Bioactivity Guided Fractionation and Purification of Anti-Depressant Molecule from Ashwagandha (Withania somnifera)

Author(s): Benny Antony*, A.P.A. Aravind, Merina Benny, Nishant K. Gupta, Biji Joseph, Anu Sebastian

Journal Name: Current Bioactive Compounds

Volume 16 , Issue 5 , 2020

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Abstract:

Background: Ashwagandha (Withania somnifera) is an important herb in the Indian traditional system of medicine for neurological disorders. However, the efforts for isolation and characterisation of a molecule with anti-depressant activity and development as a potent dosage form are limited.

Objective: The objective of the present study was to characterize the Ashwagandha extract for its antidepressant fraction or constituent and to improve biological benefits at low doses.

Methods: Aqueous methanol extract of Ashwagandha was prepared and fractionated into withanolides and flavonoids rich fractions. Withanolide rich fraction was subjected to phytochemical analysis to identify the active molecule/s. The compound was purified by using a semi-preparative HPLC system; identified using various spectroscopic techniques and anti-depressant activity was evaluated in rats. Enteric coating was performed on the extract and fractions after granulation and anti-depressant activity of coated samples were evaluated in rats.

Results: Aqueous methanol extract of Ashwagandha and withanolide rich fraction showed prominent dose-dependent anti-depressant activity in forced swim test in rats. Phytochemical analysis of active fraction resulted in the isolation and characterization of a major withanolide glycoside present, namely withanoside X. Enteric coated aqueous methanol extract, withanolide rich fraction and withanoside X showed significant antidepressant activity at low doses as compared to the uncoated forms.

Conclusion: The active fraction/isolated compound is sensitive to low pH of the stomach, thus enteric coating might be beneficial to protect the actives in the stomach, facilitating the sustainable release into the intestine and in turn reduce the dosage.

Keywords: Ashwagandha, Withania somnifera, forced swim test, anti-depression, withanoside X, withanolide.

