Chemical Profiling and In Silico Study of Russelia equisetifomis Major Phytochemicals as Potential Dual-Site Acetylcholinesterase Inhibitors in Alzheimer’s Disease
DOI:
10.29303/jpm.v21i5.12030Published:
2026-09-30Downloads
Abstract
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline and memory impairment, associated with cholinergic dysfunction and β-amyloid aggregation. Acetylcholinesterase (AChE) plays a dual role in AD progression by hydrolysing acetylcholine (ACh) at the catalytic active site (CAS) and promoting β-amyloid aggregation at the peripheral anionic site (PAS), making dual-site inhibition a promising therapeutic strategy. This study aimed to evaluate the potential of major phytochemicals from the stem of Russelia equisetiformis as dual-site AChE inhibitors using an in silico approach. Chemical profiling of the ethanolic stem extract was performed using Liquid Chromatography-Mass Spectrophotometry (LC-MS), and compounds with relative concentrations above 2% were selected for further analysis. Molecular docking was conducted using Autodock Vina implemented in PyRx against AChE (PDB ID: 4M0E), while ligand interactions were visualised using BIOVIA Discovery Studio Visualizer. Druglikeness was evaluated using SwissADME based on Lipinski’s Rule of Five, and Molecular Initiating Event (MIE) prediction was performed using ProTox 3.0. Docking validation produced an RMSD value of 1.672 Å, indicating acceptable reliability. Among the identified compounds, tilianin exhibited a binding affinity of -8.5 kcal/mol and interacted with key PAS residues (Tyr72, Trp286, and Tyr341) as well as residues extending toward the CAS, comparable to that of donepezil (-7.4 kcal/mol). Tilianin also fulfilled all Lipinski criteria and was predicted to be active toward AChE with a probability score of 0.65. These findings suggest that tilianin is a promising dual-site AChE inhibitor candidate that may simultaneously improve cholinergic neurotransmission and reduce β-amyloid (Aβ) aggregation. Nevertheless, further in vitro and in vivo validation studies are required to confirm its therapeutic potential against AD.
Keywords:
Alzheimer's Disease Acetylcholinesterase Dual-Site Molecular Docking Russelia equisetiformisReferences
[1] H. Karagecili, E. İzol, E. Kirecci, and İ. Gulcin, “Determination of Antioxidant, Anti-Alzheimer, Antidiabetic, Antiglaucoma and Antimicrobial Effects of Zivzik Pomegranate (Punica granatum)—A Chemical Profiling by LC-MS/MS,” Life 2023, Vol. 13, Page 735, vol. 13, no. 3, p. 735, Mar. 2023, doi: 10.3390/LIFE13030735.
[2] WHO, “Dementia.” Accessed: Apr. 29, 2026. [Online]. Available: https://www.who.int/news-room/fact-sheets/detail/dementia
[3] WHO, “Dementia.” Accessed: May 01, 2026. [Online]. Available: https://www.who.int/news-room/fact-sheets/detail/dementia
[4] R. Tenchov, J. M. Sasso, and Q. A. Zhou, “Alzheimer’s Disease: Exploring the Landscape of Cognitive Decline,” ACS Chem. Neurosci., vol. 15, no. 21, p. 3800, Nov. 2024, doi: 10.1021/ACSCHEMNEURO.4C00339.
[5] Z. Breijyeh, R. Karaman, D. Muñoz-Torrero, and R. Dembinski, “Comprehensive Review on Alzheimer’s Disease: Causes and Treatment,” Molecules 2020, Vol. 25, Page 5789, vol. 25, no. 24, p. 5789, Dec. 2020, doi: 10.3390/MOLECULES25245789.
[6] J. Zhang, Y. Zhang, J. Wang, Y. Xia, J. Zhang, and L. Chen, “Recent advances in Alzheimer’s disease: mechanisms, clinical trials and new drug development strategies,” Signal Transduction and Targeted Therapy 2024 9:1, vol. 9, no. 1, pp. 211-, Aug. 2024, doi: 10.1038/s41392-024-01911-3.
[7] Y. Pourshojaei, A. Abiri, K. Eskandari, Z. Haghighijoo, N. Edraki, and A. Asadipour, “Phenoxyethyl Piperidine/Morpholine Derivatives as PAS and CAS Inhibitors of Cholinesterases: Insights for Future Drug Design,” Scientific Reports 2019 9:1, vol. 9, no. 1, pp. 19855-, Dec. 2019, doi: 10.1038/s41598-019-56463-2.
