Toxicity Studies on Leaf Extracts of Alternanthera brasiliana (L.) Kuntze and Alternanthera bettzickiana (Regel) Voss

Kasthuri O R; Ramesh B

doi:10.7324/JAPS.2018.81011

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Abstract

Herbal medicine is the source for the search of many novel therapeutic compounds in developing countries. Before used as medicine, drugs from plant origin must be ensured as safe. Objective: The present work focused to study the in vivo and in vitro toxicity effects of hydroethanolic leaf extracts of Alternanthera brasiliana (A. brasiliana) and Alternanthera bettzickiana (A. bettzickiana). Methods: Sub-acute toxicity studies of hydroethanolic leaf extracts of A. brasiliana and A. bettzickiana were carried out in vivo on mice. 11 groups of albino mice were treated with five different doses of the hydroethanolic leaf extracts of A. brasiliana and A. bettzickiana orally for 14 days. General appearance and behavior were observed for 14 consecutive days. Effect on hematological parameters and histopathological changes were also monitored. Cytotoxicity was assessed by observing its toxicity in vitro against DLA cell line by using MTT assay. Results: Sub-acute toxicity studies results showed that up to the tested dose of 2000 mg/kg bwt in both extract treatments, throughout the 14 days of treatment, the extract does not produced any toxicity symptoms. MTT assay indicated that both leaf extracts exhibited significant concentration-dependent in vitro cytotoxic activity against DLA cell line. Hydroethanolic leaf extract of A. bettzickiana exhibited more potent cytotoxic effects on DLA cells than A. brasiliana extract. Conclusion: No toxicity related symptoms were observed in different doses of leaf extract treated mice groups. So the LD50 values of the tested leaf extracts were more than 2000 mg/kg bwt.

Keyword: Cytotoxicity LD50 hematology MTT assay histopathology.

Citation:

Kasthuri OR, Ramesh B. Toxicity Studies on Leaf Extracts of Alternanthera brasiliana (L.) Kuntze and Alternanthera bettzickiana (Regel) Voss. J App Pharm Sci, 2018; 8(10): 082- 089.

