Sains Malaysiana 51(5)(2022): 1487-1504

http://doi.org/10.17576/jsm-2022-5105-18

 

Bioassay-guided Isolation of Triterpene Compounds from Dillenia suffruticosa and Their Cytotoxic Activities against Cancer Cells

(Pengasingan Bioasai-Terarah oleh Sebatian Triterpena daripada Dillenia suffruticosa dan Aktiviti Sitotoksik Melawan Sel Kanser)

 

ARMANIA NURDIN1,2*, LATIFAH SAIFUL YAZAN1,3 & INTAN SAFINAR ISMAIL4,5

 

1Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

2Laboratory of UPM-MAKNA Cancer Research (CANRES), Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

3Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

4Laboratory of Natural Product, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

5Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

 

Received: 30 April 2020/Accepted: 8 October 2021

 

abstract

Previous studies showed the ability of Dillenia suffruticosa to inhibit the growth of cancer cells by means of cell cycle arrest and apoptosis, thus validating the traditional use of the plant in treating cancer. Therefore, the present study was designed to isolate and elucidate the bioactive compounds responsible for the anticancer properties of D. suffruticosa extract. Bioassay-guided fractionation of the most potent fractions of DCM and EtOAc extract of D. suffruticosa was performed via column chromatography followed by purification using preparative HPLC. The structures of the isolated compounds were elucidated by NMR spectroscopy. Biological experiment by MTT assay and a series of column chromatography resulted in the isolation of three triterpene compounds. However, only the structure of compound (3) was confirmed as 1-isopropenyl-4α, 4β, 8, 10, 14-pentamethyl-icosahydro-cyclopenta[a]chrysene-3, 17-diol. Another two compounds were labelled as (1) and (2), which currently are unidentified due to unsuccessfulness in the full analysis of the spectroscopic data that enable the assignment of all protons, carbons, and confirmation of the structure. Compound (2) isolated from DCM extract of D. suffruticosa was most cytotoxic towards the selected cancer cells compared to the other two compounds and selected chemotherapeutic drugs, tamoxifen, and cisplatin. The ability of the isolated compounds to inhibit the growth of cancer cells indicates that these compounds are the bioactive constituents in D. suffruticosa that is mainly responsible for the cytotoxic activities. For this reason, these isolated compounds could be used as a means for the standardisation of herbal product from D. suffruticosa.

 

Keywords: Active constituents; bioassay-guided fractionation; cancer cells; cytotoxicity; Dillenia suffruticosa; triterpenes

 

abstrak

Kajian terdahulu mendedahkan keupayaan Dillenia suffruticosa untuk menghalang pertumbuhan sel kanser melalui tangkapan kitaran sel dan apoptosis dan membuktikan penggunaan tumbuhan tersebut secara tradisi dalam mengubati kanser. Oleh itu, kajian ini dirancang untuk memencilkan dan menguraikan sebatian bioaktif yang berperanan terhadap sifat antikanser ekstrak D. suffruticosa. Fraksinasi berpandukan bioasai terhadap fraksi yang paling poten daripada ekstrak DCM dan EtOAc tumbuhan D. suffruticosa dilakukan menggunakan kromatografi turus diikuti dengan penulenan menggunakan HPLC preparatif. Struktur sebatian yang dipencilkan telah diterbitkan menggunakan spektroskopi NMR. Uji kajibiologi menggunakan asai MTT dan siri kromatografi turus menghasilkan tiga sebatian triterpenes. Walau bagaimanapun, hanya struktur sebatian (3) telah disahkan sebagai 1-isopropenyl-4α, 4β, 8, 10, 14-pentamethyl-icosahydro-cyclopenta [a] chrysene-3, 17-diol. Dua lagi sebatian yang dilabelkan sebagai (1) dan (2), dikelaskan sebagai belum dapat dikenal pasti kerana analisis penuh data spektroskopik yang membolehkan pengenalpastian semua proton, karbon dan pengesahan struktur, tidak dapat dilaksanakan. Sebatian (2) yang telah diasingkan daripada ekstrak DCM tumbuhan D. suffruticosa adalah yang paling sitotoksik terhadap sel kanser yang dipilih berbanding dengan dua sebatian lain dan drug kemoterapi terpilih, iaitu tamoksifen dan cisplatin. Keupayaan sebatian yang dipencilkan untuk menghalang pertumbuhan sel kanser menunjukkan bahawa sebatian ini adalah unsur-unsur bioaktif dalam D. suffruticosa yang berperanan terhadap ciri sitotoksik tumbuhan tersebut. Atas sebab ini, sebatian yang dipencilkan ini dapat digunakan untuk tujuan pempiawaian produk herba daripada D. suffruticosa.

