METHYL 10-EPI-PHEOPHORBIDE A FROM MCF-7 CELLS ACTIVE LAYER OF THE INDONESIAN FICUS DELTOIDEA JACK LEAVES

Anggia Murni; Novriyandi Hanif; Masaki Kita; Latifah K. Darusman

doi:10.22159/ijpps.2017v9i8.18750

Authors

Anggia Murni Tropical Biopharmaca Research Center, Bogor Agricultural University, Jalan Taman Kencana No. 3, Bogor 16128, Indonesia
Novriyandi Hanif Department of Chemistry, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Gedung Kimia Wing 1 Lantai 3, Jalan Tanjung Kampus IPB Darmaga, Bogor 16680, Indonesia
Masaki Kita Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8610, Japan
Latifah K. Darusman Department of Chemistry, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Gedung Kimia Wing 1 Lantai 3, Jalan Tanjung Kampus IPB Darmaga, Bogor 16680, Indonesia

DOI:

https://doi.org/10.22159/ijpps.2017v9i8.18750

Keywords:

Ficus deltoidea, NMR, HRMS, Cytotoxic, Zebrafish embryos, Alkaloid, Chlorin, Conformational analysis

Abstract

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 (t_R = 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.

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