[8] P. Muñoz-Ruiz et al., “Design, Synthesis, and Biological Evaluation of Dual Binding Site Acetylcholinesterase Inhibitors: New Disease-Modifying Agents for Alzheimer’s Disease,” J. Med. Chem., vol. 48, no. 23, pp. 7223–7233, Nov. 2005, doi: 10.1021/JM0503289.
[9] K. Adebambo and O. C. Ojoh, “In Silico Investigation of Novel Compounds as Inhibitors of Acetylcholinesterase Enzyme for the Treatment of Alzheimer’s Diseases,” Int. J. Alzheimers Dis., vol. 2024, no. 1, p. 2988685, Jan. 2024, doi: 10.1155/2024/2988685.
[10] A. A. Bagalagel et al., “The Protective and Therapeutic Anti-Alzheimer Potential of Olea europaea L. cv. Picual: An In Silico and In Vivo Study,” Metabolites 2022, Vol. 12, Page 1178, vol. 12, no. 12, p. 1178, Nov. 2022, doi: 10.3390/METABO12121178.
[11] M. A. Fakhirah, P. Husni, J. Raya, B. Sumedang, and K. 21 Jatinangor, “Review Artikel: Potensi Tanaman Herbal sebagai Agen Terapi Penyakit Alzheimer,” Farmaka, vol. 22, no. 2, Jul. 2024, doi: 10.24198/FARMAKA.V22I2.54888.
[12] H. Khan, Marya, S. Amin, M. A. Kamal, and S. Patel, “Flavonoids as acetylcholinesterase inhibitors: Current therapeutic standing and future prospects,” Biomed. Pharmacother., vol. 101, pp. 860–870, May 2018, doi: 10.1016/J.BIOPHA.2018.03.007.
[13] U. Khan et al., “Natural alkaloids with therapeutic potential against Alzheimer’s disease through cholinesterase inhibition,” Eur. J. Med. Chem., vol. 302, p. 118371, Jan. 2026, doi: 10.1016/J.EJMECH.2025.118371.
[14] L. S. Min et al., “Plant Terpenoids as the Promising Source of Cholinesterase Inhibitors for Anti-AD Therapy,” Biology 2022, Vol. 11, Page 307, vol. 11, no. 2, p. 307, Feb. 2022, doi: 10.3390/BIOLOGY11020307.
[15] J. B. Johnson, P. Neupane, S. P. Bhattarai, T. Trotter, and M. Naiker, “Phenolic profiles and acetylcholinesterase inhibitory activity in Australian adzuki bean,” Food Chemistry Advances, vol. 3, p. 100361, Dec. 2023, doi: 10.1016/J.FOCHA.2023.100361.
[16] M. A. Fouad, E. M. Ahmed, S. Y. Desoukey, and M. S. Kamel, “Chemical and Biological Studies of Russelia equisetiformis (Sch.&Cham.) Aerial Parts,” J App Pharm, vol. 8, no. 2, pp. 23–33, 2016, doi: 10.21065/19204159.
[17] Y. Yadav and D. Chumbhale, “DMBA induced mammary tumor study of Phyto-fabricated sliver nanoparticles from Russelia equisetiformis flower extract in rat,” Int. J. Health Sci. (Qassim)., pp. 4348–4367, Jul. 2022, doi: 10.53730/ijhs.v6ns6.10278.
[18] Muhammad Riaz, “Antioxidant, antimicrobial and cytotoxicity studies of Russelia equisetiformis,” Afr. J. Microbiol. Res., vol. 6, no. 27, Jul. 2012, doi: 10.5897/ajmr12.800.
[19] O. H. Olusegun, “Hepatoprotective Potential of Russelia equisetiformis Plant Extract on Drug-Induced Hepatotoxicity in Experimental Models,” Journal of Medical science and clinical research, vol. 5, no. 7, Jul. 2017, doi: 10.18535/jmscr/v5i7.85.
[20] N. Cichon et al., “Mechanistic and Therapeutic Insights into Flavonoid-Based Inhibition of Acetylcholinesterase: Implications for Neurodegenerative Diseases,” Nutrients 2025, Vol. 17, Page 78, vol. 17, no. 1, p. 78, Dec. 2024, doi: 10.3390/NU17010078.