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Reference

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Lemkebthomas L, Williams DA, Roche VF, William ZS. Foye's principles of medicinal chemistry., 6th edition, 2008; 1147-8.
Moongkarndi P, Kosem N, Kaslungka S, Luanratana O, Pongpan N, Neungton N. Antiproliferation, antioxidation and induction of apoptosis by Garcinia mangostana (mangosteen) on SKBR3 human breast cancer cell line. J Ethnopharmacol, 2004; 90:161-6.https://doi.org/10.1016/j.jep.2003.09.048
Kinzler KW, Vogelstein B. Introduction. The Genetic Basis of Human Cancer. 2nd ed. New York: McGraw-Hill Publishers; 2002.
Newman DJ, Cragg GM, Snader KM. Natural Products as Sources of New Drugs. J Nat Prod, 2003; 66:1022-103.https://doi.org/10.1021/np030096l
Liu WJ. Introduction to traditional herbal medicines and their study. In: Liu JH, editor. Traditional Herbal Medicine Research Methods. New Jersey: John Wiley & Sons, Inc; 2011; 1–26.https://doi.org/10.1002/9780470921340
Lopes LDC, Albano F, F Laranja F, Alves LM and Silva LFM, et al. Toxicological evaluation by in vitro and in vivo assays of an aqueous extract prepared from Echinodorus macrophyllus leaves. Toxicol Lett, 2000; 116:189-98.https://doi.org/10.1016/S0378-4274(00)00220-4
Saad B, Azaizeh H, Abu-Hijleh G, Said S. Safety of traditional Arab herbal medicine. Evidence-Based Complement Altern Med, 2006; 3:433-9.https://doi.org/10.1093/ecam/nel058
Wojcikowski K, Johnson DW, Gobé G. Medicinal herbal extracts – renal friend or foe? Part one: The toxicities of medicinal herbs. Nephrology (Carlton), 2004; 9:313–8.https://doi.org/10.1111/j.1440-1797.2004.00310.x
Anisuzzaman ASM, Sugimoto N, Sadik G, Gafur MA. Sub-acute toxicity study of 5-Hydroxy-2(Hydroxy-Methyl) 4H-pyran-4 One, isolated from Aspergillus fumigatus. Pak J Biol Sci, 2001; 4:1012-5.https://doi.org/10.3923/pjbs.2001.1012.1015
Rang HP, Dale M, Ritter. Pharmacology. Volume 13. 4th Ed. New York, NY, USA; Churchill Livingstone, 2001.
Hussien M Alwadie. Morphology and Distribution of Three Genera of Amaranthaceae in the South Western Area of Saudi Arabia. J King Saud Univ, 2005; 18(1):51-62.
Kumar S, Singh P, Mishra G, Srivasta S, Jha KK, Khosa RL. Phytopharmacological review of Alternanthera brasiliana (Amaranthaceae). Asian J Plant Sci Res, 2011; 1:41–7.
Suphanthip P, Waranya C, Orawan M, Pradit P, Kanokwan J. Alternanthera sessilis and Alternanthera bettzickiana Improved Superoxide Dismutase and Catalase Activities in the Livers of Ovariectomized Mice. Journal of Applied Biopharmaceutics and Pharmacokinetic, 2013; 1(2):64- 71.
Kasthuri OR and Ramesh B. Phytochemical screening and in vitro antioxidant activities of leaf extracts of Alternanthera brasiliana (L.). Kuntze and Alternanthera bettzickiana Regel. Asian J Pharm Clin Res, 2018; 11(6):266-272. Jonsson M, Jestoi M, Nathanail AV, Kokkonen UM, Anttila M, Koivisto P, Karhunen P, Peltonen K. Application of OECD Guideline 423 in assessing the acute oral toxicity of moniliformin. Food Chem Toxicol, 2013; 53:27–32.https://doi.org/10.1016/j.fct.2012.11.023
Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 1983; 65:55-63.https://doi.org/10.1016/0022-1759(83)90303-4
Wilson AP, John RW. Animal cell culture: a practical approach. oxford: Oxford University Press, 2000.
Yan-Wei H, Chun-Yu L, Chong-Min D, Wen-Qian W, Zhen-Lun G. Induction of apoptosis in human hepatocarcinoma SMMC-7721 cells in vitro by flavonoids from Astragalus complanatus. J Ethnopharmacol, 2009; 123:293–301.https://doi.org/10.1016/j.jep.2009.03.016
Gesler WM. Therapeutic landscapes: Medical issues in light of the new cultural geography. Soc Sci Med, 1992; 34:735-46.https://doi.org/10.1016/0277-9536(92)90360-3
Bent S, Ko R. Commonly used herbal medicine in the United States: A review. Am J Med, 2004; 116:478-85.https://doi.org/10.1016/j.amjmed.2003.10.036
Hunter P. A toxic brew we cannot live without. EMBO Rep, 2008; 9(1):15–8.https://doi.org/10.1038/sj.embor.7401148
Schulz V, Hansel R, Tyler VE. Rational Phytotherapy: A Physician's Guide to Herbal Medicine; Psychology Press: London, UK, 2001.https://doi.org/10.1007/978-3-642-98093-0
Akhila JS, Deepa S, Alwar MC. Acute toxicity studies and determination of median lethal dose. Current Sci, 2007; 93:917–20.
Parra AL, Yhebra RS, Sardinas IG, Buela LI. Comparative study of the assay of Artemia salina L. and the estimate of the medium lethal dose (LD50 value) in mice, to determine oral acute toxicity of plant extracts. Phytomedicine, 2001; 8:395-400.https://doi.org/10.1078/0944-7113-00044
Rosenthal N, Brown S. The mouse ascending: perspectives for human-disease models. Nat Cell Biol, 2007; 9:993-9.https://doi.org/10.1038/ncb437
Ahmad M, Lim CP, Akowuah GA, Ismail NN, Hashim MA, Hor SY, Ang LF and Yam MF. Safety assessment of standardised methanol extract of Cinnamomum burmannii. Phytomedicine, 2013; 15:1124-30.https://doi.org/10.1016/j.phymed.2013.05.005
