CYTOTOXIC LABDANE DITERPENOIDS FROM ANDROGRAPHIS PANICULATA (BURM.F.) NEES

Maria Carmen s Tan; Glenn G Oyong; Chien-Chang Shen; Consolacion Y Ragasa

doi:10.22159/ajpcr.2017.v10i12.19194

Authors

Maria Carmen s Tan Department of Chemistry, De La Salle University, 2401 Taft Avenue, Manila 1004, Philippines.
Glenn G Oyong Department of Biology, De La Salle University, 2401 Taft Avenue, Manila 1004, Philippines.
Chien-Chang Shen National Research Institute of ChineseMedicine, Ministry of Health and Welfare, 155-1, Li-Nong St., Section 2, Taipei, Taiwan.
Consolacion Y Ragasa Department of Chemistry, De La Salle University Science and Technology Complex Leandro V. Locsin Campus, BiÃ±an City, Laguna 4024, Philippines.

DOI:

https://doi.org/10.22159/ajpcr.2017.v10i12.19194

Keywords:

Andrographis paniculata, Andrographolide, 14-deoxyandrographolide, Neoandrographolide, Cytotoxicity

Abstract

Objective: The primary objective of this study was to probe the cytotoxic capacity of the labdane diterpenoids andrographolide (1), 14-deoxyandrographolide (2), 14-deoxy-12-hydroxyandrographolide (3), and neoandrographolide (4) on mutant and wild-type immortalized cell lines.

Methods: Breast adenocarcinoma (MCF-7), colon carcinomas (HCT-116 and HT-29), small cell lung carcinoma (H69PR), human acute monocytic leukemia (THP-1), and wild-type primary normal human dermal fibroblasts - neonatal cells (HDFn) were incubated with 1-4, and the degree of cytotoxicity was analyzed by employing the in vitro PrestoBlueÂ® cell viability assay. Working solutions of 1-4 were prepared in complete cell culture medium to a final non-toxic dimethyl sulfoxide concentration of 0.2%. The plates were incubated at 37Â°C with 5% CO2 in a 98% humidified incubator throughout the assay. Nonlinear regression and statistical analyses were done to extrapolate the half maximal inhibitory concentration 50% (IC50). One-way ANOVA (p<0.05) and multiple comparison, Tukey's post hoc test (p<0.05), were used to compare different pairs of data sets. Results were considered statistically significant at p<0.05.

Results: The highest cytotoxicity index was exhibited by the H69PR and 1 trials which displayed the lowest IC50 value of 3.66 Î¼g/mL, followed by HT-29 treated with 2, HCT-116 and 1 trials, and H69PR treated with 4 (IC50=3.81, 3.82, and 4.19 Î¼g/mL, respectively). Only 1 and 4 were detrimental toward MCF-7, while 1, 3, and 4 were degenerative against H69PR. Tukey's post hoc multiple comparison indicated no significant difference in the cytotoxicity of 1-4 on HCT-116 cells which afforded IC50 values ranging from 3.82 to 5.12 Î¼g/mL. Evaluation of the two colon carcinoma cell lines showed that HCT-116 was categorically more susceptible to cellular damage caused by treatments with 1-4 than was HT-29. Cytotoxicity was not detected in THP-1 and HDFn cells (IC50>100 Î¼g/mL).

Conclusion: Diterpenoids 1-4 isolated from the dichloromethane extract of the leaves of A. paniculata exhibited different cytotoxic activities against MCF-7, HCT-116, HT-29, and H69PR. All constituents had comparable action on HCT-116 cells but were not found to be cytotoxic to normal HDFn cells and mutant THP-1 cells.

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Author Biography

Maria Carmen s Tan, Department of Chemistry, De La Salle University, 2401 Taft Avenue, Manila 1004, Philippines.

Chemistry Department, De La Salle University

Full Professor 10

References

Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011;61(2):69-90.

Kumar RA, Sridevi K, Kumar NV, Nanduri S, Rajagopal S. Anticancer and immunostimulatory compounds from Andrographis paniculata. J Ethnopharmacol 2004;92(2-3):291-5.

Awan AJ, Ahmed CB, Uzair M, Aslam MS, Farooq U, Ishfaq K. Family Acanthaceae and Genus Aphelandra: Ethnopharmacological and phytochemical review. Int J Pharm Pharm Sci 2014;6(10):44-50.

Negi AS, Kumar JK, Luqman S, Shanker K, Gupta MM, Khanuja SP. Recent advances in plant hepatoprotectives: A chemical and biological profile of some important leads. Med Res Rev 2008;28(5):746-72.

Roxas M, Jurenka J. Colds and influenza: A review of diagnosis and conventional, botanical, and nutritional considerations. Altern Med Rev 2007;12(1):25-48.

