A new phenanthrene derivative 3,7-dihydroxy-2,4,6-trimethoxyphenanthrene was isolated from the all plant of
, and its structure was elucidated by extensive spectral studies and chemical transformation. The compound displayed cytotoxicity against the growth of human leukemia cell lines K562 and HL-60, human lung adenocarcinoma A549, human hepatoma BEL-7402 and human stomach cancer cell lines SGC-7901 with IC
values of 14.23, 10.02, 3.42, 15.36 and 1.13 mg/ml respectively.
MS were determined on an API Qstar Pulsa LC/TOF mass spectrometer. NMR spectra were measured on a Bruker DRX-500 spectrometer with TMS as internal standard. Silica gel (200-300 mesh) was used for column chromatography and silica gel GF
for TLC (Qingdao Marine Chemical Co., China). Solvents were of industrial purity and distilled prior to use.
The whole plant of
were collected from Simao County of Yunnan Province, China in February, 2004 and identified by one of the authors, Dr. Hong Wang, School of Life Science, Yunnan University, where a voucher specimen (No.0402017) was deposited.
Extraction and isolation.
The air-dried powdered whole plant of
(20 Kg) were extracted with 95% EtOH (20 liters × 4) at room temperature. The EtOH extract was concentrated in vacuo to yield a dark brown residue (1 Kg). H
O (2.5 L) was added to the residue, and the resultant solution was extracted with petroleum ether, EtOAc and n-BuOH successively (1.5 liters ×4). The EtOAc extract (350 g) was applied to a silica gel column, eluting with petroleum ether containing increasing amounts of acetone to obtain 6 fractions. Fr. 3 (77 g) was separated to two subfractions by silica gel column chromatography (petroleum ether-acetone 4:1, 7:3). The second subfraction (47 g) was subjected to repeated column chromatography, first on silica gel (CHCl
-acetone 80:1) and then on Sephadex LH-20 (MeOH-H
O 9:1) to obtain
H-NMR (500 MHz, CD
C-NMR (125 MHz, CD
(70 eV, ret. Int., %): 300 [M
] (100), 285 (63), 253 (17), 242 (22), 214 (33), 185 (11), 150 (16); HRESIMS
: 323.0889 [M+Na]
, requires 323.0895.
Methylation of 1:
(10 mg) was methylated with CH
CO. Preparative TLC of the crude product (silica gel, petroleum ether-EtOAc 4:1) gave 2,3,4,6,7-pentamethoxyphenanthrene (
, 6 mg) as a colorless amorphous powder. EI-MS
(70 eV, ret. Int., %): 328 [M
): δ9.06 (1H, s, H-5), 7.61 (1H, d,
=9.0 Hz, H-9), 7.57 (1H,
=9.0 Hz, H-10), 7.34 (1H, s, H-1 or H-8), 7.27 (1H, s, H-8 or H-1), 4.04, 4.01, 3.99, 3.96, 3.96 (15H, each s, OMe ×5), identical to lit. values.
This investigation was supported by a grant (No. 2005DFA30670) for international collaborative research by Ministry of Science and Technology, China, the Excellent Young Teachers Program (No. 2003 192) of MOE, China and grants (No. 2003C0033M, 2000C0001P) for scientific research from Yunnan Province, China.
Delectis Florae Reipublicae Popularis Sinicae Agendae, Academiae Sinicae Edita
Flora Reipublicae Popularis Sinicae
Wu Z. Y.
Shanghai Press of Science and Technology
Liu D. L
Pang F. G.
Wang N. L.
Yao X. S.
Lin J. G.
Zhang W. G
Niu Z. Y.
Liu D. L.
Wang N. L.
Yao X. S.
Chin. Chem. Lett.
Alley M. C.
Scudiero D. A.
Hursey M. L.
Czerwinski M. J.
Fine D. L.