Subramanion LJ, Zakaria Z, Chen Y, Lau YL, Latha LY, Sasidharan S. Acute oral toxicity of methanolic seed extract of cassia fistula in mice. Molecules, 2011; 16(6):5268–82.https://doi.org/10.3390/molecules16065268
Walum E. Acute oral toxicity. Environ Health Perspect 1998; 106:497-503.
Klaassen CD. Casarett and Doulls. Toxicology: The Basic Science of Poison. 6th Edn., The McGraw-Hill Companies Inc., New York. 2001.
Nair R, Shukla V, Chanda S. Effect of single dose administration of Polyalthia longifolia (Sonn.) Thw. Var. pendula leaf on gross behavioral assessment in mice. Indian Drugs, 2009; 46:116-23.
Jothy SL, Zuraini Z, Sasidharan S. Phytochemicals screening, DPPH free radical scavenging and xanthine oxidase inhibitiory activities of Cassia fistula seeds extract. J Med Plants Res, 2011; 5(10):1941–7.
Yamthe L, Fokou P, Mbouna C, Keumoe R, Ndjakou B, Djouonzo P, Boyom F. Extracts from Annona Muricata L. and Annona Reticulata L. (Annonaceae) potently and selectively inhibit Plasmodium falciparum. Medicines, 2015; 2(2):55-66.https://doi.org/10.3390/medicines2020055
Schalm OW, Jain NC, Carrol EJ. Veterinary Haematology. 3rd Edn., Lea and Febiger Publication, Philadelphia,1975; 807.
Ajagbonna OP, Onifade KI, Suleiman U. Haematological and biochemical changes in rats given extract of Calotropis procera Sokoto. J Vet Sci, 1999; 1:36-42.
Bosco AD, Gerencser Z, Szendro Z, Ugnai C, Cullere M, et al. Dietary supplementation of spirulina (Arthrospira platensis) and Ðyme Ðymus vulgaris) on rabbit meat appearance, oxidative stability and fatty acid profile during retail display. Meat Sci, 2014; 96:114-9.https://doi.org/10.1016/j.meatsci.2013.06.021
Raza M, Al-Shabanah OA, El-Hadiyah TM, Al-Majed AA. Effect of prolonged vigabatrin treatment on haematological and biochemical parameters in plasma, liver and kidney of Swiss albino mice. Pharmace Scien, 2002; 70:135-45.
Olson H, Betton G, Robinson D, et al. Concordance of the toxicity of pharmaceuticals in humans and in animals. Regulatory Toxicology and Pharmacology, 2000; 32(1):56–67.https://doi.org/10.1006/rtph.2000.1399
Hoffbrand AV, Pettit JE. Essential of Heamatology 3rd edition, Blackwell Sci Inc., USA, 1997.
Jain N, Sharma P, Sharma N, Joshi SC. Haemato-biochemical profile following sub acute toxicity of malathion in male albino rats. Avicenna J Phytomed, 2009; 2:500–506.
Ojezele, Matthew Obaineh, Agunbiade, Shadrach. Phytochemical constituents and medicinal properties of different extracts of Anacardium occidentale and Psidium Guajava. Asian Journal of Biomedical and Pharmaceutical Sciences, 2013; 3(16):20-23.
Purves WK, David S, Gordon HO, Craig HH. Life: The Science of Biology. 7th edition, Mass: Sinauer Associates, Sunderland, 2004; pp: 954.
Odeyemi OO, Yakubu MT, Masika PJ, Afolayan AJ. Toxicological evaluation of the essential oil from Menthalongifolia L. subsp. capensis leaves in rats. J Med Food, 2009; 12:669–74.https://doi.org/10.1089/jmf.2008.0136
Tousson E, El-Moghazy M, El-Atrsh E. The possible effect of diets containing Nigella sativa and Thymus vulgarison blood parameters and some organs structure in rabbit. Toxicol Ind Health, 2011; 27(2);107- 16.https://doi.org/10.1177/0748233710381891
Weingand K, Brown G, Hall R, Davies D, Gossett K, et al. Harmonization of animal clinical pathology testing in toxicity and safety studies. The Joint Scientific Committee for International Harmonization of Clinical Pathology Testing. Fundam Appl Toxicol, 1996; 29(2):198-201.https://doi.org/10.1006/faat.1996.0022
Ajibade TO, Olayemi FO, Arowolo ROA. The haematological and biochemical effects of methanol extract of the seeds of Moringa oleifera in rats. J Medic Plants Res, 2012; 6(2):615-21.https://doi.org/10.5897/JMPR11.1258
Bretagnol F, et al. The effect of sterilization processes on the bioadhesive properties and surface chemistry of a plasma-polymerized polyethylene glycol film: XPS characterization and L929 cell proliferation tests. Acta Biomater, 2008; 4:1745-51.https://doi.org/10.1016/j.actbio.2008.06.013
Cardellina JH, Fuller RW, Gamble WR, Westergaard C, Boswell J, Munro MHG, Currens M, Boyd MP. Evolving strategies for the selection dereplication and prioritization of antitumor and HIV inhibitory natural products extracts. In: Bohlin, L., Bruhn, J.G. (Eds), Bioassay Methods in Natural Product Research and Development. Kluwer Academic Publishers, Dordrecht 1999; pp. 25-36.https://doi.org/10.1007/978-94-011-4810-8_3
Butlet MS. The role of natural product chemistry in drug discovery. J Nat Products, 2004; 67:2141-2153.https://doi.org/10.1021/np040106y
Shoeb M. Anticancer agents from medicinal plants. Bang J Pharmacol, 2006; 1:35-41.
Babu BH, Shylesh BS, and Padikkala J. Tumour reducing and anticarcinogenic aactivity of Acanthus ilicifolius in mice. J Ethno Pharmacol, 2002; 79:27–33.https://doi.org/10.1016/S0378-8741(01)00347-6