Kligler B, Ulbricht C, Basch E, Kirkwood CD, Abrams TR, Miranda M, et al. Andrographis paniculata for the treatment of upper respiratory infection: A systematic review by the natural standard research collaboration. Explore 2006;2:25-9.

Huang CJ, Wu MC. Differential effects of foods traditionally regarded as â€˜heatingâ€™ and â€˜coolingâ€™ on prostaglandin E2 production by a macrophage cell line. J Biomed Sci 2002;9:596-606.

Chao WW, Kuo YH, Li WC, Lin BF. The production of nitric oxide and prostaglandin E2 in peritoneal macrophages is inhibited by Andrographis paniculata, Angelica sinensis and Morus alba ethyl acetate fractions. J Ethnopharmacol 2009;122(1):68-75.

Mandal SC, Dhara AK, Maiti BC. Studies on psychopharmacological activity of Andrographis paniculata extract. Phytother Res 2001;15(3):253-6.

Salna KP, Sreejith K, Uthiralingam M, Prince MA, Milton J, Fleming AT. A comparative study of phytochemicals investigation of Andrographis paniculata and Murraya koenigii. Int J Pharm Pharm Sci 2011;3(3):291-2.

Rao YK, Vimalamma G, Rao CV, Tzeng Y. Flavonoids and andrographolides from Andrographis paniculata. Phytochemestry 2004;65:2317-21.

Xu C, Chou GX, Wang ZT. A new diterpene from the leaves of Andrographis paniculata Nees. Fitoterapia 2010;81(6):610-3.

Abidin SZ, Liew WL, Ismail S, Chan KL, Mahmud R. Inhibitory effects of active constituents and extracts of Andrographis paniculata on Ugt1a1, Ugt1a4, and Ugt2b7 enzyme activities. Int J Pharm Pharm Sci 2014;6(6):58-66.

Kishore PH, Reddy MV, Reddy MK, Gunasekar D, Caux C, Bodo B. Flavonoids from Andrographis lineata. Phytochemistry 2003;63(4):457-61.

Bhaskar Reddy MV, Kishore PH, Rao CV, Gunasekar D, Caux C, Bodo B. New 2â€™-oxygenated flavonoids from Andrographis affinis. J Nat Prod 2003;66(2):295-7.

Kuroyanagi M, Sato M, Ueno A, Nishi K. Flavonoids from Andrographis paniculata. Chem Pharm Bull 1987;35:4429-35.

Sukardiman S, Suharjono, Balqianur T. The role of ethyl acetate fraction of Andrographis paniculata and doxorubin combination toward the increase of apoptosis and decrease of VEGF protein expression of mice fibrosarcoma cells. Int J Pharm Pharm Sci 2015;7(4):347-50.

Cheung HY, Cheung CS, Kong CK. Determination of bioactive diterpenoids from Andrographis paniculata by micellar electrokinetic chromatography. J Chromatogr A 2001;930(1-2):171-6.

Pholphana N, Rangkadilok N, Thongnest S, Ruchirawat S, Ruchirawat M, Satayavivad J. Determination and variation of three active diterpenoids in Andrographis paniculata (Burm.f.) Nees. Phytochem Anal 2004;15(6):365-71.

Burgos RA, Caballero EE, SÃ¡nchez NS, Schroeder RA, Wikman GK, Hancke JL. Testicular toxicity assessment of Andrographis paniculata dried extract in rats. J Ethnopharmacol 1997;58(3):219-24.

Ragasa CY, de los Santos A, Rideout JA. An antimicrobial and cytotoxic labdane diterpene from Andrographis paniculata. ACGC Chem Res Commun 2008;22:44-8.

Tan MC, Oyong GG, Shen CC, Ragasa CY. Chemical constituents of Andrographis paniculata (Burm.f.) Nees. Int J Pharmacogn Phytochem Res 2016;8:1398-402.

Tan MC, Oyong GG, Shen CC, Ragasa CY. Secondary metabolites from Andrographis paniculata (Burm.f.) Nees. Der Pharm Lett 2016;8:157-60.

Tan MC, Oyong GG, Shen CC, Ragasa CY. Chemical composition of Andrographis paniculata (Burm.f.) Nees. Res J Pharm Biol Chem Sci 2016;7:2405-8.

Freshney RI. Culture of Animal Cells: A Manual of Basic Techniques. New York, USA: Wiley-Liss Inc.; 2000.

Riss TL, Moravec RA. Use of multiple assay endpoints to investigate the effects of incubation time, dose of toxin, and plating density in cell-based cytotoxicity assays. Assay Drug Dev Technol 2004;2:51-62.