[1]
Umadevi, P.; Murugan, S.; Jennifer Suganthi, S.; Subakanmani, S. Evaluation of antidepressant like activity of Cucurbita pepo seed extracts in rats. Int. J. Curr. Pharm. Res., 2011, 3(1), 108-113.
[2]
Schechter, L.E.; Ring, R.H.; Beyer, C.E.; Hughes, Z.A.; Khawaja, X.; Malberg, J.E.; Rosenzweig-Lipson, S. Innovative approaches for the development of antidepressant drugs: current and future strategies. NeuroRx, 2005, 2(4), 590-611.
[http://dx.doi.org/10.1602/neurorx.2.4.590] [PMID: 16489368]
[3]
Belmaker, R.H.; Agam, G. Major depressive disorder. N. Engl. J. Med., 2008, 358(1), 55-68.
[http://dx.doi.org/10.1056/NEJMra073096] [PMID: 18172175]
[4]
Zhang, Z.J. Therapeutic effects of herbal extracts and constituents in animal models of psychiatric disorders. Life Sci., 2004, 75(14), 1659-1699.
[http://dx.doi.org/10.1016/j.lfs.2004.04.014] [PMID: 15268969]
[5]
Bharathi, P.; Seshayamma, V.; Jagannadharao, G.H.; Sivakumar, N. Evaluation of antidepressant activity of aqueous extract of Withania somnifera [Aswagandha] roots in albino mice. IOSR J. Pharma. Biol. Sci., 2015, 10, 27-29.
[6]
Viña, D.; Serra, S.; Lamela, M.; Delogu, G. Herbal natural products as a source of monoamine oxidase inhibitors: a review. Curr. Top. Med. Chem., 2012, 12(20), 2131-2144.
[http://dx.doi.org/10.2174/156802612805219996] [PMID: 23231392]
[7]
Kulkarni, S.K.; Bhutani, M.K.; Bishnoi, M. Antidepressant activity of curcumin: involvement of serotonin and dopamine system. Psychopharmacology (Berl.), 2008, 201(3), 435-442.
[http://dx.doi.org/10.1007/s00213-008-1300-y] [PMID: 18766332]
[8]
Irie, Y. Effects of eugenol on the central nervous system: its possible application to treatment of alzheimer’s disease, depression, and parkinson’s disease. Curr. Bioact. Compd., 2006, 2(1), 57-66.
[http://dx.doi.org/10.2174/1573407210602010057]
[9]
Kong, L.D.; Cheng, C.H.; Tan, R.X. Inhibition of MAO A and B by some plant-derived alkaloids, phenols and anthraquinones. J. Ethnopharmacol., 2004, 91(2-3), 351-355.
[http://dx.doi.org/10.1016/j.jep.2004.01.013] [PMID: 15120460]
[10]
Zhu, W.; Ma, S.; Qu, R.; Kang, D.; Liu, Y. Antidepressant effect of baicalin extracted from the root of Scutellaria baicalensis in mice and rats. Pharm. Biol., 2006, 44(7), 503-510.
[http://dx.doi.org/10.1080/13880200600878684]
[11]
Bhattacharya, S.K.; Muruganandam, A.V. Adaptogenic activity of Withania somnifera: an experimental study using a rat model of chronic stress. Pharmacol. Biochem. Behav., 2003, 75(3), 547-555.
[http://dx.doi.org/10.1016/S0091-3057(03)00110-2] [PMID: 12895672]
[12]
Bhattacharya, S.K.; Bhattacharya, A.; Sairam, K.; Ghosal, S. Anxiolytic-antidepressant activity of Withania somnifera glycowithanolides: an experimental study. Phytomedicine, 2000, 7(6), 463-469.
[http://dx.doi.org/10.1016/S0944-7113(00)80030-6] [PMID: 11194174]
[13]
Wadhwa, R.; Konar, A.; Kaul, S.C. Nootropic potential of Ashwagandha leaves: Beyond traditional root extracts. Neurochem. Int., 2016, 95, 109-118.
[http://dx.doi.org/10.1016/j.neuint.2015.09.001] [PMID: 26361721]
[14]
Jayaprakasam, B.; Zhang, Y.; Seeram, N.P.; Nair, M.G. Growth inhibition of human tumor cell lines by withanolides from Withania somnifera leaves. Life Sci., 2003, 74(1), 125-132.
[http://dx.doi.org/10.1016/j.lfs.2003.07.007] [PMID: 14575818]
[15]
Minguzzi, S.; Barata, L.E.; Shin, Y.G.; Jonas, P.F.; Chai, H.B.; Park, E.J.; Pezzuto, J.M.; Cordell, G.A. Cytotoxic withanolides from Acnistus arborescens. Phytochemistry, 2002, 59(6), 635-641.
[http://dx.doi.org/10.1016/S0031-9422(02)00022-5] [PMID: 11867095]
[16]
Habtemariam, S. Cytotoxicity and immunosuppressive activity of withanolides from Discopodium penninervium. Planta Med., 1997, 63(1), 15-17.
[http://dx.doi.org/10.1055/s-2006-957594] [PMID: 9063090]
[17]
Chandrasekaran, S.; Veronica, J.; Sundar, S.; Maurya, R. Alcoholic fractions F5 and F6 from Withania somnifera leaves show a potent antileishmanial and immunomodulatory activities to control experimental visceral Leishmaniasis. Front. Med. (Lausanne), 2017, 4, 55.
[http://dx.doi.org/10.3389/fmed.2017.00055] [PMID: 28553635]
[18]
Kuboyama, T.; Tohda, C.; Komatsu, K. Withanoside IV and its active metabolite, sominone, attenuate Abeta(25-35)-induced neurodegeneration. Eur. J. Neurosci., 2006, 23(6), 1417-1426.
[http://dx.doi.org/10.1111/j.1460-9568.2006.04664.x] [PMID: 16553605]
[19]
Shah, P.C.; Trivedi, N.A.; Bhatt, J.D.; Hemavathi, K.G. Effect of Withania somnifera on forced swimming test induced immobility in mice and its interaction with various drugs. Indian J. Physiol. Pharmacol., 2006, 50(4), 409-415.
[PMID: 17402272]
[20]
Gupta, M.; Kaur, G. Aqueous extract from the Withania somnifera leaves as a potential anti-neuroinflammatory agent: a mechanistic study. J. Neuroinflammation, 2016, 13(1), 193.
[http://dx.doi.org/10.1186/s12974-016-0650-3] [PMID: 27550017]
[21]
Jayesh, P.; Pranav, J.; Mafatlal, T. Process for extraction of ashwagandha (Withania somnifera) roots. WO2012160569A1, November 29, 2012.
[22]
Porsolt, R.D.; Bertin, A.; Jalfre, M. Behavioral despair in mice: a primary screening test for antidepressants. Arch. Int. Pharmacodyn. Ther., 1977, 229(2), 327-336.
[PMID: 596982]
[23]
Zhao, J.; Nakamura, N.; Hattori, M.; Kuboyama, T.; Tohda, C.; Komatsu, K. Withanolide derivatives from the roots of Withania somnifera and their neurite outgrowth activities. Chem. Pharm. Bull. (Tokyo), 2002, 50(6), 760-765.
[http://dx.doi.org/10.1248/cpb.50.760] [PMID: 12045329]
[24]
Ahmed, R.F.; Abdel-Rahman, R.F.; Farid, O.A.; El-Marasy, S.A.; Hessin, A.F. Combined hepatoprotective and antidepressant effects of resveratrol in an acute model of depression. Bull. Fac. Pharm. Cairo Univ., 2014, 52(2), 191-197.
[http://dx.doi.org/10.1016/j.bfopcu.2014.06.002]
[25]
Lakshmi, B.V.S.; Sudhakar, M. Screening of Psidium guajava leaf extracts for antistress activity in different experimental animal models. Pharmacogn. Rev., 2009, 1(6), 359.
[26]
Kumar, V.; Singh, P.N.; Jaiswal, A.K.; Bhattacharya, S.K. Antidepressant activity of Indian Hypericum perforatum Linn in rodents. Indian J. Exp. Biol., 1999, 37(12), 1171-1176.
[PMID: 10865882]
[27]
Kim, K.W.; Kim, H.D.; Jung, J.S.; Woo, R.S.; Kim, H.S.; Suh, H.W.; Kim, Y.H.; Song, D.K. Characterization of antidepressant-like effects of p-synephrine stereoisomers. Naunyn Schmiedebergs Arch. Pharmacol., 2001, 364(1), 21-26.
[http://dx.doi.org/10.1007/s002100100416] [PMID: 11485034]


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Article Details

VOLUME: 16
ISSUE: 5
Year: 2020
Published on: 16 July, 2020
Page: [681 - 686]
Pages: 6
DOI: 10.2174/1573407215666190308155305
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