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Kaufman D, Chabner BA. Clinical strategies for cancer treatment: The role of drugs. In: Chabner BA, Longo DL, editors. Cancer Chemotherapy and Biotherapy: Principles and Practice. Philadelphia: Lippincott-Raven 1996; 1-16.

Lemkebthomas L, Williams DA, Roche VF, William ZS. Foye's principles of medicinal chemistry., 6th edition, 2008; 1147-8.

Moongkarndi P, Kosem N, Kaslungka S, Luanratana O, Pongpan N, Neungton N. Antiproliferation, antioxidation and induction of apoptosis by Garcinia mangostana (mangosteen) on SKBR3 human breast cancer cell line. J Ethnopharmacol, 2004; 90:161-6.https://doi.org/10.1016/j.jep.2003.09.048

Kinzler KW, Vogelstein B. Introduction. The Genetic Basis of Human Cancer. 2nd ed. New York: McGraw-Hill Publishers; 2002.

Newman DJ, Cragg GM, Snader KM. Natural Products as Sources of New Drugs. J Nat Prod, 2003; 66:1022-103.https://doi.org/10.1021/np030096l

Liu WJ. Introduction to traditional herbal medicines and their study. In: Liu JH, editor. Traditional Herbal Medicine Research Methods. New Jersey: John Wiley & Sons, Inc; 2011; 1–26.https://doi.org/10.1002/9780470921340

Lopes LDC, Albano F, F Laranja F, Alves LM and Silva LFM, et al. Toxicological evaluation by in vitro and in vivo assays of an aqueous extract prepared from Echinodorus macrophyllus leaves. Toxicol Lett, 2000; 116:189-98.https://doi.org/10.1016/S0378-4274(00)00220-4

Saad B, Azaizeh H, Abu-Hijleh G, Said S. Safety of traditional Arab herbal medicine. Evidence-Based Complement Altern Med, 2006; 3:433-9.https://doi.org/10.1093/ecam/nel058

Wojcikowski K, Johnson DW, Gobé G. Medicinal herbal extracts – renal friend or foe? Part one: The toxicities of medicinal herbs. Nephrology (Carlton), 2004; 9:313–8.https://doi.org/10.1111/j.1440-1797.2004.00310.x

Anisuzzaman ASM, Sugimoto N, Sadik G, Gafur MA. Sub-acute toxicity study of 5-Hydroxy-2(Hydroxy-Methyl) 4H-pyran-4 One, isolated from Aspergillus fumigatus. Pak J Biol Sci, 2001; 4:1012-5.https://doi.org/10.3923/pjbs.2001.1012.1015

Rang HP, Dale M, Ritter. Pharmacology. Volume 13. 4th Ed. New York, NY, USA; Churchill Livingstone, 2001.

Hussien M Alwadie. Morphology and Distribution of Three Genera of Amaranthaceae in the South Western Area of Saudi Arabia. J King Saud Univ, 2005; 18(1):51-62.

Kumar S, Singh P, Mishra G, Srivasta S, Jha KK, Khosa RL. Phytopharmacological review of Alternanthera brasiliana (Amaranthaceae). Asian J Plant Sci Res, 2011; 1:41–7.

Suphanthip P, Waranya C, Orawan M, Pradit P, Kanokwan J. Alternanthera sessilis and Alternanthera bettzickiana Improved Superoxide Dismutase and Catalase Activities in the Livers of Ovariectomized Mice. Journal of Applied Biopharmaceutics and Pharmacokinetic, 2013; 1(2):64- 71.

