Alzheimer’s and Parkinson’s diseases are the most common neurodegenerative disorders of the central nervous system, characterized by progressive neuronal death and neurological dysfunction. There is currently no treatment that effectively slows the disease progression, and the synthetic drugs proposed to alleviate the symptoms often cause side effects. Many studies are now focused on neuroprotective properties exhibited by natural agents, such as polyphenols, whose action on different cell signaling pathways is well known. In this article, we analyzed the composition and the properties of polyphenolic extracts from forage plants, such as Lotus ornithopodioides, Hedysarum coronarium, Medicago sativa, and Cichorium intybus. The liquid chromatography-mass spectrometry/mass spectrometry analysis on the extracts allowed the identification of total 24 phenolic acids and 25 flavonoids. The effects of the extracts on key enzymes of cholinergic neurotransmission, such as acetyl cholinesterase and butyryl cholinesterase, were examined, together with an investigation on the aggregation and disaggregation of amyloid fibrils. The polyphenols acted as inhibitors of the considered enzymes and interfered with the amyloidogenesis process, with differences depending on the specific extracts. The inhibition constants towards cholinesterases ranged in the 60–240 μM interval; the extracts showed different inhibition mechanisms, from competitive to non-competitive. Differences also emerged in the amyloidogenesis process, with IC50 values comprised in a large interval. Finally, the extract from M. sativa significantly reduced the cell viability of the human neuroblastoma cell line SH-SY5Y. These results suggest that polyphenols extracted from these plants may behave as multitargeting agents against key factors of Alzheimer’s and Parkinson’s diseases. Therefore, they can be considered as promising candidates for the prevention and management of symptoms of these neurodegenerative disorders in combination with pharmacological therapies.
D’Errico A, Reveglia P, Nasso R, Maugeri A, Masullo M, Arcone R, Lecce L, De Vendittis E, Rullo R. Effects of polyphenolic extracts from Mediterranean forage crops on cholinesterases and amyloid aggregation relevant to neurodegenerative diseases. J Appl Pharm Sci. 2025. Article in Press. http://doi.org/10.7324/JAPS.2026.251960
1. Zanetti O, Solerte SB, Cantoni F. Life expectancy in Alzheimer’s disease (AD). Arch Gerontol Geriatr. 2009;49 (Suppl. 1):237–43. doi: https://doi.org/10.1016/j.archger.2009.09.035
2. Golbe LI, Leyton CE. Life expectancy in Parkinson disease. Neurology. 2018;91:991–2. doi: https://doi.org/10.1212/WNL.0000000000006560
3. Caligiore D, Giocondo F, Silvetti M. The neurodegenerative elderly syndrome (NES) hypothesis: Alzheimer and Parkinson are two faces of the same disease. IBRO Neurosci Rep. 2022;13:330–43. doi: https://doi.org/10.1016/j.ibneur.2022.09.007
4. Chen JJ.Parkinson’s disease: health-related quality of life, economic cost, and implications of early treatment. Am J Manag Care. 2010;16 (Suppl):S87–93.
