SECONDARY METABOLITES FROM RICE CULTURE OF ASPERGILLUS SP. ISOLATED FROM MELALEUCA SUBULATA (CHEEL) CRAVEN LEAVES AND THEIR ANTICANCER ACTIVITY
Keywords:Anthraquinones, Apoptotic, Aspergillus sp, Melaleuca subulata, Necrotic, Xanthone
Objective: Aspergillus fungus is a rich source of natural products with broad biological activities. This study was conducted to identify secondary metabolites from the rice culture of Aspergillus species isolated from Melaleuca subulata leaves and evaluated their anticancer activity.
Methods: Ethyl acetate extract was fractionated on silica gel and Sephadex columns. Structures of the compounds were established using physical and chemical methods. Cytotoxic activities of the extract and pure compounds against two human cancer cell lines (Mcf-7and Hep G2) were evaluated using microculture tetrazolium assay as well as the mode of the cytotoxicity was evaluated. Molecular docking studies have been performed using the Hsp 90 enzyme as an anticancer target.
Results: Methyl linoleate (1), arugosin C (2), ergosterol (3), sterigmatocystin (4), diorcinol (5), alternariol-5-O-methyl ether (6), averufin (7), averufanin (8), and alternariol (9) were identified from ethyl acetate extract. All tested compounds exhibit week activity against MCF-7 and Hep G2 cell lines but a mixture of compounds 7 and 8 is considered to be more active towards both MCF-7 and Hep G 2 in comparison to other compounds. Compound 4 exhibits moderate activity against Hep G2 only as well as the ethyl acetate extract exerts moderate activity against MCF-7 cell line Moreover, compound 4 and a mixture of 7 and 8 caused a decrease in the number of Hep G2 cancer cells due to apoptotic and necrotic processes. Most active anticancer candidates 7 and 8 showed binding to the active site similar to geldanamycin reference ligand.
Conclusion: Secondary metabolites identified from Aspergillus sp. and their anticancer activity were evaluated. Molecular docking suggested active candidates as Hsp 90 inhibitors.
Chang HS, Lin CH, Chen YS, Wang HC, Chan HY, Hsieh SY, et al. Secondary metabolites of the endophytic fungus Lachnum abnorme from Ardisia cornudentata. Int J Mol Sci 2016;17:1512.
da Silva Ribeiro A, Polonio JC, Costa AT, Dos Santos CM, Rhoden SA, Azevedo JL, et al. Bioprospection of culturable endophytic fungi associated with the ornamental plant Pachystachys lutea. Curr Microbiol 2018;75:588-96.
Kusari S, Singh S, Jayabaskaran C. Rethinking production of taxol(R) (paclitaxel) using endophyte biotechnology. Trends Biotechnol 2014;32:304-11.
Wang XJ, Min CL, Ge M, Zuo RH. An endophytic sanguinarine-producing fungus from Macleaya cordata, Fusarium proliferatum BLH51. Curr Microbiol 2014;68:336-41.
Pan F, Su TJ, Cai SM, Wu W. Fungal endophyte-derived Fritillaria unibracteata var. wabuensis: diversity, antioxidant capacities in vitro and relations to phenolic, flavonoid or saponin compounds. Sci Rep 2017;7:42008.
Frisvad JC, Larsen TO. Chemodiversity in the genus Aspergillus. Appl Microbiol Biotechnol 2015;99:7859-77.
Bladt TT, Frisvad JC, Knudsen PB, Larsen TO. Anticancer and antifungal compounds from Aspergillus, Penicillium, and other filamentous fungi. Molecules 2013;18:11338-76.
Zhang H, Tang Y, Ruan C, Bai X. Bioactive secondary metabolites from the endophytic Aspergillus genus. Rec Nat Prod 2016;10:1-16.
Li ZX, Wang XF, Ren GW, Yuan XL, Deng N, Ji GX, et al. Prenylated diphenyl ethers from the marine algal-derived endophytic fungus Aspergillus tennesseensis. Molecules 2018;23:2368.