Kasthuri OR and Ramesh B. Phytochemical screening and in vitro antioxidant activities of leaf extracts of Alternanthera brasiliana (L.). Kuntze and Alternanthera bettzickiana Regel. Asian J Pharm Clin Res, 2018; 11(6):266-272. Jonsson M, Jestoi M, Nathanail AV, Kokkonen UM, Anttila M, Koivisto P, Karhunen P, Peltonen K. Application of OECD Guideline 423 in assessing the acute oral toxicity of moniliformin. Food Chem Toxicol, 2013; 53:27–32.https://doi.org/10.1016/j.fct.2012.11.023

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 1983; 65:55-63.https://doi.org/10.1016/0022-1759(83)90303-4

Wilson AP, John RW. Animal cell culture: a practical approach. oxford: Oxford University Press, 2000.

Yan-Wei H, Chun-Yu L, Chong-Min D, Wen-Qian W, Zhen-Lun G. Induction of apoptosis in human hepatocarcinoma SMMC-7721 cells in vitro by flavonoids from Astragalus complanatus. J Ethnopharmacol, 2009; 123:293–301.https://doi.org/10.1016/j.jep.2009.03.016

Gesler WM. Therapeutic landscapes: Medical issues in light of the new cultural geography. Soc Sci Med, 1992; 34:735-46.https://doi.org/10.1016/0277-9536(92)90360-3

Bent S, Ko R. Commonly used herbal medicine in the United States: A review. Am J Med, 2004; 116:478-85.https://doi.org/10.1016/j.amjmed.2003.10.036

Hunter P. A toxic brew we cannot live without. EMBO Rep, 2008; 9(1):15–8.https://doi.org/10.1038/sj.embor.7401148

Schulz V, Hansel R, Tyler VE. Rational Phytotherapy: A Physician's Guide to Herbal Medicine; Psychology Press: London, UK, 2001.https://doi.org/10.1007/978-3-642-98093-0

Akhila JS, Deepa S, Alwar MC. Acute toxicity studies and determination of median lethal dose. Current Sci, 2007; 93:917–20.

Parra AL, Yhebra RS, Sardinas IG, Buela LI. Comparative study of the assay of Artemia salina L. and the estimate of the medium lethal dose (LD50 value) in mice, to determine oral acute toxicity of plant extracts. Phytomedicine, 2001; 8:395-400.https://doi.org/10.1078/0944-7113-00044

Rosenthal N, Brown S. The mouse ascending: perspectives for human-disease models. Nat Cell Biol, 2007; 9:993-9.https://doi.org/10.1038/ncb437

Ahmad M, Lim CP, Akowuah GA, Ismail NN, Hashim MA, Hor SY, Ang LF and Yam MF. Safety assessment of standardised methanol extract of Cinnamomum burmannii. Phytomedicine, 2013; 15:1124-30.https://doi.org/10.1016/j.phymed.2013.05.005

Subramanion LJ, Zakaria Z, Chen Y, Lau YL, Latha LY, Sasidharan S. Acute oral toxicity of methanolic seed extract of cassia fistula in mice. Molecules, 2011; 16(6):5268–82.https://doi.org/10.3390/molecules16065268

Walum E. Acute oral toxicity. Environ Health Perspect 1998; 106:497-503.

Klaassen CD. Casarett and Doulls. Toxicology: The Basic Science of Poison. 6th Edn., The McGraw-Hill Companies Inc., New York. 2001.

Nair R, Shukla V, Chanda S. Effect of single dose administration of Polyalthia longifolia (Sonn.) Thw. Var. pendula leaf on gross behavioral assessment in mice. Indian Drugs, 2009; 46:116-23.

Jothy SL, Zuraini Z, Sasidharan S. Phytochemicals screening, DPPH free radical scavenging and xanthine oxidase inhibitiory activities of Cassia fistula seeds extract. J Med Plants Res, 2011; 5(10):1941–7.

Yamthe L, Fokou P, Mbouna C, Keumoe R, Ndjakou B, Djouonzo P, Boyom F. Extracts from Annona Muricata L. and Annona Reticulata L. (Annonaceae) potently and selectively inhibit Plasmodium falciparum. Medicines, 2015; 2(2):55-66.https://doi.org/10.3390/medicines2020055

Schalm OW, Jain NC, Carrol EJ. Veterinary Haematology. 3rd Edn., Lea and Febiger Publication, Philadelphia,1975; 807.