5. del Campo M, Peeters CFW, Johnson ECB, Vermunt L, Hok-A-Hin YS, van Nee M, et al. CSF proteome profiling across the Alzheimer’s disease spectrum reflects the multifactorial nature of the disease and identifies specific biomarker panels. Nat Aging. 2022;2:1040–53. doi: https://doi.org/10.1038/s43587-022-00300-1
6. G?tarek P, Ka?u?na-Czapli?ska J.Integrated metabolomics and proteomics analysis of plasma lipid metabolism in Parkinson’s disease. Expert Rev Proteom. 2024;21:13–25. doi: https://doi.org/10.1080/14789450.2024.2315193
7. Van Bulck M, Sierra-Magro A, Alarcon-Gil J, Perez-Castillo A, Morales-Garcia JA. Novel approaches for the treatment of Alzheimer’s and Parkinson’s disease. Int J Mol Sci. 2019;20:719. doi: https://doi.org/10.3390/ijms20030719
8. Lane RM, Potkin SG, Enz A. Targeting acetylcholinesterase and butyrylcholinesterase in dementia. Int J Neuropsychopharmacol. 2006;9:101–24. doi: https://doi.org/10.1017/S1461145705005833
9. Schliebs R, Arendt T. The cholinergic system in aging and neuronal degeneration. Behav Brain Res. 2011;221:555–63. doi: https://doi.org/10.1016/j.bbr.2010.11.058
10. Ballard CG, Greig NH, Guillozet-Bongaarts AL, Enz A, Darvesh S. Cholinesterases: roles in the brain during health and disease. Curr Alzheimer Res. 2005;2:307–18. doi: https://doi.org/10.2174/1567205054367838
11. Maiuolo J, Costanzo P, Masullo M, D’Errico A, Nasso R, Bonacci S, et al. Hydroxytyrosol-donepezil hybrids play a protective role in an in vitro induced Alzheimer’s disease model and in neuronal differentiated human SH-SY5Y neuroblastoma cells. Int J Mol Sci. 2023;24:13461. doi: https://doi.org/10.3390/ijms241713461
12. d’Angremont E, Begemann MJH, van Laar T, Sommer IEC. Cholinesterase inhibitors for treatment of psychotic symptoms in Alzheimer disease and Parkinson disease: a meta-analysis. JAMA Neurol. 2023;80:813–23. doi: https://doi.org/10.1001/jamaneurol.2023.1835
13. Azargoonjahromi A. The duality of amyloid-β: its role in normal and Alzheimer’s disease states. Mol Brain. 2024;17:44. doi: https://doi.org/10.1186/s13041-024-01118-1
14. Pagano K, Tomaselli S, Molinari H, Ragona L. Natural compounds as inhibitors of Aβ peptide aggregation: chemical requirements and molecular mechanisms. Front Neurosci. 2020;14:619667. doi: https://doi.org/10.3389/fnins.2020.619667
15. Hansen RA, Gartlehner G, Webb AP, Morgan LC, Moore CG, Jonas DE. Efficacy and safety of donepezil, galantamine, and rivastigmine for the treatment of Alzheimer’s disease: a systematic review and meta-analysis. Clin Interv Aging. 2008;3:211–25.
16. Kumar A, Tiwari A, Sharma A. Changing paradigm from one target one ligand towards multi-target directed ligand design for key drug targets of Alzheimer disease: an important role of in silico methods in multi-target directed ligands design. Curr Neuropharmacol. 2018;16:726–39. doi: https://doi.org/10.2174/1570159X16666180315141643
17. Campos-Esparza MR, Torres-Ramos MA. Neuroprotection by natural polyphenols: molecular mechanisms. Cent Nerv Syst Agents Med Chem. 2010;10:269–77. doi: https://doi.org/10.2174/187152410793429728
18. Nasso R, Pagliara V, D’Angelo S, Rullo R, Masullo M, Arcone R. Annurca apple polyphenol extract affects acetyl-cholinesterase and mono-amine oxidase in vitro enzyme activity. Pharmaceuticals (Basel). 2021;14:62. doi: https://doi.org/10.3390/ph14010062
