bioactive compounds with known anticancer properties. Flavonoids and acetogenins, key constituents of these leaves, have shown potential antileukemic effects individually, but their combined efficacy and selectivity remain underexplored. The primary aim of the research is to examine the synergistic anticancer effects of flavonoid and acetogenin enriched fractions from A. muricata leaves on leukemic (Molt-3) cells and assess their cytotoxicity on normal peripheral blood lymphocytes (PBL). Ethanolic extracts of A. muricata leaves were subjected to bioassay-guided fractionation and chromatographic separation to isolate flavonoid and acetogenin enriched fractions. GC-MS was used to identify phytoconstituents. Cytotoxicity was assessed using MTT assay, and synergy was analysed using Combenefit software based on Loewe, Bliss, and Highest Single Agent models. Apoptosis and cell cycle changes were evaluated using flow cytometry. GC-MS analysis revealed the detection of flavonoids such as myricetin and luteolin glucosides, and acetogenins like muricatacin and a-muurolene. The combination of flavonoid and acetogenin fractions exhibited enhanced cytotoxicity in Molt-3 cells (IC50: 40.21 µg/ml) compared to individual treatments. Synergy was confirmed across all models, particularly at 60 µg/ml of flavonoids and 80–100 µg/ml of acetogenins. Flow cytometry showed increased G0/G1 arrest and apoptosis in Molt-3 cells, while PBLs remained largely unaffected, indicating selectivity. Flavonoid and acetogenin fractions from A. muricata exert synergistic, selective cytotoxic effects on leukemic cells while sparing normal cells, suggesting their potential as a natural, targeted therapeutic approach for leukemia.
Cite this article:
Suganya Ilango, Priyanka Jayachandran, Ramalingam Nirmaladevi. Synergistic Anticancer Effects of Flavonoid and Acetogenin Fractions from Annona muricata Leaves in Leukemic and Peripheral Blood Cells. Research Journal Pharmacy and Technology. 2026;19(5):2329-5. doi: 10.52711/0974-360X.2026.00334
Cite(Electronic):
Suganya Ilango, Priyanka Jayachandran, Ramalingam Nirmaladevi. Synergistic Anticancer Effects of Flavonoid and Acetogenin Fractions from Annona muricata Leaves in Leukemic and Peripheral Blood Cells. Research Journal Pharmacy and Technology. 2026;19(5):2329-5. doi: 10.52711/0974-360X.2026.00334 Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2026-19-5-55
REFERENCES:
1. Suhail P et al. A review on the in vitro anticancer potentials of acetogenins from Annona muricata Linn.: A potential inducer of Bax-Bak and caspase-3 related pathways. Indian J Pharm Educ Res. 2024; 58(3s): s693–703. https://doi.org/10.5530/ijper.58.3s.73.
2. Adido HEF et al. In silico studies on cytotoxicity and antitumoral activity of acetogenins from Annona muricata L. Front Chem. 2023; 11: 1316779. https://doi.org/10.3389/fchem.2023.1316779.
3. Abdallah RH et al. LCMS/MS Phytochemical Profiling, Molecular, Pathological, and Immune-Histochemical Studies on the Anticancer Properties of Annona muricata. Molecules. 2023; 28: 5744. https://doi.org/10.3390/molecules28165744.
4. Okoro Ebere Kelechi, Amaku Friday James, Okoro Innocent Ajah. Characterization of Annona muricata stem methanol extract by gas chromatography–mass spectrometry analysis. Res. J. Pharmacognosy and Phytochem. 2016; 8(3): 165-171.
5. Widad M.K. Al-ani, Sarah S. Almahdawy. GC/MS Analysis of the Peels of Annona muricata L. Research Journal of Pharmacy and Technology. 2022; 15(7): 3137-0.
6. Pieme CA et al. Antiproliferative activity of Annona muricata extract on human cancer cell lines. BMC Complement Altern Med. 2014; 14: 516. https://doi.org/10.1186/1472-6882-14-516.
7. Moghadamtousi SZ et al. Annona muricata (Annonaceae): A review of its traditional uses, isolated acetogenins and biological activities. Int J Mol Sci. 2015; 16(7): 15625–58. https://doi.org/10.3390/ijms160715625.
8. Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. Sci World J. 2013; 2013: 162750. https://doi.org/10.1155/2013/162750.
9. Imran M et al. Flavonoids as potent antioxidants: A review. J Food Biochem. 2019; 43(3): e12774. https://doi.org/10.1111/jfbc.12774.
10. Suganya Ilango, Ramalingam Nirmaladevi, “Bioactive Leaf Fractions of Annona muricata Exhibit Cytotoxicity Against MOLT-3 Leukemic Cells,” Indian Journal of Nutrition and Dietetics, 2025.
11. Pattilthodika Suhail, Velappan Venkatachalam Venkatachalam, Sanal Dev, Radhakrishnan Murali, Arvind Babu Lalpet Renganathan. Quinic acid from Annona muricata Fruit Inhibits Cyclophosphamide Metabolism through Modulation of Cytochrome P450: Implications for Herbal-Drug Interaction. Research Journal of Pharmacy and Technology. 2025; 18(6): 2664-9.
12. S.B. Santhosh, M.J.N. Chandrasekar, Kaviarasan. L, P. Deepak, T. Silambarasan, B. Gayathri, D. Natarajan. Chemical Composition, Antibacterial, Anti-oxidant and Cytotoxic properties of Green Synthesized Silver Nanoparticles from Annona muricata L. (Annonaceae).Research J. Pharm. and Tech. 2020; 13(1): 33-39.
13. Arvindganth. R, Anupriya K.V., Kathiravan. G. Enhancement of Anticancer Drug Annona muricata Against HT-29 Cell Line using Silver Nano Particles. Research J. Pharm. and Tech. 2017; 10(2): 529-532.
14. Merin Babu, Subin Varghese Thomas, Sruthi T P, Jipnomon Joseph. Evaluation of Cytotoxic Activity of Annona muricata Fruits and Leaves. Research J. Pharm. and Tech. 2019; 12(8): 3802-3806.
15. Merin Babu et al. Evaluation of Cytotoxic Activity of Annona muricata Fruits and Leaves. Research J. Pharm. and Tech. 2019; 12(8): 3802-3806. doi: 10.5958/0974-360X.2019.00651.6.
16. Banerjee S et al. Apigenin downregulates CDKs and cyclins and induces G0/G1 arrest in prostate cancer cells. J Cell Biochem. 2007; 102(3): 661–72.
17. Baskar AA et al. Chemopreventive effect of Annona muricata in hepatocellular carcinoma. Nutr Cancer. 2016; 68(2): 254–60.
18. Bermejo A et al. Acetogenins from Annonaceae: Recent progress in isolation, synthesis and mechanisms of action. Nat Prod Rep. 2005; 22(2): 269–303. https://doi.org/10.1039/B500186M.
19. D’Angelo S et al. Antiproliferative effects of Annona muricata in prostate cancer. J Med Food. 2020; 23(3): 340–9.
20. Fofaria NM, Srivastava SK. Nanoformulations of flavonoids for cancer therapy. Cancer Lett. 2016; 370(2): 185–91.
21. Fulda S. Apoptosis pathways in cancer and cancer therapy. Cancer Immunol Immunother. 2009; 58(10): 1533–40.
22. Berenbaum MC. What is synergy? Pharmacol Rev. 1989; 41(2): 93–141. https://doi.org/10.1016/S0031-6997(25)00026-2.
23. Chauhan A et al. In vivo evaluation of Annona muricata in skin carcinoma. Pharm Biol. 2020; 58(1): 567–75.
24. Cheng Y et al. Synergistic drug combinations in cancer therapy. Acta Pharm Sin B. 2019; 9(1): 4–27.
25. Degli Esposti M. Mitochondrial complex I inhibitors and cell death: Relevance for neurodegenerative and cancer therapy. FEBS Lett. 1998; 426(1): 1–6.
26. Aranha MM et al. Apoptosis-inducing factor mediates apoptosis in response to poly(ADP-ribose) polymerase-1 hyperactivation. Biochem J. 2007; 385(3): 849–57. https://doi.org/10.1042/BJ20041542.
