• VBSC Thunuguntla
  • Kala Kumar B
  • BONDILI JS K L University


Objective: Current drugs to Alzheimer's disease (AD) are targeted to delay the breakdown of acetylcholine, thereby increasing the concentration
of acetylcholine released into synaptic cleft and enhancing cholinergic neurotransmission. This paper deals with screening and identification of
acetylcholinesterase (AChE) inhibitors in solvent extracts of Hyptis suaveolens (HS).
Methods: In search of natural inhibitors of AChE, this study is focused on extract of HS, a member of Lamiaceae. 1:4 ratio of methanolic extract
is prepared with shade dried areal parts of HS plant. The extract was assayed by Ellman's method for inhibition activity and then purified using
ammonium sulfate precipitation and chromatography techniques. Gas chromatography-mass spectrometry (GC-MS) identified compounds were
analyzed by docking studies.
Results: Methanolic extract showed maximum percentage inhibition of 75.00±4.30 (2.1 mg/ml) with an IC
value of 1.020±0.026 mg/ml.
However, saturated ammonium sulfate precipitation of methanolic extract and further fractionation by gel permeation chromatography showed
86.00±1.30% AChE inhibition (AChEI) activity. Reverse phase high-performance liquid chromatography (RP-HPLC) fraction (retention time [RT]
5.170) showed significant inhibition when compared to the other peak (RT 6.643). RP-HPLC fraction (RT 5.170) with significant inhibition was
identified as Eugenol by GC-MS analysis. In silico analysis of all the GC-MS identified molecules revealed Eugenol as possessing preeminent absorption,
distribution, metabolism, elimination properties and a glide (docking) score of −9.14 kcal/mole with AChE enzyme of pacific electric ray (Torpedo
californica - TcAChE) (PDB ID: 1EVE).
Conclusion: Screening, purification and identification, and identification of diverse phytochemicals of the HS plant, as potent source of AChEI.
Keywords: Acetylcholinesterase, Acetylcholnesterase inhibitor, Hyptis suaveolens, Alzheimer's disease, Dementia.


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. Johnson N, Davis T, Bosanquet N. The epidemic of Alzheimer’s

disease. How can we manage the costs? Pharmacoeconomics


Wimo A, Winblad B, Jönsson L. The worldwide societal costs of

dementia: Estimates for 2009. Alzheimers Dement 2010;6(2):98-103.

The WHO Report Dementia. Available from:

[Last accessed on 2015 Mar].

Krall WJ, Sramek JJ, Cutler NR. Cholinesterase inhibitors: A therapeutic

strategy for Alzheimer disease. Ann Pharmacother 1999;33(4):441-50.

Giacobini E. Cholinergic foundations of Alzheimer’s disease therapy.

J Physiol Paris 1998;92(3-4):283-7.

Easwaramoorthy B, Pichika R, Collins D, Potkin SG, Leslie FM,

Mukherjee J. Effect of acetylcholinesterase inhibitors on the binding

of nicotinic alpha-4 beta-2 receptor PET radiotracer, (18) F-nifene:

A measure of acetylcholine competition. Synapse 2007;61(1):29-36.

Qizilbash N, Whitehead A, Higgins J, Wilcock G, Schneider L,

Farlow M. Cholinesterase inhibition for Alzheimer disease: A

meta-analysis of the tacrine trials. Dementia trialists’ collaboration.

JAMA 1998;280(20):1777-82.

Arrieta JL, Artalejo FR. Methodology, results and quality of clinical

trials of tacrine in the treatment of Alzheimer’s disease: A systematic

review of the literature. Age Ageing 1998;27(2):161-79.

Jones RW, Soininen H, Hager K, Aarsland D, Passmore P, Murthy A,

et al. A multinational, randomised, 12-week study comparing the

effects of donepezil and galantamine in patients with mild to moderate

Alzheimer’s disease. Int J Geriatr Psychiatry 2004;19(1):58-67.

Polinsky RJ. Clinical pharmacology of rivastigmine: A new-generation

acetylcholinesterase inhibitor for the treatment of Alzheimer’s disease.

Clin Ther 1998;20(4):634-47.

Castro DM, Dillon C, Machnicki G, Allegri RF. The economic cost of

Alzheimer’s disease. Dement Neuropsychol 2010;4 Suppl 4:262-7.

Sofowora A. Research on medicinal plants and traditional medicine in

Africa. J Altern Complement Med 1996;2(3):365-72.

Robbers J, Speedie M, Tyler V. Pharmacognosy and

Pharmacobiotechnology. Baltimore, USA: Williams and Wilkins; 1996.

