ISOLATION OF CYTOTOXIC CONSTITUENT FROM BIOACTIVITY GUIDED FRACTION OF ALYSICARPUS MONILIFER L. (DC.)
DOI:
https://doi.org/10.22159/ijpps.2019v11i8.33931Keywords:
Alysicarpus monilifer, Betulinic acid, Pentacyclic triterpenoid, Phytochemistry, In vitro cytotoxicity, PhytopharmacologyAbstract
Objective: Alysicarpus monilifer (Family Papilionaceae) has been used in the Indigenous system of medicine in tumor removal. The present study was designed to isolate and identify the constituent responsible for cytotoxic (anti-tumor) effects of the plant Alysicarpus monilifer.
Methods: The plant was powdered and extracted to give a methanolic extract. Initially, Hexane, chloroform, ethyl acetate and methanolic fractions of the methanolic extract of the plant were subjected to cytotoxic screening using cell line based assay (MTT assay and NRU assay). The chloroform fraction showed significant cytotoxicity, so it was further subjected to column chromatography, to separate the cytotoxic phytoconstituent. The cell lines selected were breast cancer cells (MCF-7 and MDA-MB-468) and Liver cancer cells (HepG2 and HLE cell). Results were calculated as percentage growth inhibition with respect to untreated (control) cells versus treated cells.
Result: A triterpene, Betulinic acid, was isolated from the aerial parts of Alysicarpus monilifer. The cytotoxic activity of the identified compound against MCF-7, MDA-MB-231, HLE and HepG2 cells was also found to be highly significant with 90% growth inhibition.
Conclusion: The triterpene was identified to be betulinic acid, to which the cytotoxic activity can be attributed. It is a first report of isolation of betulinic acid from the Alysicarpus species.
Downloads
References
Chavan AR, Oza GM. The flora of pavagadh. Vadodara: Department of Botany, M. S. University of Baroda; 1966.
Shah GL. Flora of gujarat state. 1st Edn. Part І. Vallabh vidyanagar: Sardar Patel University Press; 1978.
Guha Bakshi DN. Flora of Murshidabad district West Bengal, India. Jodhpur: Scientific Publishers; 1984.
Rao RR, Haridasan K. An ethnobotanical survey of medicinal and other useful plants from North-East India. J Econ Tax Bot 1991;15:423-36.
Jain P, Sahu TR. An ethnobotanical study of noradehi sanctuary park of Madhya Pradesh, India: Native plant remedies for scorpion sting and snakebite. J Econ Tax Bot 1993;17:315-28.
Radhakrishnan K, Pandurangan AG, Pushpangadan P, Sasidharan A. Less known ethnomedicinal plants of Kerala state and their conservation. Ethnobotany 1996;8:82-4.
Sankaranarayanan AS. Folklore medicines for jaundice from Coimbatore and palghat districts of Tamil Nadu and Kerala, India. Ancient Sci Life 1988;7:175-9.
Mitchell SA. The jamaican root tonics: a botanical reference. Focus Altern Complement Ther 2011;16:271–80.
Sikarwar RLS, Kaushik JP. Folk medicines of morena district, Madhya Pradesh, India. Int J Pharmacogn 1993;31:283-7.
Singh KK, Prakash A. Indigenous phytotherapy among the gond tribes of Uttar Pradesh, India. Ethnobotany 1994;6:37-41.
Anonymous. Reviews on indian medicinal plants. Vol. 2. New Delhi: ICMR; 2004.
Kumari M, Rao G, Padmaja V. Role of vitexin and isovitexin in hepatoprotective effect of Alysicarpus monilifer linn against CCl4 induced hepatotoxicity. Phytopharmacol 2012;3:273-85.
Riaz N, Anis I, Khan P, Shah R, Malik A. Alysinol-a new triterpene from Alysicarpus monolifer. Nat Prod Lett 2003;16:415-8.
Kakrani PH, Kakrani BH, Kakrani HN. Isolation of stigmasterol and ursolic acid from methanolic extract of aerial parts of Alysicarpus monilifer L. (DC.) (papilionaceae). Pharma Sci Mon 2012;3:2185-91.
Kakrani P, Kakrani H, Saluja A. Pharmacognostic evaluation of aerial parts of Alysicarpus monilifer L. (Dc.). Int J Pharm Pharm Sci 2011(a);3:128-34.
Kakrani P, Kakrani H, Saluja A. Evaluation of anti-inflammatory activity of methanolic extract of the aerial parts of Alysicarpus monilifer L. (DC.). J Pharm Res 2011(b);4:3529-20.
Kakrani P, Kakrani H, Saluja A. Evaluation of analgesic activity of methanolic extract of the aerial parts of Alysicarpus monilifer L.(DC.). Pharma Sci Mon 2011(c):2078-84.
Karthikeyan K. Antidiabetic activities of Alysicarpus monilifer in streptozotocin-induced diabetic rats. Ph. D Thesis. Tamil Nadu: Annamalai University; 2017.
Hossain MA, Ismail Z. Isolation and characterization of triterpenes from the leaves of Orthosiphon stamineus. Arab J Chem 2013;6:295–8.