19. Silva RFM, Poga?nik L. Polyphenols from food and natural products: neuroprotection and safety. Antioxidants (Basel). 2020;9:61. doi: https://doi.org/10.3390/antiox9010061
20. Arias-Sánchez RA, Torner L, Fenton Navarro B. Polyphenols and neurodegenerative diseases: potential effects and mechanisms of neuroprotection. Molecules. 2023;28:5415. doi: https://doi.org/10.3390/molecules28145415
21. Ullah A, Munir S, Badshah SL, Khan N, Ghani L, Poulson BG, et al. Important flavonoids and their role as a therapeutic agent. Molecules. 2020;25:5243. doi: https://doi.org/10.3390/molecules25225243
22. Arcone R, D’Errico A, Nasso R, Rullo R, Poli A, Di Donato P, et al. Inhibition of enzymes involved in neurodegenerative disorders and Aβ1-40 aggregation by Citrus limon peel polyphenol extract. Molecules. 2023;28:6332. doi: https://doi.org/10.3390/molecules28176332
23. Orhan I, Aslan S, Kartal M, ?ener B, Hüsnü Can Ba?er K. Inhibitory effect of Turkish Rosmarinus officinalis L. on acetylcholinesterase and butyrylcholinesterase enzymes. Food Chem. 2008;108:663–668. doi: https://doi.org/10.1016/j.foodchem.2007.11.023
24. de Torre MP, Cavero RY, Calvo MI. Anticholinesterase activity of selected medicinal plants from Navarra region of Spain and a detailed phytochemical investigation of Origanum vulgare L. ssp. vulgare. Molecules. 2022;27:7100. doi: https://doi.org/10.3390/molecules27207100
25. Tava A, Biazzi E, Ronga D, Pecetti L, Avato P. Biologically active compounds from forage plants. Phytochem Rev. 2022;21:471–501. doi: https://doi.org/10.1007/s11101-021-09779-9
26. Usman M, Khan WR, Yousaf N, Akram S, Murtaza G, Kudus KA, et al. Exploring the hytochemicals and anti-cancer potential of the members of Fabaceae family: a comprehensive review. Molecules. 2022;27:3863. doi: https://doi.org/10.3390/molecules27123863
27. Burlando B, Pastorino G, Salis A, Damonte G, Clericuzio M, Cornara L. The bioactivity of Hedysarum coronarium extracts on skin enzymes and cells correlates with phenolic content. Pharm Biol. 2017;55:1984–91. doi: https://doi.org/10.1080/13880209.2017.1346 691
28. Bora KS, Sharma A. Evaluation of antioxidant and cerebroprotective effect of Medicago sativa Linn. against ischemia and reperfusion insult. Evid Based Complement Alternat Med. 2011;2011:792167. doi: https://doi.org/10.1093/ecam/neq019
29. Raeeszadeh M, Mortazavi P, Atashin-Sadafi R. The ntioxidant, anti-inflammatory, pathological, and behavioural effects of Medicago sativa L. (Alfalfa) extract on brain injury caused by nicotine in male rats. Evid Based Complement Alternat Med. 2021;2021:6694629. doi: https://doi.org/10.1155/2021/6694629
30. Rolnik A, Olas B. The plants of the Asteraceae family as agents in the protection of human health. Int J Mol Sci. 2021;22:3009. doi: https://doi.org/10.3390/ijms22063009
31. Perovi? J, Tumbas Šaponjac V, Koji? J, Krulj J, Moreno DA, García-Viguera C et al. Chicory (Cichorium intybus L.) as a food ingredient -Nutritional composition, bioactivity, safety, and health claims: a review. Food Chem. 2021;336:127676. doi: https://doi.org/10.1016/j.foodchem.2020.127676
32. Rullo R, Nasso R, D’Errico A, Biazzi E, Tava A, Landi N, et al. Effect of polyphenolic extracts from leaves of Mediterranean forage crops of enzymrs involved in the oxidative stress, and useful for alternative cancer treatments. J Appl Pharm Sci. 2025;15:110–20. doi: https://doi.org/10.7324/JAPS.2025.235772
