METHYL 10-EPI-PHEOPHORBIDE A FROM MCF-7 CELLS ACTIVE LAYER OF THE INDONESIAN FICUS DELTOIDEA JACK LEAVES
Keywords:Ficus deltoidea, NMR, HRMS, Cytotoxic, Zebrafish embryos, Alkaloid, Chlorin, Conformational analysis
Objective: To isolate and elucidate a cytotoxic principle against breast tumor MCF-7 cells of the Indonesian terrestrial plant Ficus deltoidea Jack leaves.
Methods: F. deltoidea leaves collected at National Park of mount Gede-Pangrango, Indonesia have been subjected to chemical and biological work. F. deltoidea leaves were extracted with 96% aqueous ethanol (EtOH) and was then partitioned into three layers n-hexane, dichloromethane (CH2Cl2), and n-butanol (n-BuOH). All layers were checked for their activity against breast tumor MCF-7 cells using MTT assay method. A portion of the most active layer was purified using open column chromatography to give fraction that has toxicity against zebra fish embryos. Based on the assay-guided isolation, compound 1 was isolated. The chemical structure of 1 was elucidated using nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS) data as well as comparing data with literature.
Results: The CH2Cl2 layer of F. deltoidea leaves was found to inhibit breast tumor MCF-7 cells with IC50 10 Âµg/ml which was the most toxic among the layers. A portion of the most active layer was purified using open column chromatography to give 7 fractions. The fraction 5 showed toxicity against zebrafish embryos (LC50 35 Âµg/ml, 48 hpf). This fraction was purified using high performance liquid chromatography (HPLC) octadecylsilyl (ODS) column with gradient elution 70% aqueous acetonitrile (MeCN) to 100% MeCN (linear gradient) for 40 min with UV detection at 254 nm (tR = 30.99 min) to give compound 1. The chemical structure of 1 was revealed as a chlorin-type compound named methyl 10-epi-pheophorbide A.
Conclusion: Methyl 10-epi-pheophorbide A was isolated for the first time from the active fraction of the Indonesian F. deltoidea leaves or tabat barito. The chemical structure including absolute stereochemistry was elucidated using NMR and HRMS data as well as by comparison with the literature values. The 13C NMR data has been added to complete the previous report.
Loutfy MHA, Karakish EAK, Khalifa SF, Mira ERA. Numerical taxonomic evaluation of leaf architecture of some species of genus Ficus L. Int J Agric Biol 2005;7:352-357.
Bunawan H, Amin NM, Bunawan SN, Baharum SN, Noor NM. Ficus deltoidea Jack: a review on its phytochemical and pharmacological importance. Evid Based Complement Alternat Med 2014:902734.
Hakiman M, Maziah M. Non-enzymatic and enzymatic antioxidant activities in aqueous extract of different Ficus deltoidea accessions. J Med Plant Res 2009;3:120-131.
Omar MH, Mullen W, Crozier A. Identification of proanthocyanidin dimers and trimmers, flavone C- glycosides, and antioxidants in Ficus deltoidea, a Malaysian herbal tea. J Agric Food Chem 2011;59:1363-1369.
Rosnah J, Khandaker MM, Boyce AN. Ficus deltoidea: review on background and recent pharmacological potential. J Agron 2015;14:310-318.
Zunoliza A, Khalid H, Zhari I, Rasadah MA, Mazura P, Fadzureena J, Rohana SE. Evaluation of extracts of leaf of three Ficus deltoidea varieties for antioxidant activities and secondary metabolites. Pharmacog Res 2009;1(4):216-223.
Zakaria ZA, Hussain MK, Mohamad AS, Abdullah FC, Sulaiman MR. Antiinflammatory activity of the aqueous extract of Ficus deltoidea. Biol Res Nurs 2012;14:90-97.
Sulaiman MR, Hussain MK, Zakaria ZK, Somchit MN, Moin S, Mohamad AS, Israf DA. Evaluation of the antinociceptive activity of Ficus deltoidea aqueous extract. Fitoterapia 2008;79:557-561.
Uyub AM, Nwachukhu IN, Azlan AA, Fariza SS. In vitro antibacterial activity and cytotoxicity of selected medicinal plant extracts from Penang island Malaysia on metronidazole-resistant helicobacter pylori and some pathogenic bacteria. Ethnobotany Research & Application 2010;8:95-106.
