Journal of Ethnopharmacology 121 (2009) 292–296 Contents lists available at ScienceDirect Journal of Ethnopharmacology journal homepage: www.elsevier.com/locate/jethpharm The constituents of Anisomeles indica and their anti-inflammatory activities Yerra Koteswara Rao a , Shih-Hua Fang b,c , Shih-Chuan Hsieh a , Tsung-Hsien Yeh a , Yew-Min Tzeng a,∗ a Institute of Biochemical Sciences and Technology, Chaoyang University of Technology, Wufeng, Taiwan, ROC b Department of Microbiology, School of Medicine, China Medical University, Taichung, Taiwan, ROC c Institute of Athletics, National Taiwan College of Physical Education, Taichung, Taiwan, ROC article info Article history: Received 20 August 2008 Received in revised form 8 October 2008 Accepted 30 October 2008 Available online 8 November 2008 Keywords: Anisomeles indica Labiatae Whole plants Isolation of components Anti-inflammatory activity abstract Ethnopharmacological relevance: Anisomeles indica (L.) Kuntze. (Labiatae), popularly known as ‘yu-chen- tsao’, has been traditionally used as anti-inflammatory agent. Aim of the study: Investigate the chemical constituents from the whole plants of Anisomeles indica, and evaluate their in vitro anti-inflammatory activities. Results: The combined MeOH extract was successively partitioned with CHCl 3 and n-butanol, then submit- ted to several column chromatographic, and HPLC purification procedures which led to the isolation of one cembrane-type diterpenoid (3), two benzenoids (4 and 5), five flavonoids (1, 2, 6, 7 and 14), and six phenyl propanoids (8–13). The compounds 1–14 were examined for their inhibitory effects on inflammatory mediator’s enhanced production from LPS/IFN--stimulated macrophages. Among these, ovatodiolide (3) exhibited potent inhibition on NO, TNF- and IL-12 enhanced production at a concentration of 5 M, followed by pedalitin (1), scutellarein 7-O--d-glucuronide methyl ester (6), and acteoside (12) at 40 M (P < 0.05). Furthermore, 2 M of 3, and 20 M of 1 and 6 significantly (P < 0.05) arrested the cell cycle of Con A-stimulated spleen cells at the G0/G1 stage. Conclusion: This is the first report on the presence of compounds 1 and 4–13 in this plant and of the potent anti-inflammatory activity of 1, 3, 6 and 12 in vitro. These compounds may account for the use of Anisomeles indica in folk medicine to treat inflammation. © 2008 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Anisomeles is a genus of the Labiatae family comprising approxi- mately eight species, several members of which enjoy a reputation in traditional medicine throughout the southern and tropical regions of Asia (Huang, 2003). In particular, Anisomeles indica Kuntze, the only species found in Taiwan, has been used for the treatment of diverse conditions such as inflammatory skin dis- eases, liver disease and protection, gastrointestinal disease and hypertension and immune system deficiencies (Kao, 1985; Huang, 2003). Previous reports on the extracts of Anisomeles indica revealed that a decoction from the pre-flowering stage leaves and stems has anti-histaminerigic, free radical scavenging, membrane sta- bilizing, and cyclooxygenase-I inhibitory activities. In addition, the above aqueous extract was also shown to have analgesic and anti-hyperalgesic activities (Dharmasiri et al., 2002, 2003). Fur- Abbreviations: CC, column chromatography; Con A, concanavalin A; IFN, inter- feron; IL, interleukin; LPS, lipopolysaccharide; NO, nitric oxide; TNF-, tumor necrosis factor-. ∗ Corresponding author. Tel.: +886 4 23323000x4471; fax: +886 4 23395870. E-mail address: ymtzeng@cyut.edu.tw (Y.-M. Tzeng). thermore, the ethanol extract of Anisomeles indica exhibited strong anti-Helicobacter pylori activity (Wang and Huang, 2005). We have recently reported that the methanol extract of Anisomeles indica could inhibit the enhanced production of inflammatory mediators, and tumor cell proliferation (Hsieh et al., 2008). Previous investigations on the chemical composition of this herb resulted in the isolation of various constituents, including ter- penoids, flavonoids, steroids and certain miscellaneous substances (Manchand and Blount, 1977; Ansari and Dobhal, 1982; Arisawa et al., 1986a,b; Rao et al., 1983a,b,c, 1984, 1985; Dobhal et al., 1988; Shahidul Alama et al., 2000), it still lacks the evidence to elucidate the active anti-inflammatory components in Anisomeles indica. Therefore, the purpose of the present study was to isolate and identify the potent anti-inflammatory components of Ani- someles indica, and investigate their preliminary anti-inflammatory mechanism. 2. Materials and methods 2.1. General The NMR spectra were recorded in a Varian Unity Inova- 600 VXR-300/51 spectrometer operating at 600 MHz ( 1 H) and 0378-8741/$ – see front matter © 2008 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2008.10.032