Isolation and Characterization of Acetylcholinesterase Inhibitors from Piper longum and Binding Mode Predictions^{[ # ]}

Abstract

Restoration of cholinergic function is considered a rational approach to enhance cognitive performance. Acetylcholinesterase inhibitors are still the best therapeutic option for Alzheimerʼs disease. The fruits of Piper longum have been used in traditional medicines for the treatment of memory loss. It was demonstrated that the dichloromethane extract of these fruits is able to inhibit acetylcholinesterase. Thus, the aim of this study was to identify the contained acetylcholinesterase inhibitors. The active zones were presented via TLC-bioautography, and five compounds were isolated in the process of a bioassay-guided phytochemical investigation. Their structures were characterized as piperine, methyl piperate, guineenisine, pipercide, and pellitorine using spectroscopy and spectrometry methods (UV, IR, MS, ¹H-, and ¹³C-NMR). In vitro acetylcholinesterase inhibitory activities of the isolates and their IC₅₀ values were determined via a colorimetric assay. Three of them exhibited enzyme inhibitory activities, with piperine being the most potent compound (IC₅₀ of 0.3 mM). In order to investigate the binding mode of the tested compounds, docking studies were performed using the X-ray crystal structure of acetylcholinesterase from Tetronarce californica with the Protein Data Bank code 1EVE. The content of the active compounds in the extract was determined by a developed HPLC method. Piperine was present in the maximum quantity in the fruits (0.57%), whereas methyl piperate contained the minimum content (0.10%).

Key words

Alzheimerʼs disease - AChE inhibitor - Piper longum - Piperaceae - piperine - pipercide - guineensine

^# This article is dedicated to Prof. Wolfgang Kubelka for a lifetime of effort in the field of Pharmacognosy.

