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Licensed Unlicensed Requires Authentication Published by De Gruyter September 22, 2017

Modulatory effect of caffeic acid on cholinesterases inhibitory properties of donepezil

  • Odunayo Michael Agunloye EMAIL logo and Ganiyu Oboh



Donepezil hydrochloride commonly used in the management of Alzheimer’s disease (AD), exhibiting its inhibitory effects on acetylcholinesterase and butyrylcholinesterase activity thereby enhance cognitive function. Caffeic acid member of hydroxycinnamic acid is widely present in human diet. This study aims to investigate influence of caffeic acid on acetylcholinesterase and butyrylcholinesterase inhibitory properties of donepezil (in vitro).


5 mg of donepezil was dissolved in 50 mL distilled water while 10 mg of caffeic acid was dissolved in 100 mL distilled water. Therefore, mixtures of samples were prepared as follows: A2=donepezil 0.075 mg/mL+caffeic acid 0.025 mg/mL; A3=donepezil 0.050 mg/mL+caffeic acid 0.050 mg/mL; A4=donepezil 0.025 mg/mL+caffeic acid 0.075 mg/mL. All samples were kept in the refrigerator at 4 °C for subsequent analysis.


The result showed that all the combinations show an inhibitory effect on acetylcholinesterase and butyrylcholinesterase activity in vitro, with the combination A4=donepezil 0.025 mg/mL+caffeic acid 0.075 mg/mL had significant (p<0.05) highest inhibitory effect on acetylcholinesterase and butyrylcholinesterase activity in vitro. More so, all the samples were able to prevent pro-oxidants (FeSO4 and sodium nitroprusside [SNP] ) induced lipid peroxidation in rat brain homogenate, with the combination A4=donepezil 0.025 mg/mL+caffeic acid 0.075 mg/mL and A3=donepezil 0.050 mg/mL+caffeic acid 0.050 mg/mL had highest inhibitory effect against FeSO4 and SNP induced lipid peroxidation in rat brain homogenate in vitro respectively. Moreover, all the samples exhibit antioxidant properties as typified by their ability to chelate iron (II) ion (Fe2+), hydroxyl radical (OH٭) radical scavenging ability and ferric reducing property (FRAP).


Therefore, the combination of caffeic acid with donepezil enhances the antioxidant properties of donepezil. The combination of caffeic acid with donepezil could be a therapeutic aid in the management of AD, possibly with fewer side effects of donepezil. Nevertheless, the combination donepezil 0.025 mg/mL+caffeic acid 0.075 mg/mL acid look promising.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


[1] Jivad N, Rabiel Z. A review study on medicinal plants used in the treatment of learning and memory impairments. Asian Pac J Trop Biomed. 2014;4:780–9.10.12980/APJTB.4.2014APJTB-2014-0412Search in Google Scholar

[2] Ballard CG, Greig NH, Guillozet-Bongaarts AL, Enz A, Darvesh S. Cholinesterases: roles in the brain during health and disease. Curr Alzheimer Res. 2005;2:307–18.10.2174/1567205054367838Search in Google Scholar PubMed

[3] Anglister L, Etlin A, Finkel E, Durrant AR, LevTov A. Cholinesterases in development and disease. Chemico Biological Interactions. 2008;175:92–100.10.1016/j.cbi.2008.04.046Search in Google Scholar PubMed

[4] Giacobini E. Long-term stabilizing effect of cholinesterase inhibitors in the therapy of Alzheimer’ disease. J Neural Transm. 2002;62:181–7.10.1007/978-3-7091-6139-5_17Search in Google Scholar

[5] Wszelaki N, Kuciun A. Screening of traditional European herbal medicines for acetylcholinesterase and butyrylcholinesterase inhibitory activity. Acta Pharm. 2010;60:119–28.10.2478/v10007-010-0006-ySearch in Google Scholar PubMed

[6] Chaiyana W, Okonogi S. Inhibition of cholinesterase by essential oil from food plant. Phytomed. 2012;19:836–9.10.1016/j.phymed.2012.03.010Search in Google Scholar