Geran RI, Greenberg NH, McDonald MM, Schumacher AM, Abbott BJ. Protocols for screening chemical agents and natural products against animal tumour and other biological systems. Cancer Chemother Rep 1972;3:17-9.

Jacinto SD, Chun EA, Montuno AS, Espineli DL, Ragasa CY. Cytotoxic cardenolide and sterols from Calatropis gigantea. Nat Prod Commun 2011;6:803-6.

Makizumi R, Yang WL, Owen RP, Sharma RR, Ravikumar TS. Alteration of drug sensitivity in human colon cancer cells after exposure to heat: Implications for liver metastatis therapy using RFA and chemotherapy. Int J Clin Exp Med 2008;1:117-29.

Ravizza R, Gariboldi MB, Passarelli L, Monti E. Role of the p53/p21 system in the response of human colon carcinoma cells to Doxorubicin. BMC Cancer 2004;4:92.

Chao HP, Kuo CD, Chiu JH, Fu SL. Andrographolide exhibits anti-invasive activity against colon cancer cells via inhibition of MMP2 activity. Planta Med 2010;16:1827-33.

Sukardiman H, Widyawaruyanti A, Sismindari, Zaini NC. Apoptosis inducing effect of andrographolide on TD-47 human breast cancer cell line. Afr J Tradit Complement Altern Med 2007;4(3):345-51.

Li J, Zhang C, Jiang H, Cheng J. Andrographolide inhibits hypoxia-inducible factor-1 through phosphatidylinositol 3-kinase/AKT pathway and suppresses breast cancer growth. Oncotargets Ther 2015;8:427-35.

Banerjee M, Chattopadhyay S, Choudhuri T, Bera R, Kumar S, Chakraborty B, et al. Cytotoxicity and cell cycle arrest induced by andrographolide lead to programmed cell death of MDA-MB-231 breast cancer cell line. J Biomed Sci 2016;23:40.

Chao CY, Li CK, Hsu YT, Lu CY, Liu KL, Li CC, et al. Induction of heme oxygenase-1 and inhibition of TPA-induced matrix metalloproteinase-9 expression by andrographolide in MCF-7 human breast cancer cells. Carcinogenesis 2013;34:1843-51.

Lin HH, Shi MD, Tseng HC, Chen JH. Andrographolide sensitizes the cytotoxicity of human colorectal carcinoma cells toward cisplatin via enhancing apoptosis pathways in vitro and in vivo. Toxicol Sci 2014;139:108-20.

Luo X, Luo W, Lin C, Zhang L, Li Y. Andrographolide inhibits proliferation of human lung cancer cells and the related mechanisms. Int J Clin Exp Med 2014;7(11):4220-5.

Lee YC, Lin HH, Hsu CH, Wang CJ, Chiang TA, Chen JH. Inhibitory effects of andrographolide on migration and invasion in human non-small cell lung cancer A549 cells via down-regulation of PI3K/Akt signaling pathway. Eur J Pharmacol 2010;632(1-3):23-32.

Lim JC, Chan TK, Ng DS, Sagineedu SR, Stanslas J, Wong WS. Andrographolide and its analogues: Versatile bioactive molecules for combating inflammation and cancer. Clin Exp Pharmacol Physiol 2012;39(3):300-10.

Roy DN, Mandal S, Sen G, Mukhopadhyay S, Biswas T. 14-Deoxyandrographolide desensitizes hepatocytes to tumour necrosis factor-alpha-induced apoptosis through calcium-dependent tumour necrosis factor receptor superfamily member 1A release via the NO/cGMP pathway. Br J Pharmacol 2010;160:1823-43.

RodrÃguez-FernÃ¡ndez E, Manzano JL, Alonso A, Almendral MJ, PÃ©rez-AndrÃ©s M, Orfao A, et al. Fluorescent cisplatin analogues and cytotoxic activity. Curr Med Chem 2009;16(32):4314-27.

Pfisterer PH, Rollinger JM, Schyschka L, Rudy A, Vollmar AM, Stuppner H. Neoandrographolide from Andrographis paniculata as a potential natural chemosensitizer. Planta Med 2010;76(15):1698-700.

Mulukuri NV, Mondal NB, Prasad MR, Renuka S, Ramakrishna K. Isolation of diterpenoid lactones from the leaves of Andrographis paniculata and its anticancer activity. Int J Pharmacogn Phytochem Res 2011;3:39-42.

Nanduri S, Nyavanandi VK, Thunuguntla SS, Kasu S, Pallerla MK, Ram PS, et al. Synthesis and structure-activity relationships of andrographolide analogues as novel cytotoxic agents. Bioorg Med Chem Lett 2004;14(18):4711-7.