Publication History

Received: 22 January 2020

Accepted after revision: 13 June 2020

Article published online:
15 July 2020

• References

• 1 Hugo J, Ganguli M. Dementia and cognitive impairment: epidemiology, diagnosis, and treatment. Clin Geriatr Med 2014; 30: 421-442
• 2 Wimo A, Jonsson L, Bond J, Prince M, Winblad B. The worldwide economic impact of dementia 2010. Alzheimerʼs & dementia. Alzheimers Dement 2013; 9: 1-11
• 3 Bhounsule AS, Bhatt LK, Prabhavalkar KS, Oza M. Cyclin dependent kinase 5: A novel avenue for Alzheimerʼs disease. Brain Res Bull 2017; 132: 28-38
• 4 Eldufani J, Blaise G. The role of acetylcholinesterase inhibitors such as neostigmine and rivastigmine on chronic pain and cognitive function in aging: A review of recent clinical applications. Alzheimers Dement (NY) 2019; 5: 175-183
• 5 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: 770-803
• 6 Hostettmann K, Borloz A, Urbain A, Marston A. Natural product inhibitors of acetylcholinesterase. Curr Org Chem 2006; 10: 825-847
• 7 Schuster D, Kern L, Hristozov DP, Terfloth L, Bienfait B, Laggner C, Kirchmair J, Grienke U, Wolber G, Langer T, Stuppner H, Gasteiger J, Rollinger JM. Applications of integrated data mining methods to exploring natural product space for acetylcholinesterase inhibitors. Comb Chem High Throughput Screen 2010; 13: 54-66
• 8 Kritikar KR, Basu BD. Indian medicinal Plants. Mumbai: Orients Longman; 1980: 21-28
• 9 Dwivedi C, Chandrakar K, Singh V, Tiwari SP, Satapathy T, Kesharwani S, Kumar B, Roy A. Indian herbal medicines used for treatment of dementia: an overview. Int J Pharmacogn 2014; 1: 553-571
• 10 Ahmadian-Attari MM, Ahmadiani A, Kamalinejad M, Dargahi L, Shirzad M, Mosaddegh M. Treatment of Alzheimerʼs disease in Iranian traditional medicine. Iran Red Crescent Med J 2014; 17: e18052
• 11 Yadav V, Krishnan A, Vohora D. A systematic review on Piper longum L.: Bridging traditional knowledge and pharmacological evidence for future translational research. J Ethnopharmacol 2020; 247: 112255
• 12 Adhami HR, Farsam H, Krenn L. Screening of medicinal plants from Iranian traditional medicine for acetylcholinesterase inhibition. Phytother Res 2011; 25: 1148-1152
• 13 Gómez-Calvario V, Rios MY. ¹H and ¹³C NMR data, occurrence, biosynthesis, and biological activity of Piper amides. Magn Reson Chem 2019; 57: 994-1070
• 14 Lee SA, Hwang JS, Han XH, Lee C, Lee MH, Choe SG, Hong SS, Lee D, Lee MK, Hwang BY. Methylpiperate derivatives from Piper longum and their inhibition of monoamine oxidase. Arch Pharm Res 2008; 31: 679-683
• 15 Ee GC, Lim CM, Rahmani M, Shaari K, Bong CF. Pellitorine, a potential anti-cancer lead compound against HL6 and MCT-7 cell lines and microbial transformation of piperine from Piper nigrum . Molecules 2012; 15: 2398
• 16 GOLD 5.2, The Cambridge Crystallographic Data Centre. Available at (Accessed November 2, 2018): https://www.ccdc.cam.ac.uk
• 17 Jones G, Willett P, Glen RC, Leach AR, Taylor R. Development and validation of a genetic algorithm for flexible docking. J Mol Biol 1997; 267: 727-748
• 18 Karan RS, Bhargava VK, Garg SK. Effect of piperine on the pharmacokinetic profile of isoniazid in rabbits. Indian J Pharmacol 1998; 30: 254-256
• 19 Kapoor IPS, Singh B, Singh G, De Heluani CS, De Lampasona MP, Catalan CAN. Chemistry and in vitro antioxidant activity of volatile oil and oleoresins of Black pepper (Piper nigrum). J Agric Food Chem 2009; 57: 5358-5364
• 20 Rahman T, Rahmatullah M. Proposed structural basis of interaction of piperine and related compounds with monoamine oxidases. Bioorg Med Chem Lett 2010; 20: 537-540
• 21 Okello EJ, Coleman A, Seal CJ. In-Vitro anti-cholinesterase activities by piperine, an alkaloid from the spice family piperaceae. Int J Pharm Sci Res 2015; 6: 3726-3732
• 22 Yanbei T, Yujiao Z, Hongjian D, Wei L, Yaya W, Qin L, Chao Z, Yanfang L. Anticholinesterases and antioxidant alkamides from Piper nigrum fruits. Nat Prod Res 2016; 30: 1945-1949
• 23 Kong LD, Christopher HK, Cheng RX. Inhibition of MAO A and B by some plant-derived alkaloids, phenols and anthraquinones. J Ethnopharmacol 2004; 91: 351-355
• 24 Qu H, Yu X, Lian X, Xu H. Discovery of some piperine-based phenylsulfonylhydrazone derivatives as potent botanically narcotic agents. Sci Rep 2015; 5: 13077
• 25 Nicolussia S, Paredesb JM, Gacheta MS, Raua M, Sotob ME, Blunderc M, Gertsch J. Guineensine is a novel inhibitor of endocannabinoid uptake showing cannabimimetic behavioral effects in BALB/c mice. Pharmacol Res 2014; 80: 52-65
• 26 Hutter-Paier B, Huttunen H, Puglielli L, Eckman C, Kim D, Hofmeister AD, Moir R, Windisch M. The ACAT inhibitor CP-113, 818 markedly reduces amyloid pathology in a mouse model of Alzheimerʼs disease. Neuron 2004; 44: 227-238
• 27 Huttunen HJ, Kovacs DM. ACAT as a drug target for Alzheimerʼs disease. Neurodegener Dis 2008; 5: 212-214
• 28 Rho MV, Lee SW, Park HR, Choi J, Kang Y, Kim K. ACAT inhibition of alkamides identified in the fruits of Piper nigrum . Phytochemistry 2007; 68: 899-903
• 29 Adhami HR, Lutz J, Kahlig H, Zehl M, Krenn L. Compounds from gum ammoniacum with acetylcholinesterase inhibitory activity. Sci Pharm 2013; 81: 793-805
• 30 Kryger G, Sillman I, Sussman JL. Structure of acetylcholinesterase complexed with E2020 (Aricept): implications for the design of new anti-Alzheimer drugs. Structure 1999; 7: 297-307
• 31 Bajda M, Więckowska A, Hebda M, Guzior N, Sotriffer CA, Malawska B. Structure-based search for new inhibitors of cholinesterases. Int J Mol Sci 2013; 14: 5608-5632
• 32 Hawkins PCD, Skillman AG, Warren GL, Ellingson BA, Stahl MT. Omega 2.5.1.4. OpenEye Scientific Software. Available at (Accessed November 2, 2018): http://www.eyesopen.com
• 33 Hawkins PCD, Skillman AG, Warren GL, Ellingson BA, Stahl MT. Conformer generation with OMEGA: algorithm and validation using high quality structures from the Protein Databank and the Cambridge Structural Database. J Chem Inf Model 2010; 50: 572-584
• 34 Discovery Studio 4.0, Dassault Systèmes. Available at (Accessed November 2, 2018): http://www.3ds.com/de
• 35 Wolber G, Adaktylos P, Bendix F, Biely M, Dornhofer A, Gökhan I, Kosara R, Seidel T. LigandScout 3.12. Inte: Ligand GmbH; 2018. Available at (Accessed November 2) http://www.inteligand.com
• 36 Adhami HR, Fitz V, Lubich A, Kaehlig H, Zehl M, Krenn L. Acetylcholinesterase inhibitors from galbanum, the oleo gum-resin of Ferula gummosa Boiss. Phytochem Lett 2014; 10: 32-87