[21] P. Zhou and F. Hua, “Exploration of Acetylcholinesterase Inhibitors from Flavonoids and Flavonoid Glycosides,” Neurochemical Journal 2020 14:3, vol. 14, no. 3, pp. 251–256, Sep. 2020, doi: 10.1134/S1819712420030137.
[22] D. D. Soejarto, E. M. Addo, and A. D. Kinghorn, “The continuing need for taxonomic input in phytochemical research,” J. Ethnopharmacol., vol. 354, p. 120474, Jan. 2026, doi: 10.1016/J.JEP.2025.120474.
[23] B. Durhan, E. Yalçın, K. Çavuşoğlu, and A. Acar, “Molecular docking assisted biological functions and phytochemical screening of Amaranthus lividus L. extract,” Scientific Reports 2022 12:1, vol. 12, no. 1, pp. 4308-, Mar. 2022, doi: 10.1038/s41598-022-08421-8.
[24] A. M. Sururi, M. Raihan, E. R. Aisa, F. N. Safitri, I. C. Constaty, and Tukiran, “Anti-Inflammatory Activity of Stem Bark Dichloromethane Fraction Syzygium samarangense Extract as COX-2 Inhibitor: A Bioinformatics Approach,” Jurnal Kimia Riset, vol. 7, no. 2, pp. 94–100, Dec. 2022, doi: 10.20473/JKR.V7I2.39662.
[25] N. Stern et al., “Dual Inhibitors of AChE and BACE-1 for Reducing Aβ in Alzheimer’s Disease: From In Silico to In Vivo,” Int. J. Mol. Sci., vol. 23, no. 21, p. 13098, Nov. 2022, doi: 10.3390/IJMS232113098/S1.
[26] B. Saha, A. Das, K. Jangid, A. Kumar, V. Kumar, and V. Jaitak, “Identification of coumarin derivatives targeting acetylcholinesterase for Alzheimer’s disease by field-based 3D-QSAR, pharmacophore model-based virtual screening, molecular docking, MM/GBSA, ADME and MD Simulation study,” Curr. Res. Struct. Biol., vol. 7, p. 100124, Jan. 2024, doi: 10.1016/J.CRSTBI.2024.100124.
[27] S. Johri et al., “Inspection of in-house designed novel thiochromone amino-acid conjugate derivatives as Lm-NMT inhibitor – An in-silico analysis,” J. Mol. Graph. Model., vol. 119, Mar. 2023, doi: 10.1016/j.jmgm.2022.108397.
[28] C. A. Lipinski, F. Lombardo, B. W. Dominy, and P. J. Feeney, “Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings,” Adv. Drug Deliv. Rev., vol. 46, no. 1–3, pp. 3–26, Mar. 2001, doi: 10.1016/S0169-409X(00)00129-0.
[29] D. Gadaleta, N. Spînu, A. Roncaglioni, M. T. D. Cronin, and E. Benfenati, “Prediction of the Neurotoxic Potential of Chemicals Based on Modelling of Molecular Initiating Events Upstream of the Adverse Outcome Pathways of (Developmental) Neurotoxicity,” Int. J. Mol. Sci., vol. 23, no. 6, p. 3053, Mar. 2022, doi: 10.3390/IJMS23063053/S1.
[30] I. Y. Wardhani, A. H. Ramadani, F. Widyaningrum, and V. Famelia, “The Effect of Extraction Method on Total Flavonoid Content of Ageratum conyzoides Ethanol Extract,” Journal Of Biology Education, vol. 6, no. 2, pp. 134–145, Dec. 2023, doi: 10.21043/JOBE.V6I2.23398.
[31] B. Santoso et al., “Procurement and Operation Technical For Meniran (Phyllanthus Niruri) Extraction Equipment,” Mekanika: Majalah Ilmiah Mekanika, vol. 20, no. 1, pp. 34–43, Mar. 2021, doi: 10.20961/mekanika.v20i1.45487.
[32] D. G. Aguila-Muñoz et al., “Synthesis and Molecular Docking Studies of Alkoxy- and Imidazole-Substituted Xanthones as α-Amylase and α-Glucosidase Inhibitors,” Molecules, vol. 28, no. 10, p. 4180, May 2023, doi: 10.3390/MOLECULES28104180/S1.
