Cytotoxic and Anti-oxidant Activities of Lanostane-Type Triterpenes Isolated from Poria cocos

Abstract

NotesPoria cocos (P. cocos), which is also known as Fu Ling, is often used in Chinese traditional medicine for its diuretic and sedative effects.1) Compounds with anti-cancer properties have been isolated from both the aqueous and alcoholic extracts of P. cocos in various studies.2-7) Polysaccharides from aqueous extracts have also been shown to exhibit various anti-tumor and cytotoxic effects against certain cancer cell lines.8-10) Likewise, lanostane-type triterpenes from alcoholic extracts were shown to possess certain anti-cancer properties, including inhibition of DNA polymerases, 4,11) inhibition of tumor cell growth induced by TPA (12-O-tetradecanoylphorbol-13-acetate), 2,12,13) and inhibition of DNA topoisomerase II.4) Cytotoxicity of lanostane-type triterpenes against certain human cancer cell lines have also been reported. 2,3,14) Recently, we have shown that pachymic acid (1), a lanostane-type triterpene found in P. cocos alcoholic extract, is effective in preventing the growth of both androgenresponsive and -insensitive prostate cancer cells by inducing apoptosis in a dose-and time-dependent fashion.5) This finding, together with the observation that various lanostane-type triterpenes have been isolated from this plant, have motivated us to conduct further research on P. cocos to identify other pachymic acid-like compounds with equal or greater cytotoxicity. We report here the isolation and characterization of one novel and eight known lanostane-type triterpenes from the sclerotia of P. cocos, and the cytotoxic and antioxidant activity of these triterpenes. Results and DiscussionTo isolate compounds from P. cocos, dry sclerotia of P. cocos were crushed and extracted using 95% ethanol at 60°C under reflux. Subsequently, the alcoholic extracts were dried and fractionated using flash column chromatography. In all, four fractions, PC A-D, were obtained and evaluated for their cytotoxic activity against DU145 and A549 cells. Of these fractions, PCB gave the best response, which, at 37 mg/ml, caused cell viability of DU145 and A549 to decrease to 67.7% and 44%, respectively, after 72 h treatment (data not shown to conserve space). This was followed by PCC, which had slightly lower cytotoxicity against DU145 growth (84.7% vs. 67.7% growth inhibition at 37 mg/ml over a 72 h treatment period) relative to PCB but similar inhibiting activity as PCB on A549 cells (data not shown). PCB and PCC were then subjected to repeated column chromatography to isolate pure compounds. In all, nine lanostane-type triterpenes, including pachymic acid (1), 15) dehydropachymic acid (2), 15) 3-acetyloxy-16a-hydroxytrametenolic acid (3), 16) polyporenic acid C (4), 4) 3-epi-dehydropachymic acid (5), 16) 3-epi-dehydrotumulosic acid (6), 3,17) tumulosic acid (7), 29-hydroxypolyporenic acid C (8), and dehydrotumulosic acid (9) 17) were isolated and identified. Identification of known compounds was conducted by comparison of their physical and spectroscopic data ( 1 H-, 13 C-NMR and MS) with the corresponding compounds reported in the...

Advanced ultra-performance liquid chromatography–photodiode array–quadrupole time-of-flight mass spectrometric methods for simultaneous screening and quantification of triterpenoids in Poria cocos

“…The observed trends of the 5 identified markers may help understanding the clustering of the samples from different origins. In addition, being reported to possess interesting bioactivities (Gapter et al, 2005;Kang et al, 2006;Sato et al, 2002;Zhou et al, 2008), these 5 markers can be used as good reference compounds for quality control of P. cocos.…”

“…The main chemical constituents of P. cocos are polysaccharides, triterpenoids, egosterol and proteins (Tai, Akahori, & Shingu, 1992;Tai, Shingu, Kikuchi, Tezuka, & Akahori, 1995a;Tai, Shingu, Kikuchi, Tezuka, & Akahori, 1995b;Wang & Wan, 1998;Yang & Bao, 2005), in which triterpenoids are found to possess interesting pharmacological actions such as cytotoxic and anti-oxidant (Zhou et al, 2008), anti-cancer (Gapter, Wang, Glinski, & Ng, 2005), anti-inflammatory (Yasukawa et al, 1998), inhibition of free radical-induced lysis of red blood cells (Sekiya, Goto, Shimada, & Terasawa, 1998) and inhibition of calf DNA polymerase a and rat DNA polymerase b (Akihisa et al, 2004;Kaminaga et al, 1996). Up to now, more than 80 triterpenoid compounds were isolated and identified from P. cocos with the above-mentioned bioactivities.…”

Triterpenes from Poria cocos suppress growth and invasiveness of pancreatic cancer cells through the downregulation of MMP-7

“…As previously demonstrated P. cocos triterpenes: PA, DPA and PPAC inhibited growth of human breast cancer, lung cancer and prostate cancer cells (11)(12)(13)22). To evaluate whether these triterpenes also affect growth of different human pancreatic cancer cell lines, Panc-1, MiaPaca-2, AsPc-1 and BxPc-3 cells were treated with PTE (0-80 µg/ml) for 24 and 48 h and the proliferation determined as described in Materials and methods.…”

“…Part of the PEB (PTE) was subjected to high-performance preparative liquid chromatography (Daojing, Japan, model: SPD-20A), from which three pure compounds, PPAC, DPA and PA (Fig. 1) 13 C -NMR and MS) with the corresponding compounds reported in the literatures. PTE, PA, DPA and PPAC were dissolved in DMSO at a concentration of 50 mg/ml and 50 mM, respectively then stored at -20˚C.…”

“…Poria cocos (also known as Wolfiporia extensa) is a medicinal mushroom in the Polyporaceae family that grows in pine trees and its sclerotium is widely used in traditional Asian medicine for its sedative, diuretic, digestive and tonic effects (6)(7)(8). Although the anticancer activity of polysaccharides extracted from P. cocos is associated with the stimulation of immune response and these polysaccharides significantly enhance immunopotentiation (9), triterpenes isolated from P. cocos have a direct inhibitory effect on cancer cells through a variety of mechanisms including inhibition of cell proliferation, induction of apoptosis and suppression of invasive behavior (10)(11)(12)(13)(14)(15)(16). However the effect of triterpenes from P. cocos against pancreatic cancer remains to be evaluated and the mechanism determined.…”