Vadlapudi V, Borah N, Yellusani KR, Gade S, Reddy P, Rajamanikyam M, et al. Aspergillus secondary metabolite database, a resource to understand the secondary metabolome of Aspergillus genus. Sci Rep 2017;7:7325.
Innis MA, Gelfand DH, Sninsky JJ, White TJ. PCR protocols: a guide to methods and applications: Academic Press; 2012.
James J, Thomas J. Anticancer activity of microalgae extract on human cancer cell line (mg-63). Asian J Pharm Clin Res 2019;12:139-42.
Stebbins CE, Russo AA, Schneider C, Rosen N, Hartl FU, Pavletich NP. Crystal structure of an Hsp 90–geldanamycin complex: targeting of a protein chaperone by an antitumor agent. Cell 1997;89:239-50.
Allinger NL. Conformational analysis. 130. MM2. A hydrocarbon force field utilizing V1 and V2 torsional terms. J Am Chem Soc 1977;99:8127-34.
Profeta Jr S, Allinger N. Molecular mechanics calculations on aliphatic amines. J Am Chem Soc 1985;107:1907-18.
Diaz MF, Gavin JA. Characterization by NMR of ozonized methyl linoleate. J Brazil Chem Soc 2007;18:513-8.
Shirane N, Takenaka H, Ueda K, Hashimoto Y, Katoh K, Ishii H. Sterol analysis of DMI-resistant and-sensitive strains of Venturia inaequalis. Phytochemistry 1996;41:1301-8.
Martinez M, Alvarez ST, Campi MG, Bravo JA, Vila JL. Ergosterol from the mushroom Laetiporus sp.: isolation and structural characterization. Rev Bol Quim 2015;32:90-4.
Nowak R, Drozd M, Mendyk E, Lemieszek M, Krakowiak O, Kisiel W, et al. A new method for the isolation of ergosterol and peroxyergosterol as active compounds of Hygrophoropsis aurantiaca and in vitro antiproliferative activity of isolated ergosterol peroxide. Molecules 2016;21:946.
Davies J, Kirkaldy D, Roberts JC. 437 Studies in mycological chemistry. Part VII. Sterigmatocystin, a metabolite of Aspergillus versicolor (Vuillemin) tiraboschi. J Chem Soc 1960;0:2169-78.
Ballantine JA, Ferrito V, Hassall CH, Jenkins ML. The biosynthesis of phenols. Part XXIV. Arugosin C, a metabolite of a mutant strain of Aspergillus rugulosus. J Chem Soc Perkin Trans 1973;1:1825-30.
Hawas UW, El-Beih AA, El-Halawany AM. Bioactive anthraquinones from endophytic fungus Aspergillus versicolor isolated from red sea algae. Arch Pharm Res 2012;35:1749-56.
Zhu F, Lin Y. Three xanthones from a marine-derived mangrove endophytic fungus. Chem Nat Compd 2007;43:132-5.
Pachler KG, Steyn PS, Vleggaar R, Wessels PL. Carbon-13 nuclear magnetic resonance assignments and biosynthesis of aflatoxin B1 and sterigmatocystin. J Chem Soc Perkin 1976;1:1182-9.
Itabashi T, Nozawa K, Nakajima S, Kawai KI. A new azaphilone, falconensin H, from Emericella falconensis. Chem Pharm Bull 1993;41:2040-1.
Hassan A. Novel natural products from endophytic fungi of Egyptian medicinal plants-chemical and biological characterization [Dissertation]. Düsseldorf: Universität Düsseldorf; 2007.
De Souza GD, Mithofer A, Daolio C, Schneider B, Rodrigues-Filho E. Identification of Alternaria alternata mycotoxins by LC-SPE-NMR and their cytotoxic effects to soybean (Glycine max) cell suspension culture. Molecules 2013;18:2528-38.