[7] Schneider LS. Donepezil in Alzheimer’s disease. Lancet. 2004;363:2100–1.10.1016/S0140-6736(04)16533-1Search in Google Scholar PubMed

[8] Markesbery WR. Oxidative stress hypothesis in Alzheimer’s disease. Free Radic Biol Med. 1997;23:134–47.10.1016/S0891-5849(96)00629-6Search in Google Scholar PubMed

[9] Pappolla MA, Chyan YJ, Omar RA, Hsiao K, Perry G, Smith MA, Evidence of oxidative stress and in vivo neurotoxicity of β-amyloid in a transgenic mouse model of Alzheimer’s disease. A chronic oxidative paradigm for testing antioxidant therapies in vivo. Am J Pathol. 1998;152:871–7.Search in Google Scholar

[10] Kanski J, Aksenova M, Stoyanova A, Butterfield DA. Ferulic acid antioxidant protection against hydroxyl and peroxyl radical oxidation in synaptosomal and neuronal cell culture systems in vitro: structure-activity studies. J Nutr Biochem. 2002;13:273–81.10.1016/S0955-2863(01)00215-7Search in Google Scholar PubMed

[11] Doody RS. Clinical benefits of a new piperidine-class Ache inhibitor. Eur Neuro-Psychopharmacol. 1999;9:69–77.10.1016/S0924-977X(98)00047-9Search in Google Scholar

[12] Doody RS. Clinical profile of donepezil in the treatment of Alzheimer’s Disease. Gerontology. 1999;45:23–32.10.1159/000052761Search in Google Scholar PubMed

[13] Doody RS. Refining treatment guidelines in Alzheimer’s disease. Geriatrics. 2005;Suppl:14–20.Search in Google Scholar PubMed

[14] Oboh G, Rocha JBT. Distribution and antioxidant activity of polyphenols in ripe and unripe tree pepper (Capsicum pubescens). J Food Biochem. 2007;31:456–73.10.1111/j.1745-4514.2007.00123.xSearch in Google Scholar

[15] Yen HF, Hsieh CT, Hsieh TJ, Chang FR, Wang CK. In vitro antidiabetic effect and chemical component analysis of 29 essential oil products. J Food Drug Anal. 2015;3:124–9.10.1016/j.jfda.2014.02.004Search in Google Scholar

[16] Oboh G, Bello FO, Ademosun AO. Hypocholesterolemic properties of grapefruit (Citrus paradisii) and shaddock (Citrus maxima) juices and inhibition of angiotensin-1-converting enzyme activity. J Food Drug Anal. 2014;22:477–84.10.1016/j.jfda.2014.06.005Search in Google Scholar PubMed

[17] Dai Q, Borenstein AR, Wu Y, Jackson JC, Larson EB. Fruit and vegetable juices and Alzheimer’s disease: the Kame project. Am J Med. 2006;119:751–9.10.1016/j.amjmed.2006.03.045Search in Google Scholar PubMed

[18] Clifford MN. Chlorogenic acids and other cinnamatess nature, occurrence and dietary burden. J Sci Food Agric. 1999;79:362–72.10.1002/(SICI)1097-0010(19990301)79:3<362::AID-JSFA256>3.0.CO;2-DSearch in Google Scholar

[19] Olthof MR, Hollman PCH, Katan MB. Chlorogenic acid and caffeic acid are absorbed in humans. J Nutr. 2001;131:66–71.10.1093/jn/131.1.66Search in Google Scholar PubMed

[20] Oboh G, Agunloye OM, Akinyemi AJ, Ademiluyi AO, Adefegha AS. Comparative study on the inhibitory effect of caffeic and chlorogenic acids on key enzymes linked to Alzheimer’s disease and some pro-oxidant induced oxidative stress in Rats’ Brain-In Vitro. Neurochem Res. 2013;38:413–9.10.1007/s11064-012-0935-6Search in Google Scholar PubMed

[21] Ellman GL, Courtney KD, Andres V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88–95.10.1016/0006-2952(61)90145-9Search in Google Scholar PubMed