Ajagbonna OP, Onifade KI, Suleiman U. Haematological and biochemical changes in rats given extract of Calotropis procera Sokoto. J Vet Sci, 1999; 1:36-42.

Bosco AD, Gerencser Z, Szendro Z, Ugnai C, Cullere M, et al. Dietary supplementation of spirulina (Arthrospira platensis) and Ðyme Ðymus vulgaris) on rabbit meat appearance, oxidative stability and fatty acid profile during retail display. Meat Sci, 2014; 96:114-9.https://doi.org/10.1016/j.meatsci.2013.06.021

Raza M, Al-Shabanah OA, El-Hadiyah TM, Al-Majed AA. Effect of prolonged vigabatrin treatment on haematological and biochemical parameters in plasma, liver and kidney of Swiss albino mice. Pharmace Scien, 2002; 70:135-45.

Olson H, Betton G, Robinson D, et al. Concordance of the toxicity of pharmaceuticals in humans and in animals. Regulatory Toxicology and Pharmacology, 2000; 32(1):56–67.https://doi.org/10.1006/rtph.2000.1399

Hoffbrand AV, Pettit JE. Essential of Heamatology 3rd edition, Blackwell Sci Inc., USA, 1997.

Jain N, Sharma P, Sharma N, Joshi SC. Haemato-biochemical profile following sub acute toxicity of malathion in male albino rats. Avicenna J Phytomed, 2009; 2:500–506.

Ojezele, Matthew Obaineh, Agunbiade, Shadrach. Phytochemical constituents and medicinal properties of different extracts of Anacardium occidentale and Psidium Guajava. Asian Journal of Biomedical and Pharmaceutical Sciences, 2013; 3(16):20-23.

Purves WK, David S, Gordon HO, Craig HH. Life: The Science of Biology. 7th edition, Mass: Sinauer Associates, Sunderland, 2004; pp: 954.

Odeyemi OO, Yakubu MT, Masika PJ, Afolayan AJ. Toxicological evaluation of the essential oil from Menthalongifolia L. subsp. capensis leaves in rats. J Med Food, 2009; 12:669–74.https://doi.org/10.1089/jmf.2008.0136

Tousson E, El-Moghazy M, El-Atrsh E. The possible effect of diets containing Nigella sativa and Thymus vulgarison blood parameters and some organs structure in rabbit. Toxicol Ind Health, 2011; 27(2);107- 16.https://doi.org/10.1177/0748233710381891

Weingand K, Brown G, Hall R, Davies D, Gossett K, et al. Harmonization of animal clinical pathology testing in toxicity and safety studies. The Joint Scientific Committee for International Harmonization of Clinical Pathology Testing. Fundam Appl Toxicol, 1996; 29(2):198-201.https://doi.org/10.1006/faat.1996.0022

Ajibade TO, Olayemi FO, Arowolo ROA. The haematological and biochemical effects of methanol extract of the seeds of Moringa oleifera in rats. J Medic Plants Res, 2012; 6(2):615-21.https://doi.org/10.5897/JMPR11.1258

Bretagnol F, et al. The effect of sterilization processes on the bioadhesive properties and surface chemistry of a plasma-polymerized polyethylene glycol film: XPS characterization and L929 cell proliferation tests. Acta Biomater, 2008; 4:1745-51.https://doi.org/10.1016/j.actbio.2008.06.013

Cardellina JH, Fuller RW, Gamble WR, Westergaard C, Boswell J, Munro MHG, Currens M, Boyd MP. Evolving strategies for the selection dereplication and prioritization of antitumor and HIV inhibitory natural products extracts. In: Bohlin, L., Bruhn, J.G. (Eds), Bioassay Methods in Natural Product Research and Development. Kluwer Academic Publishers, Dordrecht 1999; pp. 25-36.https://doi.org/10.1007/978-94-011-4810-8_3

Butlet MS. The role of natural product chemistry in drug discovery. J Nat Products, 2004; 67:2141-2153.https://doi.org/10.1021/np040106y

Shoeb M. Anticancer agents from medicinal plants. Bang J Pharmacol, 2006; 1:35-41.

Babu BH, Shylesh BS, and Padikkala J. Tumour reducing and anticarcinogenic aactivity of Acanthus ilicifolius in mice. J Ethno Pharmacol, 2002; 79:27–33.https://doi.org/10.1016/S0378-8741(01)00347-6