Cytotoxic, Docking, and Pharmacokinetic Evaluation of Ethyl Acetate Fraction of Sungkai (Peronema canescens Jack) Leaves Against A549 Lung Cancer Cells
DOI:
https://doi.org/10.31351/vol35iss2pp188-200Keywords:
Peronema canescens Jack, lung cancer, molecular docking, cytotoxicity, pharmacokineticsAbstract
Sungkai (Peronema canescens Jack) is known to have various bioactive compounds as anticancer agents. This study aims to evaluate the anticancer potential of the active compound from the ethyl acetate fraction of sungkai leaves through an experimental and computational approach. The cytotoxic activity of the ethyl acetate fraction was carried out using the MTT assay method on A549 lung cancer cells. The computational study carried out using the molecular docking method and continued with pharmacokinetic analysis of the five main compounds in the ethyl acetate fraction of sungkai leaves. The MTT assay revealed that the ethyl acetate fraction exhibited moderate cytotoxic activity against A549 lung cancer cells, with an IC50 value of 33.41 µg/mL. The LC-MS/MS analysis identified five main compounds in this fraction: isorhamnetin, physcion, pilosin, (3R)-sophorol, and takakin. Molecular docking simulations of these compounds with the EGFR protein (PDB ID: 5UG9) showed bond energies ranging from -7.7 to -8.9 kcal/mol. Physcion has a more negative bond energy value compared to the positive control (quercetin) and four other compounds. The compounds interacted with the protein through hydrogen bonds and van der Waals interactions, predominantly involving amino acid residues ALA A:743, VAL A:726, LEU A:718, LEU A:844, and GLN A:791. Pharmacokinetic analysis revealed that all the compounds met Lipinski's rule of five, indicating good oral absorption and membrane permeation. However, toxicity predictions showed that isorhamnetin, takakin, and pilosin had a high risk of gene mutations, whereas physcion had a moderate risk. Despite potential toxicity concerns, the compounds had drug scores greater than 0, suggesting their potential as drug candidates. Therefore, experimental and computational approaches can be an efficient early screening method to identify potential bioactive compounds as drugs. However, further analysis must be carried out regarding the risk of toxicity and a good dose of the drug.
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References
Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71(3):209-249. doi:10.3322/caac.21660
Zhou J, Xu Y, Liu J, Feng L, Yu J, Chen D. Global burden of lung cancer in 2022 and projections to 2050: Incidence and mortality estimates from GLOBOCAN. Cancer Epidemiol. 2024 ;93 (September) :102693. doi: 10. 1016/j.canep.2024.102693
Sousa AC, Silveira C, Janeiro A, et al. Detection of rare and novel EGFR mutations in NSCLC patients: Implications for treatment-decision. Lung Cancer. 2020;139(October 2019):35-40. doi:10.1016 /j.lungcan. 2019. 10. 030
Prasetiawati R, Suherman M, Permana B, Rahmawati R. Molecular Docking Study of Anthocyanidin Compounds Against Epidermal Growth Factor Receptor (EGFR) as Anti-Lung Cancer. Indones J Pharm Sci Technol. 2021;8(1):8. doi:10.24198/ijpst.v8i1.29872
ArulJothi KN, Kumaran K, Senthil S, et al. Implications of reactive oxygen species in lung cancer and exploiting it for therapeutic interventions. Med Oncol. 2023;40(1):1-19. doi:10.1007/s12032-022-01900-y
Choudhary N, Bawari S, Burcher JT, Sinha D, Tewari D, Bishayee A. Targeting Cell Signaling Pathways in Lung Cancer by Bioactive Phytocompounds. Cancers (Basel). 2023;15(15):1-60. doi:10.3390/cancers15153980
Zhang J, Liu X, Zhang G, et al. To explore the effect of kaempferol on non-small cell lung cancer based on network pharmacology and molecular docking. Front Pharmacol. 2023; 14(July):1-11. doi:10.3389/fphar.2023.1148171
Nurjannah S, Arum D, Lasmana I, Latief M. Anti-inflammatory Prediction of Peronemin Compounds from Sungkai ( Peronema canescens Jack ) and Their Derivatives. 2023;9(2).
Deswita E, Imelda, Suryati. Comparative Analysis of Antioxidant Potential in Hexane and Methanol Fractions of Sungkai Leaves. Hydrog J Kependidikan Kim. 2025;13(June).
