Isolation and Characterization of Antimycobacterial Compounds from Fruits of Aegle marmelos (L.) Correa

Suja K P, Leny Jose, Divya Lakshmanan, Anu G Vidya, Reshmi Nair RJ, R Ajay Kumar

Abstract


Root extracts of Aegle marmelos, an evergreen tree indigenous to India, is an important ingredient in Ayurvedic preparations used for the treatment of pulmonary tuberculosis. Leaves of the tree are used as a remedy for respiratory ailments in folk medicine. The fruit is extensively used in the treatment of various bacterial diseases such as dysentery, diarrhea, etc. The present study was designed to screen organic solvent extracts of fruits of A. Marmelos for their inhibitory activity on Mycobacterium tuberculosis (M. tuberculosis), and to isolate and identify the compounds responsible for antimycobacterial activity. The minimum inhibitory concentration (MIC) of the extract and purified  molecules on virulent laboratory strain M. tuberculosis H37Rv was determined by resazurin microtiter assay (REMA). Structural elucidation of the compounds was carried out employing NMR. Cytotoxicity of the compounds was evaluated against human monocyte THP1-derived macrophages by MTT assay. The hexane extract of A. marmelos fruits inhibited the growth of M. tuberculosis at an MIC of 50 μg/mL. Fractionation of the hexane extract by column chromatography led to the isolation of imperatorin, β-sitosterol, plumbagin, marmesin, marmin,
and stigmasterol which individually inhibited the growth of M. tuberculosis. In vitro cytotoxicity assay revealed that the compounds are not cytotoxic at the concentrations that inhibited the growth of M. tuberculosis.


Keywords


Imperatorin, β-sitosterol, Plumbagin, Marmesin, Marmin, Stigmasterol

Full Text:

PDF

References


Zumla A, Atun R, Maeurer M et al. Viewpoint:

Scientific dogmas, paradoxes and mysteries of latent

Mycobacterium tuberculosis infection. Tropical

Medicine & International Health 2011; 16(1): 79-83.

World Health Organization, Global tuberculosis report,

Geneva: World Health Organization 2016. http://

www.int/tb/publication/global.

Getahun H, Gunneberg C, Granich R et al. HIV

infection-associated tuberculosis: The epidemiology

and the response. Clinical Infectious Diseases, 2010;

(Supplement 3), S201-S207.

Velayati AA, Farnia P, Masjedi MR. Letter to Editor: The

totally drug resistant tuberculosis (TDR-TB). Int J Clin

Exp Med 2013; 6(4): 307-09.

Nguta JM, Appiah-Opong R, Nyarko AK et al. Current

perspectives in drug discovery against tuberculosis

from natural products. International Journal of

Mycobacteriology 2015; 4(3): 165-83.

Gautam R, Saklani A, Jachak SM. Indian medicinal

plants as a source of antimycobacterial agents. Journal

of Ethnopharmacology 2007; 110(2): 200-34.

Lakshmanan D, Werngren J, Jose L et al. Ethyl

p-methoxycinnamate isolated from a traditional antituberculosis

medicinal herb inhibits drug resistant

strains of Mycobacterium tuberculosis in vitro.

Fitoterapia 2011; 82(5): 757-61.

Rahman S, Parvin R. Therapeutic potential of Aegle

marmelos (L.) – An overview. Asian Pacific Journal of

Tropical Disease 2014; 4(1): 71-77.

Baliga MS, Bhat HP, Joseph N et al. Phytochemistry

and medicinal uses of the bael fruit (Aegle marmelos

Correa): A concise review. Food Research International,

; 44(7): 1768-75.

Sharma DR, Sharma DS. Sahasrayogam, re-edited.

Delhi: Chowkhambha Sanskrit Pratisthan 2004.

Kaur R, Kaur H. Antitubercular activity and

phytochemical screening of selected medicinal plants.

Orient J Chem 2015; 31(1).

Ramachandran J. Herbs of Siddha medicine/The first

D book on herbs. Murugan PPatthipagam, Chennai,

India 2008; 156.

Palomino JC, Martin A, Camacho M et al.

Resazurinmicrotiter assay plate: Simple and

inexpensive method for detection of drug resistance

in Mycobacterium tuberculosis. Antimicrobial Agents

and Chemotherapy 2002; 46(8): 2720-22.

Van de Loosdrecht AA, Beelen RHJ, Ossenkoppele GJ

et al. A tetrazolium-based colorimetric MTT assay to

quantitate human monocyte mediated cytotoxicity

against leukemic cells from cell lines and patients with

acute myeloid leukemia. Journal of immunological

methods 1994; 174(1-2): 311-20.

Ghosh K. A furocoumarin, Imperatorin isolated from

Urenalobata L. (Malvaceae). Molbank 2004; 2004(1),

M382.

Chaturvedula VSP, Prakash I. Isolation of Stigmasterol

and β-Sitosterol from the dichloromethane extract

of Rubus suavissimus. International Current

Pharmaceutical Journal 2012; 1(9): 239-42.

Raj G, Kurup R, Hussain AA. Distribution of

naphthoquinones, plumbagin, droserone, and

-O-methyl droserone in chitin-induced and uninduced

Nepenthes khasiana: molecular events in prey capture.Journal of Experimental Botany 2011; 62(15): 5429-36.

