THE INVESTIGATION OF BIOACTIVE
Anogeissus rivularis (COMBRETACEAE)
COMPOUNDS
FROM
ROOTS
OF
Wannee Sukbangnop1,*, Vichai Reutrakul1, Patoomratana Tuchinda1, Chutima Kuhakarn1,
Natthinee Anantachoke2, Pawinee Piyachaturawat3, Surawat Jariyawat3 and Kanoknetr Suksen3
1Department
of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of
Science, Mahidol University, Bangkok 10400, Thailand
2
Department of Pharmacognosy, Faculty of Pharmacy, Mahidol University, Bangkok 10400,
Thailand
3
Department of Physiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
*e-mail: wanneesukbangnop@gmail.com, #e-mail: vichai.reu@mahidol.ac.th.
Abstract:
Seven known compounds, betulinic acid (1), β-sitosterol (2), 3,3',4-tri-O-methylflavellagic acid (3) and 3,3'-di-O-methylellagic acid (4) pterostilbene (5), termilignan B (6) and
3',5',5,7-tetrahydroxyflavanone (7) were isolated from the methanol extract of roots of
Anogeissus rivularis (Combretaceae). The structures of 1-7 were determined on the basis of
spectroscopic methods and were confirmed by comparison with the data previously reported in
the literatures. Compound 1 exhibited significant cytotoxic activity against P-388 cell line.
Keywords: Anogeissus rivularis, betulinic acid, β-sitosterol, pterostilbene, termilignan B, 3,3',4-tri-Omethylflavellagic acid and 3,3'-di-O-methylellagic acid
Introduction
The Combretaceae family comprises 20 genera and more than 600 species. Combretum
and Terminalia are the largest genera, with over 200 species each. The investigation by
Smittinard indicated the presence of 6 genera, namely Anogeissus Combretum, Getonia,
Lumnitzera, Quisqualis and Terminalia.1 Anogeissus is a genus of trees, native to South Asia and
Africa,4 the Arabian Peninsula, and Africa, belonging to family Combretaceae.2 This genus was
elevated from the section of Conocarpus and distinguished from Conocarpus by the fruits.3
Anogeissus fruits are aggregated into con-like heads with wings or ribs and a beak at the apex,
representing the persistent calyx-stalk. Conocarpus fruits have a deciduous calyx-stalk. The
genus Anogeissus comprises 19 species. In Thailand, only two species are found, namely
A. acuminata (Roxb. Ex DC.) Guill. & Perr. var. lanceolata and A. rivularis (Gagnep)
O.
Lecompte [ST].1 Anogeissus rivularis is a shrub or small tree, growing up to 15 m with limb
weakness, leaf stalk bract, stem and outer tubes sepals short with soft hairs. It is found in Laos
and Thailand. In Thailand, it is found in the district of Khong Chiam (Ubon Ratchathani
province) and Phichit province. Anogeissus rivularis is called by local name, “Takhian Narm”
(ต ะ เคี ย น น้ ำ ) in Ubon Ratchathani province and “Cro-Thien” ( ค ร่ อ เที ย น ) in Phichit province.
Therefore, we are interested in the isolation and structural elucidation of the chemical
constituents from roots of this plant. The separated fractions (F1-F9) from Anogeissus rivularis
were evaluated for their biological activities.
Methodology
The air-dried and finely powdered roots of Anogeissus rivularis were extracted with
methanol at room temperature, followed by filtration. The filtrates were combined and
evaporated to dryness under reduced pressure to give a crude methanol extract. The crude extract
was taken for biological assays, i.e. cytotoxic assay with various mammalian cancer cell lines.
The crude methanol extract was dissolved with EtOAc:MeOH (1:1) at room temperature,
followed by MeOH. The crude EtOAc:MeOH (1:1) fraction was first separated by column
chromatography to afford 9 subfractions. Fraction F4 was further separated by column
chromatography to give β-sitosterol (2) and termilignan B (6). Fraction F5 was further separated
by column chromatography to give betulinic acid (1) and pterostilbene (5). Fraction F7 was
further separated by column chromatography to give 3,3',4-tri-O-methylflavellagic acid (3) and
3,3'-di-O-methylellagic acid (4). Fraction F8 was further separated by column chromatography to
give 3',5',5,7-tetrahydroxyflavanone (7).
Results and Discussion
Seven known compounds, betulinic acid (1), β-sitosterol (2), 3,3',4-tri-O-methylflavellagic acid (3) and 3,3'-di-O-methylellagic acid (4) pterostilbene (5), termilignan B (6) and
3',5',5,7-tetrahydroxyflavanone (7) were isolated from the EtOAc:MeOH (1:1) fraction of roots
of Anogeissus rivularis. The known compounds were identified by comparison of their physical
properties and spectroscopic data with those reported in the literature.5-10
Pure compounds 1 and 2 were evaluated for cytotoxic effects against a panel of cultured
mammalian cancer cell lines and non-cancerous human embryonic kidney cell (Hek 293). The
results are as shown in Table 1. Compound 1 exhibited cytotoxic activity against P-388 cell line.
while compounds 2 did not show any activity. The cytotoxic activities of compounds 3-7 will be
further evaluated.
Table 1. Cytotoxicities of compounds 1 and 2
Compounds
1
2
P-388
3.59
-
KB
-
Cell line (ED50 µg/mL)a
HT29
MCF-7
A549
-
ASK
-
Hek 293
9.29
-
a
Results are expressed as ED50 (µg/mL): ED50 < 4 µg/mL is considered active for pure compound. *P-388 = Murine lymphocytic leukemia, KB
= Human nasopharyngeal carcinoma, HT29 = Human colorectal adenocarcinoma, MCF-7 = Human breast cancer, A549 = Human lung
carcinoma, ASK = Cell line from rat glioma, Hek 293= Non-cancerous human embryonic kidney cell line.
Conclusion
In conclusion, seven known compounds were isolated from roots of Anogeissus rivularis.
The cytotoxicities of compounds 1 and 2 are presented.
Fig1. The structures of compounds 1-7 isolated from roots of Anogeissus rivularis.
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Acknowledgement:
This research project is supported by Mahidol University, the Office of the Higher Education Commission
and Mahidol University under Higher Education Research Promotion (HERP), the National Research Universities
(NRU) and Center of Excellence for Innovation in Chemistry (PERCH-CIC).