Suryati, Irfan Afrinal, Afrizal, Rahmi Vika Ulia. Cytotoxic Potential of Compounds Isolated from Non-Polar Fractions of Sungkai Plant Leaves (Peronema canescens Jack) Against Artemia salina Leach Larvae. J Ris Kim. 2024;15(1):48-60. doi: 10.25077/ jrk.v1 5i1.655
Aulena DN, Yani DF, Mariyamah M, et al. Determination of Flavonoid Content and Anti-Inflammatory Activity Extract and Fraction of Sungkai Leaf (Peronema canescens Jack). J Ilmu Kefarmasian Indones. 2023;21(2):223. doi:10.35814/jifi.v21i2.1437
Ibrahim A, Siswandono, Bambang Prajogo EW. Cytotoxic activity of peronema canescens Jack leaves on human cells: HT-29 and primary adenocarcinoma colon cancer. Pharmacogn J. 2021;13(6):1389-1396. doi:10.5530/PJ.2021.13.176
Vidi GC, Darsono PV, Rahmadani. Toksisitas Ekstrak Daun Sungkai ( Peronema canescens Jack ) Terhadap Larva Artemia salina Leach dengan Metode Brine Shrimp Lethality Test ( BSLT ). J Surya Med. 2024;10(2):129-136.
Amorim AMB, Piochi LF, Gaspar AT, Preto AJ, Rosário-Ferreira N, Moreira IS. Advancing Drug Safety in Drug Development: Bridging Computational Predictions for Enhanced Toxicity Prediction. Chem Res Toxicol. 2024;37(6):827-849. doi:10. 1021 /acs. chemrestox.3c00352
Ahmad I, Ibrahim A. Bioaktivitas Ekstrak Metanol dan Fraksi n-Heksana Daun Sungkai (Peronema canescens JACK) terhadap Larva Udang (Artemia salina Leach). J Sains dan Kesehat. 2015;1(3):114-119. doi:10. 25026 /jsk. v1i3.27
Pertiwi D, Khotimah S, Wardoyo ERP. Uji Aktivitas Antibakteri Fraksi Metanol, Etil Asetat, dan N-Heksana Rimpang Lengkuas Merah (Alpinia purpurata, K. Schum) terhadap Bakteri Propionibacterium acnes. J Protobiont. 2023;12(1):1-8.
Ismed F, Desti WN, Arifa N, Rustini R, Putra DP. TLC-Bioautographic and LC-MS/MS Detection of Antimicrobial Compounds from Four Semipolar Extracts of Cladonia Species . Proc 2nd Int Conf Contemp Sci Clin Pharm 2021 (ICCSCP 2021). 2022;40(Iccscp):49-59. doi:10.2991/ahsr.k.211105.008
Hartanti, Sariyanto I. Hartanti : Perbandingan Aktivitas Penghambatan Sel Kanker Payudara Fraksi Aseton dan Etil Asetat Kulit Buah Coklat (Theobroma cacao) Perbandingan Aktivitas Penghambatan Sel Kanker Payudara Fraksi Aseton dan Etil Asetat Kulit Buah Coklat (Theobroma cacao). J Anal Kesehat. 2024;13(1):28-33.
Fadholly A, Sudjarwo SA, Rantam FA, et al. Uji sitotoksik ekstrak cabai merah keriting (Capsicum annuum) pada sel WiDr secara in vitro. Curr Biomed. 2023;1(2):70-75. doi:10.29244/currbiomed.1.2.70-75
Zulkipli NN, Rahman SA, Taib WRW, et al. The cytotoxicity effect and identification of bioactive compounds of Prismatomeris glabra crude leaf extracts against breast cancer cells. Beni-Suef Univ J Basic Appl Sci. 2024;13(1). doi:10.1186/s43088-024-00490-0
Rajalakshmi R, Lalitha P, Sharma SC, Rajiv A, Chithambharan A, Ponnusamy A. In silico studies: Physicochemical properties, drug score, toxicity predictions and molecular docking of organosulphur compounds against Diabetes mellitus. J Mol Recognit. 2021;34(11). doi:10.1002/jmr.2925
Rajan VK, Ragi C, Muraleedharan K. A Computational Exploration into the Structure, Antioxidant Capacity, Toxicity and Druglike Activity of the Anthocyanidin “Petunidin.” Heliyon. 2019;5(7):e02115.
Ramadhan RG, imelda, Kusuma RT, Phameswari DS. Senyawa Pelargonidin Sebagai Kandidat Obat Menggunakan Metode Dft ( Density Functional Theory ). J Kim Saintek dan Pendidik. 2021;5(2):110-120.
Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading. J Comput Chem. 2010;31(2):455-461. doi:10. 1002/ jcc. 21334. AutoDock
Becke AD. Density-Functional Thermochemistry. III. The Role of Exact Exchange. J Chem Phys. 1993;98(7):5648-5652.