Chatterjee MA, Dutta CP, Bhattacharyya MS et al. The

structure of marmin. Tetrahedron Letters 1967; 8(5):

-73.

Jain M, Trivedi A, Mishra SH. TLC determination of

marmesin, a biologically active marker from Feronia

Limonia L. American Journal of Plant Sciences 2010;

(01): 12.

Joshi PV, Patil RH, Maheshwari VL. In vitro antidiarrhoeal

activity and toxicity profile of Aeglemarmelos Correa

ex Roxb. dried fruit pulp. Natural Product Radiance

; 8(5): 498-502.

Chopra RN, Chopra IC. Indigenous drugs of India.

Academic Publishers 1933.

Tawde KV, Gacche RN, Pund MM. Evaluation of

selected Indian traditional folk medicinal plants against

Mycobacterium tuberculosis with antioxidant and

cytotoxicity study. Asian Pacific Journal of Tropical

Disease 2012; 2 (Supplement 2): S685-S691.

Hazra B, Sarkar R, Bhattacharyya S et al. Synthesis of

plumbagin derivatives and their inhibitory activities

against Ehrlich ascites carcinoma in vivo and Leishmania

donovani promastigotes in vitro. Phytotherapy Research

; 16(2): 133-37.

Srinivas P, Gopinath G, Banerji A et al. Plumbagin

induces reactive oxygen species, which mediate

apoptosis in human cervical cancer cells. Molecular

Carcinogenesis 2004; 40(4): 201-11.

Checker R, Sharma D, Sandur SK et al. Anti-inflammatory

effects of plumbagin are mediated by inhibition of

NF-kappaB activation in lymphocytes. International

immunopharmacology 2009; 9(7): 949-58.

Paiva SRD, Figueiredo MR, Aragão TV et al. Antimicrobial

activity in vitro of plumbagin isolated from Plumbago

species. Memorias do Instituto Oswaldo Cruz 2003;

(7): 959-61.

Mossa JS, El‐Feraly FS, Muhammad I. Antimycobacterial

constituents from Juniperus procera, Ferula communis

and Plumbago zeylanica and their in vitro synergistic

activity with isonicotinic acid hydrazide. Phytotherapy

Research 2004; 18(11): 934-37.

Rosselli S, Maggio A, Bellone G et al. Antibacterial and

anticoagulant activities of coumarins isolated from

the flowers of Magydaris tomentosa. Planta medica

; 73(02): 116-20.

Fouotsa H, Mbaveng AT, Mbazoa CD et al. Antibacterial

constituents of three Cameroonian medicinal plants:

Garcinia nobilis, Oricia suaveolens and Balsamocitrus

camerunensis. BMC Complementary and Alternative

Medicine 2013; 13(1): 81.

Rahman A, Na M, Kang SC. Antilisterial potential of

imperatorin and limonin from Poncirus trifoliata Rafin.

Journal of Food Biochemistry 2012; 36(2): 217-23.

Sancho R, Márquez N, Gómez-Gonzalo M et al.

Imperatorin inhibits HIV-1 replication through an Sp1-

dependent pathway. Journal of Biological Chemistry

; 279(36): 37349-59.

Ulazka B, Głowniak K, Sieniawska E. Imperatorinpharmacological

effects and possible implication in

pharmacotherapy. Curr Issues Pharm Med Sci 2012;

(1): 80-87.

Yang LL, Wang MC, Chen LG. Cytotoxic activity of

coumarins from the fruits of Cnidium monnieri on

leukemia cell lines. Planta medica 2003; 69(12): 1091-

Schinkovitz A, Gibbons S, Stavri M et al. Ostruthin:

an antimycobacterial coumarin from the roots of

Peucedanum ostruthium. Planta medica 2003; 69(04):

-71.

Nugroho AE, Riyanto S, Sukari MA et al. Anti-allergic

effects of Marmin, a coumarine isolated from Aegle

marmelos Correa: In vitro study. International Journal

of Phytomedicine 2011; 3(1): 84.

Edilu A, Adane L, Woyessa D. In vitro antibacterial

activities of compounds isolated from roots of

Cayluseaabyssinica. Annals of Clinical Microbiology

and Antimicrobials 2015; 14(1): 15.

Nyila MA, Leonard CM, Hussein AA et al. Activity

of South African medicinal plants against Listeria

monocytogenes biofilms, and isolation of active

compounds from Acacia karroo. South African Journal

of Botany 2015; 78: 220-27.

Navarro-García VM, Luna-Herrera J, Rojas-Bribiesca

MG et al. Antibacterial activity of Aristolochia

brevipes against multidrug-resistant Mycobacterium

tuberculosis. Molecules 2011; 16(9): 7357-64.

Chinchansure AA SNH, Arkile M, Sarkar D et al.

Antimycobacterium activity of coumarins from pulp

of AegleMarmelos (L.) Correa. International Journal

of Basic and Applied Chemical Sciences 2015; 5 (3)

(July-Sep): 39-44.


Refbacks

  • There are currently no refbacks.


Copyright (c) 2018 Journal of Communicable Diseases (P-ISSN: 0019-5138)

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.