 

Kata kunci: Dillenia suffruticosa; kesitotoksikan; konstituen aktif; pemecahan bioasai-terarah; sel kanser; triterpena

 

REFERENCES

Ahmad, F.B. & Holdsworth, D.K. 1995. Traditional medicinal plants of Sabah, Malaysia part III. The Rungus people of Kudat. International Journal of Pharmacognosy 33(3): 262-264.

Alsayari, A., Kopel, L., Ahmed, M.S., Soliman, H.S.M., Annadurai, S. & Halaweish, F.T. 2018. Isolation of anticancer constituents from Cucumis prophetarum var. prophetarum through bioassay-guided fractionation. BMC Complementary and Alternative Medicine 18(1): 274.

Armania, N., Yazan, L.S., Musa, S.N., Ismail, I.S., Foo, J.B., Chan, K.W., Noreen, H., Hisyam, A.H., Zulfahmi, S. & Ismail, M. 2013a. Dillenia suffruticosa exhibited antioxidant and cytotoxic activity through induction of apoptosis and G2/M cell cycle arrest. Journal of Ethnopharmacology 146(2): 525-535.

Armania, N., Yazan, L.S., Ismail, I.S., Foo, J.B., Tor, Y.S., Ishak, N., Ismail, N. & Ismail, M. 2013b. Dillenia suffruticosa inhibits proliferation of human breast cancer cell lines (MCF-7 and MDA-MB-231) via induction of G2/M arrest and apoptosis. Molecules 18(11): 13320-13339.

Aschele, C., Bergamo, F. & Lonardi, S. 2009. Chemotherapy for operable and advanced colorectal cancer. Cancer Treatment Reviews 35(6): 509-516.

Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R.L., Torre, L.A. & Jemal, A. 2018. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. A Cancer Journal Clinicians 68(6): 394-424.

Chen, J., Lee, F.S.C., Li, L., Yang, B. & Wang, X. 2007. Standardised extracts of chinese medicinal herbs: Case study of Danshen (Salvia miltiorrhiza Bunge). Journal of Food and Drug Analysis 15(4): 347-364.

Chen, T., Xu, T., Li, Y., Liang, C., Chen, J., Lu, Y., Wu, Z. & Wu, S. 2011. Risk of cardiac dysfunction with trastuzumab in breast cancer patients: a meta-analysis. Cancer Treatment Reviews 37(4): 312-320.

Colegate, S.M. & Molyneux, R.J. 2008. Bioactive Natural Products Detection, Isolation and Structural Determination. 2nd ed. Boca Raton: CRC Press. pp. 1-9.

de Marinis, F., Rossi, A., Di Maio, M., Ricciardi, S. & Gridelli, C. 2011. Treatment of advanced non-small-cell lung cancer: Italian Association of Thoracic Oncology (AIOT) clinical practice guidelines. Lung Cancer 73(1): 1-10.

Din, O.S., Dodwell, D., Winter, M.C., Mori, S. & Coleman, R.E. 2011. Current opinion of aromatase inhibitor-induced arthralgia in breast cancer in the UK. Clinical Oncology 23(10): 674-680.