[33] P. C. Agu et al., “Molecular docking as a tool for the discovery of molecular targets of nutraceuticals in diseases management,” Sci. Rep., vol. 13, no. 1, p. 13398, Dec. 2023, doi: 10.1038/S41598-023-40160-2.
[34] M. Bartolini, C. Bertucci, V. Cavrini, and V. Andrisano, “β-Amyloid aggregation induced by human acetylcholinesterase: Inhibition studies,” Biochem. Pharmacol., vol. 65, no. 3, pp. 407–416, Feb. 2003, doi: 10.1016/S0006-2952(02)01514-9.
[35] F. J. Carvajal, N. C. Inestrosa, J. Saez-Valero, and U. Miguel Hernandez, “Interactions of AChE with Aβ Aggregates in Alzheimer’s Brain: Therapeutic Relevance of IDN 5706,” Front. Mol. Neurosci., vol. 4, p. 12633, Sep. 2011, doi: 10.3389/FNMOL.2011.00019.
[36] A. Kračmarová, L. Drtinová, and M. Pohanka, “Possibility of Acetylcholinesterase Overexpression in Alzheimer Disease Patients after Therapy with Acetylcholinesterase Inhibitors,” Acta Medica (Hradec Kralove), vol. 58, no. 2, pp. 37–42, 2015, doi: 10.14712/18059694.2015.91.
[37] T. L. Rosenberry et al., “Comparison of the Binding of Reversible Inhibitors to Human Butyrylcholinesterase and Acetylcholinesterase: A Crystallographic, Kinetic and Calorimetric Study,” Molecules, vol. 22, no. 12, Dec. 2017, doi: 10.3390/MOLECULES22122098.
[38] R. M. Lane, S. G. Potkin, and A. Enz, “Targeting acetylcholinesterase and butyrylcholinesterase in dementia,” Int. J. Neuropsychopharmacol., vol. 9, no. 1, pp. 101–124, Feb. 2006, doi: 10.1017/S1461145705005833.
[39] M. A. Al-Hilo, “Computational Investigation of Natural Flavonoids as Inhibitors of Human Acetylcholinesterase (AChE) in Alzheimer’s Disease,” Cognizance Journal of Multidisciplinary Studies, vol. 5, no. 10, pp. 1–15, Oct. 2025, doi: 10.47760/cognizance.2025.v05i10.001.
[40] J. Wu, X. Kou, H. Ju, H. Zhang, A. Yang, and R. Shen, “Design, synthesis and biological evaluation of naringenin carbamate derivatives as potential multifunctional agents for the treatment of Alzheimer’s disease,” Bioorg. Med. Chem. Lett., vol. 49, p. 128316, Oct. 2021, doi: 10.1016/J.BMCL.2021.128316.
[41] R. Roskoski, “Rule of five violations among the FDA-approved small molecule protein kinase inhibitors,” Pharmacol. Res., vol. 191, p. 106774, May 2023, doi: 10.1016/J.PHRS.2023.106774.
[42] Y. Y. Liu, X. Y. Feng, W. Q. Jia, Z. Jing, W. R. Xu, and X. C. Cheng, “Identification of novel PI3Kδ inhibitors by docking, ADMET prediction and molecular dynamics simulations,” Comput. Biol. Chem., vol. 78, pp. 190–204, Feb. 2019, doi: 10.1016/j.compbiolchem.2018.12.002.
[43] D. N. Olennikov, О. Д. Н., N. I. Kashchenko, and К. Н. И., “New Acacetin Glycosides and Other Phenolics from Agastache foeniculum and Their Influence on Monoamine Oxidase A and B,” Appl. Biochem. Microbiol., vol. 60, no. 6, pp. 645–656, Nov. 2024, doi: 10.31857/S0555109924060084.
[44] F. Fusi, A. Trezza, M. Tramaglino, G. Sgaragli, S. Saponara, and O. Spiga, “The beneficial health effects of flavonoids on the cardiovascular system: Focus on K+ channels,” Pharmacol. Res., vol. 152, Feb. 2020, doi: 10.1016/J.PHRS.2019.104625.
[45] M. A. Abdel-Rasol, R. A. R. Abbas, and W. M. El-Sayed, “Tilianin: pharmacological potential, mechanisms of action, and future perspectives in traditional and modern medicine,” Chin. Med., vol. 20, no. 1, Dec. 2025, doi: 10.1186/S13020-025-01245-4.
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