Kingston DG, Rao MM, Zucker WV. Plant anticancer agents. IX.

Constituents of Hyptis tomentosa. J Nat Prod 1979;42(5):496-9.

Edeoga H, Omosun G, Uche L. Chemical composition of Hyptis

suaveolens and Ocimum gratissimum hybrids from Nigeria. Afr J

Biotechnol 2006;5 Suppl 10:892-5.

Das PK, Sahoo S, Sethi R, Nayak PS, Nayak S, Joshi A. Phytochemical

and pharmacological investigation of the protective effect of plant

Hyptis suaveolens against duodenal ulceration. J Glob Pharm Technol

;1 Suppl 1:99-103.

Shirwaikar A, Shenoy R, Udupa AL, Udupa SL, Shetty S. Wound

healing property of ethanolic extract of leaves of Hyptis suaveolens

with supportive role of antioxidant enzymes. Indian J Exp Biol


Nantitanon W, Chowwanapoonpohn S, Okonogi S. Antioxidant and

antimicrobial activities of Hyptis suaveolens essential oil. Sci Pharm

;75 Suppl 1:35.

Ren Y, Houghton PJ, Hider RC, Howes MJ. Novel diterpenoid

acetylcholinesterase inhibitors from Salvia miltiorhiza. Plant Med

;70 Suppl 3:201-4.

Ellman GL, Courtney KD, Andres V Jr, Feather-Stone RM. A new

and rapid colorimetric determination of acetylcholinesterase activity.

Biochem Pharmacol 1961;7:88-95.

Parekh J, Chanda SV. In vitro antimicrobial activity and

phytochemical analysis of some Indian medicinal plants. Turk J Biol

;31 Suppl 1:53-8.

Raghavendra M, Satish S, Raveesha K. Phytochemical analysis and

antibacterial activity of Oxalis corniculata; a known medicinal plant.

Science 2006;1 Suppl 1:72-8.

Harborne JB. Phytochemical Methods: A Guide to Modern Techniques

of Plant Analysis. New Delhi: Springer; 1998.

Mukherjee PK, Kumar V, Mal M, Houghton PJ. Acetylcholinesterase

inhibitors from plants. Phytomedicine 2007;14(4):289-300.

Orhan I, Terzioglu S, Sener B. Alpha-onocerin: An acetylcholinesterase

inhibitor from Lycopodium clavatum. Planta Med 2003;69(3):265-7.

Sener B, Orhan I. Discovery of drug candidates from some

Turkish plants and conservation of biodiversity. Pure Appl Chem


Barceloux DG. Medical Toxicology of Natural Substances: Foods,

Fungi, Medicinal Herbs, Plants and Venomous Animals. Hoboken, NJ,

USA: Wiley; 2008.

Asian J Pharm Clin Res, Vol 9, Issue 3, 2016, 148-153

Thunuguntla et al.

Kamatou GP, Vermaak I, Viljoen AM. Eugenol – From the remote

Maluku Islands to the international market place: A review of a

remarkable and versatile molecule. Molecules 2012;17(6):6953-81.

Wright DA, Payne JP. A clinical study of intravenous anaesthesia with

a eugenol derivative, G.29.505. Br J Anaesth 1962;34:379-85.

Zheng GQ, Kenney PM, Lam LK. Sesquiterpenes from clove (Eugenia

caryophyllata) as potential anticarcinogenic agents. J Nat Prod


Orhan I, Sener B, Choudhary MI, Khalid A. Acetylcholinesterase and

butyrylcholinesterase inhibitory activity of some Turkish medicinal

plants. J Ethnopharmacol 2004;91(1):57-60.

Adsersen A, Gauguin B, Gudiksen L, Jäger AK. Screening of

plants used in Danish folk medicine to treat memory dysfunction

for acetylcholinesterase inhibitory activity. J Ethnopharmacol


Khan RA, Bukhari IA, Nawaz SA, Choudhary MI. Acetylcholinesterase

and butyrylcholinesterase inhibitory potential of some Pakistani

medicinal plants. J Basic Appl Sci 2006;2:7-10.



How to Cite

Thunuguntla, V. ., C. S. . B, K. K. B, and B. . JS. “SCREENING AND IN SILICO ANALYSIS OF HYPTIS SUAVEOLENS METABOLITES FOR ACETYLCHOLINESTERASE INHIBITION”. Asian Journal of Pharmaceutical and Clinical Research, vol. 9, no. 3, May 2016, pp. 148-53,



Original Article(s)