Yusuff OK, Abdul Raheem MAO, Mukadam AA, Sulaimon RO. Kinetics and Mechanism of the Antioxidant Activities of C. olitorius and V. amygdalina by Spectrophotometric and DFT Methods. ACS Omega. 2019;4(9):13671-13680. doi:10.1021/acsomega.9b00851
Tarigan IL, Puspitasari RD, Latief M. Formulation and Characterization of a Microencapsulant of Sungkai Leaves Ethanol Extract (Peronema canescens Jack). Preprints. 2023;1(1):1-17. doi:10.20944 /preprints 202311. 1821.v1
Ulia RV, Suryati, Santoni A. Cytotoxic Potential of Essential Oil Isolated from Semambu (Clibadium surinamese L) Leaves Against T47D Breast and HeLa Cervical Cancer Cells. Molekul. 2023;18(2):289-299. doi:10.20884/1.jm.2023.18.2.7816
Mulia K, Hasan AEZ, Suryani. Total Phenolic, Anticancer and Antioxidant Activity of Ethanol Extract of Piper retrofractum Vahl from Pamekasan and Karang Asem. Curr Biochem. 2016;3(2):80-90. http://biokimia.ipb.ac.id
Stindlova M, Peroutka V, Jencova V, Havlickova K, Lencova S. Application of MTT assay for probing metabolic activity in bacterial biofilm-forming cells on nanofibrous materials. J Microbiol Methods. 2024;224(June):107010. doi:10.1016/j.mimet.2024.107010
Riss TL, Moravec RA, Niles AL, et al. The Assay Guidance manual: Cell Viability Assays. 2016;(Md):1-25.
Suryati S, Santoni A, Ulia R V, Imelda I. Cytotoxic and Molecular Docking Potential of β -Sitosterol Isolated from Lantana camara Leaves against Breast (T47D) and Cervical Cancer (HeLa) Cell Lines. Trop J Nat Prod Res. 2024;8(April):6911-6917.
Budiartina W, Sari WP, Nasution AN, Girsang E. Nanoemulsion of Ethanolic Extract of Sungkai Leaves (Peronema Canescens Jack) for Anti-Acne Therapy. Int J Drug Deliv Technol. 2024;14(3):1604-1610. doi:10.25258/ijddt.14.3.50
Ibrahim A, Siswandono S, Bambang PEW. Anticancer activity of Peronema canescens Jack leaves extracts against human cells: HT-29 and HeLa in vitro. Res J Pharm Technol. 2022;15(10):4739-4745. doi:10. 52711 /0974 -360X.2022.00796
Forli S, Huey R, Pique ME, Sanner MF, & DSG, Olson AJ. Computational Protein–Ligand Docking and Virtual Drug Screening with the AutoDock Suite. Polym J. 2016;48(7):829-834.
Sivaiah G, Raghu MS, Prasad SBB, et al. Synthesis, biological evaluation and molecular docking studies of new pyrimidine derivatives as potent dual EGFR/HDAC inhibitors. J Mol Struct. 2024;1309(January):138223. doi:10. 1016 /j.molstruc.2024.138223
Afriza D, Suriyah W, Ichwan S, Knights joe. Molecular docking analysis between anti-apoptosis EGFR and four coumarins , and four carbazole alkaloids : in silico study. Padjadjaran J Dent. 2024;36(1):117-125. doi :10 . 24198/pjd.vol36no1.52467
Khan Z, Javaid W, Xing L xi. Anthocyanin-Binding Affinity and Non-Covalent Interactions with IIS-Pathway-Related Protein Through Molecular Docking. Published online 2025.
Liu HN, Zhu Y, Chi Y, et al. Synthetic routes and clinical application of Small-Molecule HER2 inhibitors for cancer therapy. Bioorg Chem. 2024;151. doi:10.1016/ j. bioorg .2024.107653
Macabeo APG, Pilapil LAE, Garcia KYM, et al. Phenalenones from a new species of Pseudolophiostoma originating from Thailand. Molecules. 2020;25(4):965-973.
EN-NAHLI F, HAJJI H, OUABANE M, et al. ADMET profiling and molecular docking of pyrazole and pyrazolines derivatives as antimicrobial agents. Arab J Chem. 2023;16(11):105262. doi:10. 1016/j.a rabjc .2023 .105262
Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev. 2001;46(SUPPL.):3-26. doi:10 .1016/ j.addr .2012.09.019
Zeki NM, Mustafa YF. Digital alchemy: Exploring the pharmacokinetic and toxicity profiles of selected coumarin-heterocycle hybrids. Results Chem. 2024 ;10 (August) :101754 . doi:10. 1016/ j. rechem. 2024.101754
Mouli HMC, Harini D, Shaikh N, et al. In silico characterization of indole-substituted densely functionalized pyrrole against breast cancer: Integrating DFT, molecular docking, MD simulations, and ADME analysis. J Mol Struct. 2025;1328(January):141375. doi:10. 1016/ j.molstruc .2025.141375
Elharafi H, Elhamdani N, Hachim ME, et al. In silico exploration of bioavailability, druggability, toxicity alerts and biological activity of a large series of fatty acids. Comput Toxicol. 2021;17(November 2020):100153. doi: 10. 1016/j.comtox.2021.100153
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