33. Scacco S, Acquaviva S, França Vieira e Silva F, Zhang JH, Lo Muzio L, Corso G, et al. Bioactivity and neuroprotective effects of extra virgin olive oil in a mouse model of cerebral ischemia: an in vitro and in vivo study. Int J Mol Sci. 2025;26:1771–802. https://doi.org/10.3390/ijms26041771
34. Vitale RM, Morace AM, D’Errico A, Ricciardi F, Fusco A, Boccella S, et al. Identification of cannabidiolic and cannabigerolic acids as MTDL AChE, BuChE, and BACE-1 inhibitors against Alzheimer’s disease by in silico, in vitro, and in vivo studies. Phytother Res. 2025;39:233–45. doi: https://doi.org/10.1002/ptr.8369
35. Rullo R, Cerchia C, Nasso R, Romanelli V, De Vendittis E, Masullo M, et al. Novel reversible inhibitors of xanthine oxidase targeting the active site of the enzyme. Antioxidants (Basel). 2023;12:825. doi: https://doi.org/10.3390/antiox12040825
36. Jalouli M, Rahman MA, Biswas P, Rahman H, Harrath AH, Lee I-S, et al. Targeting natural antioxidant polyphenols to protect neuroinflammation and neurodegenerative diseases: a comprehensive review. Front Pharmacol. 2025;16:1492517. doi: https://doi.org/10.3389/fphar.2025.1492517
37. Carregosa D, Mota S, Ferreira S, Alves-Dias B, Loncarevic-Vasiljkovic N, Crespo CL, et al. Overview of beneficial effects of (poly)phenol metabolites in the context of neurodegenerative diseases on model organisms. Nutrients. 2021;13:2940. doi: https://doi.org/10.3390/nu13092940
38. Spencer JPE. Food for thought: the role of dietary flavonoids in enhancing human memory, learning and neuro-cognitive performance. Proc Nutr Soc. 2008;67:238–52. doi: https://doi.org/10.1017/S0029665108007088
39. Psaltopoulou T, Sergentanis TN, Panagiotakos DB, Sergentanis IN, Kosti R, Scarmeas N. Mediterranean diet, stroke, cognitive impairment and depression: a meta-analysis. Ann Neurol. 2013;74:580–91. doi: https://doi.org/10.1002/ana.23944
40. Mayr HL, Thomas CJ, Tierney AC, Kucianski T, George ES, Ruiz-Canela M, et al. Randomization to 6-month Mediterranean diet compared with a low-fat diet leads to improvement in the dietary inflammatory index scores in patients with coronary heart disease: The AUSMED heart trial. Nutr Res. 2018;55:94–107. doi: https://doi.org/10.1016/j.nutres.2018.04.006
41. Li F, Zhan C, Dong X, Wei G. Molecular mechanisms of resveratrol and EGCG in the inhibition of Aβ42 aggregation and disruption of Aβ42 protofibril: similarities and differences. Phys Chem Chem Phys. 2021;23:18843–54. doi: https://doi.org/10.1039/d1cp01913a
42. Sun Y, Wang X, Zhang X, Li Y, Wang D, Sun F, et al. Di-caffeoylquinic acid: a potential inhibitor for amyloid-beta aggregation. J Nat Med. 2024;78:1029–43. doi: https://doi.org/10.1007/s11418-024-01825-y
43. El-Saadony MT, Yang T, Saad AM, Alkafaas SS, Elkafas SS, Eldeeb GS, et al. Polyphenols: Chemistry, bioavailability, bioactivity, nutritional aspects and human health benefits: a review. Int Biol Macromol. 2024;277:134223. doi: https://doi.org/10.1016/j.ijbiomac.2024.134223
44. Sun S, Liu Z, Lin M, Gao N, Wang X. Polyphenols in health and food processing: antibacterial, anti-inflammatory, and antioxidant insights. Front Nutr. 2024;11:1456730. doi: https://doi.org/10.3389/fnut.2024.1456730
45. Dubey SK, Ram MS, Krishna KV, Saha RN, Singhvi G, Agrawal M, et al. Recent expansions on cellular models to uncover the scientific barriers towards drug development for Alzheimer’s disease. Cell Mol Neurobiol. 2019;39:181–209. doi: https://doi.org/10.1007/s10571-019-00653-z
Year
Month