Harborne J. Phytochemical methods. 2nd Edn. London: Chapan and Hall Ltd; 1973.
Fotakis G, Timbrell A. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett 2006;160:171-7.
Borenfreund E, Babich H, Martin-Alguacil N. Comparisons of two in vitro cytotoxicity assays-the neutral red (NR) and tetrazolium MTT tests. Toxicol In Vitro 1988;2:1-6.
Naik SB, Deshpande SR. Cytotoxicity of stem extracts of selected cassia species against hela and breast cancer cell lines in vitro. Asian J Pharm Clin Res 2017;10:80-2.
Kakrani PH, Kakrani HN, Raval M. Cytotoxic effect of Corchorus depressus against HepG2 and HLE human liver cancer cells. Asian J Pharm Clin Res 2018;11:187-9.
Miller WR. Aromatase inhibitors: mechanism of action and role in the treatment of breast cancer. Semin Oncol 2003;30(4, Suppl 14):3-11.
Levine L. Tamoxifen and the rafoxifene analog LY117018: their effects on arachidonic acid release from cells in culture and on prostaglandin I2 production by rat liver cells. BMC Cancer 2004;13:4-49.
Avendano C, Menendez JC. Anticancer drugs acting via radical species, photosensitizers and photodynamic therapy of cancer. In: Avendano C, Menendez JC. Medicinal Chemistry of Anticancer Drugs. Elsevier; 2008.
Waheeda A, Barbara J, Barton SJ, Khan GM, Najm-Us-Saqib Q, Hussain M, et al. Novel acylated steroidal glycosides from Caralluma tuberculata induced caspase-dependent apoptosis in cancer cells. J Ethnopharmacol 2011;137:1189-96.
Green DR, Kroemer G. The pathophysiology of mitochondrial cell death. Science 2004;305:626-9.
Winckler J. Vital staining of lysosomes and other organelles of the rat with neutral red. Prog Histochem Cytochem 1974;6:1-89.
Chudzik M, Korzonek Szlacheta I, Krol W. Triterpenes as potentially cytotoxic compounds. Molecules 2015;20:1610-25.
Petronelli A, Gaetano P, Ugo T. Triterpenoids as new promising anticancer drugs. Anti Cancer Drugs 2009;20:880-9.
Du JR, Long FY, Chen C. Research progress on natural triterpenoid saponins in the chemoprevention and chemotherapy of cancer. Enzymes 2014;36:95-130.
Wagner H, Bladt S. Plant drug analysis: a thin layer chromatography atlas. 2nd edn. New York: Springer Verlag Berlin Heidelberg; 2011.
Silverstein RM, Webster FX. Spectrometric identification of organic compounds. 6th Edn. New Delhi: Wiley India Pvt. Ltd; 2009.
William Kemp. Organic spectroscopy. Pennsylvania State University: Macmillan; 1991.
Cichewicz RH, Kouzi SA. Chemistry, biological activity, and chemotherapeutic potential of betulinic acid for the prevention and treatment of cancer and HIV infection. Med Res Rev 2004;24:90-114.
Haque ME, Shekhar HU, Mohamad AU, Rahman H, Mydul Islam AKM, Hossain MS. Triterpenoids from the stem bark of Avicennia officinalis. Dhaka University J Pharma Sci 2006;1-2:53-7.
Mukherjee R, Kumar V, Srivastava SK, Agarwal SK, Burman AC. Betulinic acid derivatives as anticancer agents: structure-activity relationship. Anti Cancer Agents Med Chem 2006;6:271-9.
Aslanturk OS. In vitro cytotoxicity and cell viability assays: principles, advantages and disadvantages. In: Larramendy ML, Soloneski S. Genotoxicity-a predictable Risk to our actual world. London: IntechOpen limited; 2017. p. 1-17.
Chang HM, But PPH. Pharmacology and applications of chinese materia medica. Singapore: World Scientific Publishing; 1986.
Kapoor LD. CRC handbook or ayurvedic medicinal plants. Boca Raton, USA: CRC Press; 1990.
Wei L, Brossi A, Susan L, Bastow KF, Lee K. Antitumor agents 248-chemistry and antitumor activity of tylophorine-related alkaloids. Studies Natl Prod Chem Elsevier 2008;34:3–34.
Hartwell JL. Plants used against cancer. Massachusetts: Quarterman Publications; 1982.
Cragg G, Newmann DJ. Biodiversity: a continuing source or novel drug leads. Pure Appl Chem 2005;77:7-24.
Carter Jr WH, Wampler GL, Stabelien DM, Campbell ED. Drug activity and therapeutic synergism in cancer treatment. Cancer Res 1982;42:2963-71.
Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. 2nd Edn. Oxford, UK: Charendon Press; 1988.
Repetto G, Del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc 2008;3:1125-31.
Feng C, Ma W, Song Y, Guo Q, Wu Q, Zheng W. Detection of cell apoptosis by MTT assay. Di Yi Jun Yi Da Xue Xue Bao 2002;22:262-3.