El Saghir, N.S., Tfayli, A., Hatoum, H.A., Nachef, Z., Dinh, P. & Awada, A. 2011.  Treatment of metastatic breast cancer: State-of-the-art, subtypes and perspectives. Critical Reviews in Oncology/Hematology 80(3): 433-449.

Hanum, F. & Hamzah, N. 1999. The use of medicinal plant species by the Temuan Tribe of Ayer Hitam Forest, Selangor, Peninsular Malaysia. Pertanika Journal of Tropical Agricultural Sciences 22(2): 85-94.

Hess, S.C. & Monache, F.D. 1999. Divergionic acid, a triterpene from Vochysia divergens. Journal of The Brazillian Chemical Society 10(2): 104-106.

Lai, C.S., Mas, R.H.M.H., Nair, N.K., Mansor, S.M. & Navaratnam, V. 2010. Chemical constituents and in vitro anticancer activity of Typhonium flagelliforme (Araceae). Journal of Ethnopharmacology 127(2): 486-494.

Lee, Y.Y., Lee, J.W., Park, H.S., Song, T.J., Kim, M.K., Choi, C.H., Kim, T.J., Lee, J.H., Bae, D.S. & Kim, B.G. 2010. Sequence-dependent hematologic side effects of topotecan and cisplatin in persistent or recurrent cervical cancer. Gynecologic Oncology 119(1): 87-91.

Manan, A.A., Tamin, N.S.I., Abdullah, N.S., Abidin, A.Z. & Wahab, M. 2016. Malaysian National Cancer Registry Report 2007-2011. pp. 1-228.

Mat-Salleh, K. & Latiff, A. 2002. Tumbuhan Ubatan Malaysia. Bangi: Universiti Kebangsaan Malaysia.

Mfotie Njoya, E., Weber, C., Hernandez-Cuevas, N.A., Hon, C.C., Janin, Y., Kamini, M.F., Moundipa, P.F. & Guillén, N. 2014. Bioassay-guided fractionation of extracts from Codiaeum variegatum against Entamoeba histolytica discovers compounds that modify expression of ceramide biosynthesis related genes. PLoS Neglected Tropical Diseases 8(1): e2607.

Muliawan, S.Y. 2008. Effect of Dillenia suffruticosaextract on dengue virus type 2 replication. Universa Medicina 27(1): 1-5.

Perez, E.A. 2011. New treatment strategies in the management of breast cancer. European Journal of Cancer - Supplements 9(2): 22-29.

Pieters, L. & Vlietinck, A. 2005. Bioguided isolation of pharmacologically active plant components, still a valuable strategy for the finding of new lead compounds? Journal of Ethnopharmacology 100(1-2): 57-60.

Randall-Whitis, L.M. & Monk, B.J. 2007. Topotecan in the management of cervical cancer. Expert Opinion on Pharmacotherapy 8(2): 227-236.

Shaffer, R., Tyldesley, S., Rolles, M., Chia, S. & Mohamed, I. 2009. Acute cardiotoxicity with concurrent trastuzumab and radiotherapy including internal mammary chain nodes: A retrospective single-institution study. Radiotherapy and Oncology 90(1): 122-126.

Wiart, C., Mogana, S., Khalifah, S., Mahan, M., Ismail, S., Buckle, M., Narayana, A.K. & Sulaiman, M. 2004. Antimicrobial screening of plants used for traditional medicine in the state of Perak, Peninsular Malaysia. Fitoterapia 75(1): 68-73.

Wysowski, D.K., Honig, S.F. & Beitz, J. 2002. Uterine sarcoma is associated with tamoxifen use. New England Journal of Medicine 346(23): 1832-1833.

Yang, M., Sun, J., Lu, Z., Chen, G., Guan, S., Liu, X., Jiang, B., Ye, M. & Guo, D.A. 2009. Phytochemical analysis of traditional Chinese medicine using liquid chromatography coupled with mass spectrometry. Journal of Chromatography A 1216(11): 2045-2062.

 

*Corresponding author; email: armania@upm.edu.my

 

 

 

previous