Tan N, Tao Y, Pan J, Wang S, Xu F, She Z, et al. Isolation, structure elucidation, and mutagenicity of four alternariol derivatives produced by the mangrove endophytic fungus No. 2240. Chem Nat Compd 2008;44:296-300.
Dagne E, Yenesew A, Asmellash S, Demissew S, Mavi S. Anthraquinones, pre-anthraquinones, and isoeleutherol in the roots of Aloe species. Phytochemistry 1994;35:401-6.
Chen M, Shao C-L, Kong C-J, She Z-G, Wang C-Y. A new anthraquinone derivative from a gorgonian-derived fungus Aspergillus sp. Chem Nat Compd 2014;50:617-20.
Cole RJ, Cox RH. Handbook of toxic fungal metabolites: Academic Press.; 1981.
Gorst-Allman CP, Pachler KG, Steyn PS, Wessels PL. Carbon-13 nuclear magnetic resonance assignments of some fungal C 20 anthraquinones; their biosynthesis in relation to that of aflatoxin B 1. J Chem Soc Perkin 1 1977:2181-88.
Shao C, Wang C, Wei M, Li S, She Z, Gu Y, et al. Structural and spectral assignments of six anthraquinone derivatives from the mangrove fungus (ZSUH‐36). Magn Reson Chem 2008;46:886-9.
Özkaya FC, Ebrahim W, El-Neketi M, Tanrıkul TT, Kalscheuer R, Müller WE, et al. Induction of new metabolites from the sponge-associated fungus Aspergillus carneus by OSMAC approach. Fitoterapia 2018,131:9-14.
Metha SD, Paliwal S. Phytochemical analysis, liquid chromatography, and mass spectroscopy and in vitro anticancer activity of Annona squamosa seeds lin. Asian J Pharm Clin Res 2018;11:101-3.
Examination IN, Wulandari A, Putihuspa DH, Andayaningsih P. Cytotoxicity of an aromatic compound from an endophytic fungus, Cladosporium sp. En-s01. Int J Curr Pharm Res 2018;10:10-12
McGuire S. World cancer report 2014. Geneva, Switzerland: World Health Organization, the international agency for research on cancer, WHO Press, 2015. In: Oxford University Press; 2016.
Suja M, Vasuki S, Sajitha N. Anticancer activity of compounds isolated from marine endophytic fungus Aspergillus terreus. World J Pharm Pharm Sci 2014;3:661-72.
Zhivotovsky B, Joseph B, Orrenius S. Tumor radiosensitivity, and apoptosis. Exp Cell Res 1999;248:10-7.
Liu Y, Du M, Zhang G. Proapoptotic activity of aflatoxin B1 and sterigmatocystin in HepG2 cells. Toxicol Rep 2014;1:1076-86.
Essigmann J, Barker L, Fowler K, Francisco M, Reinhold V, Wogan G. Sterigmatocystin-DNA interactions: Identification of a major adduct formed after metabolic activation in vitro. Proc Natl Acad Sci 1979;76:179-83.
Akhtar MN, Zareen S, Yeap SK, Ho WY, Lo KM, Hasan A, et al. Total synthesis, cytotoxic effects of damnacanthal, nordamnacanthal and related anthraquinone analogs. Molecules 2013;18:10042-55.
Hsin LW, Wang HP, Kao PH, Lee O, Chen WR, Chen HW, et al. Synthesis, DNA binding, and cytotoxicity of 1, 4-bis (2-amino-ethyl amino) anthraquinone–amino acid conjugates. Bioorg Med Chem 2008;16:1006-14.
Pors K, Paniwnyk Z, Teesdale Spittle P, Plumb JA, Willmore E, Austin CA, et al. Alchemix: a novel alkylating anthraquinone with potent activity against anthracycline-and cisplatin-resistant ovarian cancer. Mol Cancer Ther 2003;2:607-10.