Journal of Ethnopharmacology 131 (2010) 363–367 Contents lists available at ScienceDirect Journal of Ethnopharmacology journal homepage: www.elsevier.com/locate/jethpharm Constituents isolated from Cordyceps militaris suppress enhanced inflammatory mediator’s production and human cancer cell proliferation Yerra Koteswara Rao a , Shih-Hua Fang b , Wen-Shi Wu c , Yew-Min Tzeng a,∗ a Institute of Biochemical Sciences and Technology, Chaoyang University of Technology, 168 Gifeng East Road, Taichung County 413, Wufeng, Taiwan, ROC b Institute of Athletes, National Taiwan Sport University, Taichung, Taiwan, ROC c Department of Horticulture and Biotechnology, Chinese Culture University, Taipei, Taiwan, ROC article info Article history: Received 16 April 2010 Received in revised form 16 June 2010 Accepted 6 July 2010 Available online 13 July 2010 Keywords: Cordyceps militaris Phytoconstituents Inflammatory mediators Human cancer cells proliferation abstract Aim of the study: The purpose of this study is to isolate the pure compounds from the extracts of Cordy- ceps militaris obtained through solid-state cultivation process, and evaluate their anti-inflammatory and anticancer properties. Materials and methods: Silica gel column chromatographic purification of Cordyceps militaris extracts resulted in the isolation of 10 pure compounds (1–10). The compounds 1–10 were examined for their growth inhibitory properties against nitric oxide (NO), tumor necrosis factor (TNF)- and interleukin (IL)- 12 enhanced production from LPS/IFN--stimulated macrophages. Additionally, the anti-proliferation effects of 1–10 on human cancer cell lines, colon (colon 205), prostate (PC-3), and hepatoma (HepG2) cells were also analyzed. Results: Compound 8 displayed potent growth inhibition on NO, TNF- and IL-12 production with an IC 50 value of 7.5, 6.3, and 7.6 g/ml, respectively. A similar inhibitory trend on these inflammatory mediators was observed for 3, 7, 9 and 10 with an IC 50 values ranging from 10.8 to 17.2 g/ml. On the other hand, compounds 3 and 8 were potent anti-proliferative agents with an IC 50 value of 35.6 and 32.6 g/ml toward PC-3 and colon 205 cell lines, respectively. The compounds 1 and 2 showed potent anti-proliferation in PC-3 and colon 205 cells, while only 3 displayed such effect in HepG2 cells. Conclusion: The present study provides scientific supporting information for the ethnopharmacological use of Cordyceps militaris as an anti-inflammatory and anticancer agent. © 2010 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Various medicinal fungi have been traditionally used in many countries for health maintenance as well as prevention and treat- ment of various diseases. Dong-Chong-Xia-Cha in Chinese, which translates as “winter worm and summer grass”, is an entomoge- nous fungus that colonized the larvae or pupae of insect. It includes different genera, such as Cordyceps, Paecilomyces, Torrubiella, and Podonectria. Cordyceps militaris is one of the medicinal mushroom infecting lepidopteron insects and parasites and kills moth pupae. It is an ascomycete that invades the pupae in the ground and that remains a worm during the winter and transforms into a mushroom in the summer. In traditional Chinese medicine the hot water and alcoholic extracts of Cordyceps militaris have been used as a tonic for longevity, endurance and vitality for thousands of years (Ying et al., 1987). A wide range of pharmacological activities have been reported on the extracts of Cordyceps militaris include ∗ Corresponding author. Tel.: +886 4 23323000x4471; fax: +886 4 23395870. E-mail address: ymtzeng@cyut.edu.tw (Y.-M. Tzeng). immunomodulatory, anti-tumor, and anti-inflammatory (Park et al., 2005; Lee et al., 2006; Kim et al., 2008; Han et al., 2010). Human clinical trials on Cordyceps militaris demonstrated that it has efficacy for relieving the symptoms of chronic bronchitis (Zhu et al., 2006). Furthermore, previous reports also indicate that Cordyceps militaris has beneficial therapeutic effects for the treatment of influenza A viral infections (Ohta et al., 2007). Because of the over exploitation, there is a scarcity for natu- ral Cordyceps. However, in the commercial market the price of this medicinal fungus has been increased by ∼20-fold during the last 10 years. Therefore, artificial methods include solid-state cultiva- tion process are developed for the production of Cordyceps militaris fruiting bodies. Previously, it was reported that the pharmacolog- ical efficacy of both cultivated and natural Cordyceps militaris are comparable and cultivated products are commonly sold as drug materials and healthy food materials in China and South East Asia (Ying et al., 1987; Zhu et al., 2006). Further, in human clinical trials both naturally grown and cultivated mycelia of Cordyceps are effective for the treatment of chronic bronchitis, asthma and other respiratory diseases (Han, 1995). The anti-oxidant activity of cultured Cordyceps mycelia is also comparable to that of the 0378-8741/$ – see front matter © 2010 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2010.07.020