[22] Belle NAV, Dalmolin GD, Fonini G, Rubim MA, Rocha JBT. Polyamines reduces lipid peroxidation induced by different pro-oxidant agents. Brain Res. 2004;1008:245–51.10.1016/j.brainres.2004.02.036Search in Google Scholar PubMed

[23] Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95:351–8.10.1016/0003-2697(79)90738-3Search in Google Scholar PubMed

[24] Oyaizu M. Studies on products of browning reaction: Antioxidative activity of products of browning reaction prepared from glucosamine. Jpn J Nutr. 1986;44:307–15.10.5264/eiyogakuzashi.44.307Search in Google Scholar

[25] Minotti G, Aust S. An investigation into the mechanism of citrate-Fe2+ dependent lipid peroxidation. Free Radical Bio Med. 1987;3:379–87.10.1016/0891-5849(87)90016-5Search in Google Scholar

[26] Puntel RL, Nogueira CW, Rocha JBT. Krebs cycle intermediates modulate thiobarbituric reactive species (TBARS) production in rat brain in vitro. Neurochem Res. 2005;30:225–35.10.1007/s11064-004-2445-7Search in Google Scholar PubMed

[27] Halliwell JM, Gutterigde B. The measurement and mechanism of lipid peroxidation in biological systems. Trends Biochem Sci. 1990;15:129–35.10.1016/0968-0004(90)90206-QSearch in Google Scholar PubMed

[28] Norifumi T. Donepezil in the treatment of patients with Alzheimer’s disease. Expert Rev Neurother. 2009;9:591–8.10.1586/ern.09.23Search in Google Scholar PubMed

[29] Katalinic M, Rusak G, Domacinovic Barovic J, Sinko G, Jelic D, Antolovic R, Structural aspects of flavonoids as inhibitors of human butyrylcholinesterase. Eur J Med Chem. 2010;45:186–92.10.1016/j.ejmech.2009.09.041Search in Google Scholar PubMed

[30] Orhan I, Kartal M, Tosun F, Sener B. Screening of various phenolic acids and flavonoid derivatives for their anticholinesterase potential. Z Naturforsch. 2007;62c:829–32.10.1515/znc-2007-11-1210Search in Google Scholar PubMed

[31] Alcolea-Palafoxa M, Posada-Moreno P, Ortuño-Sorianob I, Pacheco-del-Cerro JL, Martínez-Rincón C, Rodríguez-Martínez D,. Research Strategies Developed for the Treatment of Alzheimer’s Disease. Reversible and Pseudo-Irreversible Inhibitors of Acetylcholinesterase: Structure-Activity Relationships and Drug Design. Drug Design and Discovery in Alzheimer’s disease, 2004 10.1016/B978-0-12-803959-5.50008-8 426–477.10.2174/9781608058228114060010Search in Google Scholar

[32] Pulido R, Bravo L, Saura-Calixto F. Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay. J Agric Food Chem. 2000;48:3396–402.10.1021/jf9913458Search in Google Scholar PubMed

[33] Youdim MB, Bakhle YS. Monoamine oxidase: Isoforms and inhibitors in Parkinson’s disease and depressive illness. Br J Pharmacol. 2006;147:287–96.10.1038/sj.bjp.0706464Search in Google Scholar PubMed PubMed Central

[34] Oboh G, Ogunsuyi OB, Ogunbadejo MD, Adefegha AS. Influence of gallic acid on α -amylase and α -glucosidase inhibitory properties of acarbose. Jfda. 2016;24:627–34.10.1016/j.jfda.2016.03.003Search in Google Scholar PubMed

[35] Burdo JR, Connor JR. Brain iron uptake and homeostatic mechanisms. An overview. BioMetals. 2003;16:63–75.10.1023/A:1020718718550Search in Google Scholar PubMed

[36] Zecca L, Youdim MH, Riederer P, Connor J, Crichton R. Iron, brain ageing and Neurodegenerative disorders. Nat Rev Neurosci. 2004;5:863–73.10.1038/nrn1537Search in Google Scholar PubMed

Received: 2017-2-14
Accepted: 2017-6-29
Published Online: 2017-9-22

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