27. Bailly C. Annona muricata: A mini-review of its anticancer activities. Curr Drug Targets. 2020; 21(7): 700–10. https://doi.org/10.2174/1389450120666190613113517.
28. Li Y et al. Dietary flavonoids as cancer chemopreventive agents. J Nutr Biochem. 2007; 18(7): 415–32.
29. Merin Babu et al. Evaluation of Cytotoxic Activity of Annona muricata Fruits and Leaves. Research J. Pharm. and Tech. 2019; 12(8): 3802-3806. doi: 10.5958/0974-360X.2019.00651.6
30. Loewe S. The problem of synergism and antagonism of combined drugs. Arzneimittelforschung. 1953; 3(6): 285–90.
31. Luo Y et al. Improving bioavailability of flavonoids via nanocarriers. Adv Drug Deliv Rev. 2020; 159: 66–81.
32. Vijay R Salunkhe, Satish B Bhise. Synergistic Effect of Natural Sweetener on Antidiabetic Potential of Madhujeevan churna. Research J. Pharmacognosy and Phytochemistry. 2009; 1(3): 204-208.
33. Niran A. Al-Ogaili, Safa Osama, Doha Jazme, Sara Saad. In Vitro Antibacterial investigation and Synergistic Effect of Ficus carica and Olea europaea Aqueous Extracts. Research J. Pharm. and Tech. 2020; 13(3): 1198-1203.
34. Hadeel Esam Hassan, Sundus Hameed Ahmed. Synergistic effect of Moringa Leaves and Antifungal on Candida albicans. Research Journal of Pharmacy and Technology. 2023; 16(3): 1369-4.
35. Pavithra R C, Arun A, Kanimozhi S. Screening of Phytochemical and Quantitative Lipase Inhibition of Honey and Cinnamon Paste: A Synergistic Effect. Research Journal of Pharmacy and Technology. 2023; 16(8): 3709-3.
36. Gautam MK et al. Nanoparticles for cancer therapy: Enhancing drug delivery. Curr Drug Metab. 2019; 20(3): 187–96.
37. Gavamukulya Y et al. Cytotoxicity of Annona muricata leaf extracts. Afr Health Sci. 2014; 14(4): 850–61.
38. Benavides A et al. Synergistic apoptotic effects of flavonoid combinations in leukemic cell lines. Toxicol In Vitro. 2020; 67: 104913. https://doi.org/10.1016/j.tiv.2020.104913.
39. Ko H et al. Graviola extract inhibits prostate cancer through downregulation of EGFR and induction of apoptosis. Oncol Lett. 2011; 2(5): 821–8.
40. Dandawate PR et al. Targeting cancer stem cells and signaling pathways by phytochemicals: Novel approach for breast cancer therapy. Semin Cancer Biol. 2016; 40–41: 192–208. https://doi.org/10.1016/j.semcancer.2016.02.003.
41. Li X et al. Systems pharmacology in traditional Chinese medicine: Strategies and applications. Pharmacol Ther. 2015; 151: 28–42.
42. McLaughlin JL. Paw paw and cancer: Annonaceous acetogenins from discovery to commercial products. J Nat Prod. 2008; 71(8): 1311–21. https://doi.org/10.1021/np800191t.
43. Mital A et al. Annona muricata exhibits selective cytotoxicity in colon and breast cancer cells. J Ethnopharmacol. 2020; 252: 112603.
44. Sakshi D. Patil et al. A Short Review on Anticancer Fruit Annona muricata. Research Journal of Pharmacognosy and Phytochemistry. 2024; 16(4): 270-7. doi: 10.52711/0975-4385.2024.00050.
45. Arvindganth et al. Enhancement of Anticancer Drug Annona muricata Against HT-29 Cell Line using Silver Nano Particles. Research J. Pharm. and Tech. 2017; 10(2): 529-532. doi: 10.5958/0974-360X.2017.00105.6.
46. Pal HC et al. Apigenin induces cell death through mitochondrial pathway in melanoma. Cell Death Dis. 2013; 4:e620.
47. Pathak N et al. Annona muricata mediated apoptosis in murine models. Phytother Res. 2016; 30(5): 795–801.
48. Pathak S et al. Cell cycle arrest and apoptosis by quercetin and luteolin. Cancer Lett. 2007; 254(2): 192–206.
49. Pelicano H et al. ROS stress in cancer therapy. J Cell Biochem. 2004; 97(2): 233–44. https://doi.org/10.1016/j.drup.2004.01.004.