Samah OA, Tarwiyah N, Zaidi A, Sule AB. Antimicrobial activity of Ficus deltoidea Jack. (Mas Cotek). Pak J Pharm Sci 2012;25:675-678.
Suryati, Nurdin H, Dachriyanus, Lajis MNH. Structure elucidation of antibacterial compound from Ficus deltoidea Jack leaves. Indones J Chem 2011;11:67-70.
Oh MJ, Hamid MA, Ngadiran S, Seo YK, Sarmidi MR, Park CS. Ficus deltoidea (Mas cotek) extract exerted anti-melanogenic activity by preventing tyrosinase activity in vitro and by suppressing tyrosinase gene expression in B16 F1 melanoma cells. Arch Dermatol Res 2011; 303:161-170.
Sakdarat S, Shuyprom A, Pientong C, Ekalaksananan T, Thongchai S. Bioactive constituents from the leaves of Clinacanthus nutans Linda. Bioorg Med Chem 2009;17:1857-1860.
Wua SJ, Ng LT, Wang GH, Huang YJ, Chen JL, Sun FM. Chlorophyll a, an active anti-proliferative compound of Ludwiga octovalvis, activates the CD95 (APO-1/CD-95) system and AMPK pathway in 3T3-L1 cells. Food Chem Toxicol 2010;48:716-721.
Lip JM, Hisham DN, Zaidi JA, Musa Y, Ahmad AW, Normah A , Sharizan A. Isolation and identification of moretenol from Ficus deltoidea leaves. J Trop Agric and Fd Sc 2009; 37:195-201.
Choo CY, Sulong NY, Man F, Wong TW. Vitexin and isovitexin from the leaves of Ficus deltoidea with in-vivo Î±-glucosidase inhibition. J Etnopharmacol 2012;142:776-781.
Lin HY,Chiu HL, Lu TL, Tzeng CY, Lee TH, Lee CK, Shao YY, Chen CR, Chang CI, Kuo YH. Ficusmicrochlorin A-C, two new methoxy lactone chlorins and an anhydride chlorin from the leaves of Ficus microcarpa. Chem Pharm Bull 2011;59:113-116.
Lin YH, Chiu HL, Lan YH, Tzeng CY, Lee TH, Lee CK, Shao YY, Chen CR, Chang CI, Kuo YH. Ficuschlorins A-D, lactone chlorins from the leaves of Ficus microcarpa. Chem Biodivers 2011;8:1701-1706.
Ferruzzi MG, Blakeslee J. Digestion, absorption, and cancer preventative activity of dietary chlorophyll derivatives. Nutr Res 2007;27:1-12.
Wongsingkongman P, Brossi A, Wang HK, Bastow KF, Lee KH. Antitumor agents. Part 209: Pheophorbide-a derivatives as photo-independent cytotoxic agents. Bioorg Med Chem 2002;10:583-591.
Chan JY, Yang PM, Hon PM, An SW, Tsui SK, Wave MM. Pheophorbide a, a major antitumor component purified from Scutellaria barbata, induces apoptosis in human hepatocellular carcinoma cells. Planta Med 2006;72:28-33.
Hanif N, Irawadi TT, Affandi H, Read RW, Nomura T. Artonol B, an isoprenylated phenol from the stem-bark of Terap (Artocarpus anisophyllus Miq). Presented at National Symposium on Natural Product Chemistry VII, Depok, Indonesia, 16-17 November 1999, 386-390.
Matsuo A, OnoK, Hamasaki K, Nozaki H. Phaeophytins from a cell suspension culture of the liverwort Plagiochila ovalifolia. Phytochemistry 1996;42:427-430.
Nakatani Y, Ourisson G, Beck JP. Chemistry and biochemistry of Chinese drugs VII: cystostatic phephytins from silkworm excreta, and derived photocytotoxic pheophorbides. Chem Pharm Bull 1981;29:2261-2269.
Wolf H, Brockmann HJr, Biere H, Inhoffen HH. Darstellung der diastereomeren 10-Methoxy-(pyro)-methyl- phaophorbide a und bestimmung der relativen Konfiguration am C-Atom10. Liebigs Ann Chem 1967;704:208-225.
Ellsworth PA, Storm CB. Methyl 10-ephipheophorbide a:an unusual epimeric stability relative to chlorophyll a or aâ€™. J Org Chem 1978;43:281-283.