50. Hseu YC et al. Apigenin induces apoptosis through mitochondrial dysfunction in human leukemia cells. Environ Toxicol. 2014; 29(10): 1171–84. https://doi.org/10.1002/tox.21853.
51. Rady I et al. Graviola extract reduces breast cancer cell viability. Nutr Cancer. 2018; 70(1): 121–9. https://doi.org/10.1155/2018/1826170.
52. Ramos S. Effects of flavonoids on cell cycle and apoptosis in cancer cells. Curr Nutr Food Sci. 2008;4(1):20–31.
53. Schumacker PT. Reactive oxygen species in cancer cells: Live by the sword, die by the sword. Cancer Cell. 2006; 10(3): 175–6. https://doi.org/10.1016/j.ccr.2006.08.015.
54. Seelinger G et al. Anti-cancer effects of flavonoids: Molecular mechanisms. Curr Med Chem. 2008; 15(2): 176–86.
55. Iqbal J, et al. Plant-derived anticancer agents: A green anticancer approach. Asian Pac J Trop Biomed. 2017; 7(12): 1129–50. https://doi.org/10.1016/j.apjtb.2017.10.008.
56. Tang J et al. Drug combination prediction using a deep learning model. BMC Bioinformatics. 2020; 21: 487.
57. Torres MP et al. The effect of Annona muricata extract on pancreatic cancer cells. J Cancer Sci Ther. 2012; 4(8): 264–70.
58. Trachootham D et al. Targeting cancer cells by ROS-mediated mechanisms: A radical therapeutic approach? Nat Rev Drug Discov. 2009; 8(7): 579–91. https://doi.org/10.1038/nrd2803.
59. Verma AK et al. G1 phase cell cycle arrest and apoptosis in colon cancer cells by luteolin. Cancer Lett. 2009; 285(2): 147–57.
60. Wagner H. Synergy research: How to optimize synergistic effects in phytotherapy. J Ethnopharmacol. 2011; 134(1): 1–6.
61. Williamson EM. Synergy and other interactions in phytomedicines. Phytomedicine. 2001; 8(5): 401–9. https://doi.org/10.1078/0944-7113-00060.
62. Yadav B, et al. Searching for drug synergy in complex dose-response landscapes using Combenefit. Bioinformatics. 2015; 31(18): 2891–3.
63. Yuan SS, et al. Annonacin, a natural mitochondrial complex I inhibitor, induces apoptosis in cancer cells. Cancer Lett. 2003; 190(1): 63–71.
64. Zhang L, et al. Nanotechnology in cancer therapy. Nat Rev Cancer. 2008; 8(9): 722–32.
65. Zhang Q et al. Synergistic antitumor effects of apigenin and paclitaxel. J Cell Mol Med. 2019; 23(1): 510–20.
66. Zhao L et al. Caspase activation pathway in cancer cell apoptosis. J Cell Biochem. 2012; 113(8): 2645–52. https://doi.org/10.1002/jcp.23006.
67. Zhou Y et al. Role of mitochondria in cancer chemotherapy resistance. Cell Death Dis. 2020; 11(4): 1–15.
68. Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. J Nat Prod. 2020; 83(3): 770–803. https://doi.org/10.1021/acs.jnatprod.9b01285.
69. Okagu OD, Udenigwe CC. Plant bioactives as modulators of oxidative stress in cancer therapy. Curr Res Food Sci. 2022; 5: 2140–54. https://doi.org/10.1016/j.crfs.2022.11.004.
70. Park C et al. Induction of apoptosis and cell cycle arrest by isoferulic acid in human leukemic cells. Int J Mol Sci. 2019; 20(5): 1238. https://doi.org/10.3390/ijms20051238.
71. Tundis R et al. Natural products as antitumor agents: Current status and future perspectives. Curr Med Chem. 2008; 15(34): 3477–95. https://doi.org/10.2174/092986708786848815.