ANTIMYCOBACTERIAL POLYACETYLENES FROM LEVISTICUM OFFICINALE 681 Copyright © 2008 John Wiley & Sons, Ltd. Phytother. Res. 22, 681–684 (2008) DOI: 10.1002/ptr Copyright © 2008 John Wiley & Sons, Ltd. PHYTOTHERAPY RESEARCH Phytother. Res. 22, 681–684 (2008) Published online 18 March 2008 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ptr.2408 Antimycobacterial Polyacetylenes from Levisticum officinale Andreas Schinkovitz 1,2 , Michael Stavri 3 , Simon Gibbons 3 and Franz Bucar 1 * 1 Institute of Pharmaceutical Sciences, Department of Pharmacognosy, Karl-Franzens-University, Graz, Austria 2 Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy University of Illinois at Chicago, Chicago, IL, USA 3 Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy, University of London, UK No conflicts of interest concerning financial matters or personal relationships exist between the authors and those who might bias this work. The present work is in part included the PhD thesis of A. Schinkovitz (University of Graz) but has not been published elsewhere previously. The dichloromethane extract of the roots of Levisticum officinale L. (Apiaceae) exhibited significant anti- mycobacterial activity against Mycobacterium fortuitum and Mycobacterium aurum in a microtiter plate dilution assay and was further analysed following a bioassay-guided fractionation strategy. 3(R)-Falcarinol (3(R)-(-)-1,9- heptadecadien-4,6-diin-3-ol] and 3(R)-8(S)-falcarindiol [3(R)-8(S)-(+)-1,9-heptadecadien-4,6-diin-3,8-diol] could be identified as the active components in this extract. The minimal inhibitory concentration (MIC) of 3(R)-falcarinol against M. fortuitum and M. aurum was 16.4 μM while that of 3(R)-8(S)-falcarindiol was 30.7 μM against M. fortuitum and 61.4 μM against M. aurum, respectively. Previously, 3(R),8(R)-dehydrofalcarindiol was isolated from Artemisia monosperma and surprisingly this polyacetylene exhibited no antimycobacterial activity at 128 μg/mL. This indicates that the terminal methyl group is vital for retention of antimycobacterial activity. Reference antibiotics ethambutol and isoniazid exhibited an activity of 115.5 μM and 14.6 μM against M. fortuitum, and 3.4 μM and 29.2 μM against M. aurum, respectively. Copyright © 2008 John Wiley & Sons, Ltd. Keywords: Levisticum; Mycobacterium; polyacetylene; falcarinol; falcarindiol. Received 19 September 2007 Accepted 29 September 2007 INTRODUCTION Causing 1.6 million deaths annually, tuberculosis and mycobacterial related diseases still represent a serious health problem. One third of the world’s population is latently infected, of which 5 –10% will develop a clinical disease. Mycobacterium tuberculosis represents the most prominent pathogen among mycobacteria but additionally rapidly growing mycobacteria are increas- ingly being recognized as human pathogens. In addition to M. tuberculosis, these rapidly growing species are capable of causing serious lung infections, particularly in immune comprised people such as HIV-positive individuals. Furthermore, they can infect skin, spleen, bones and kidney (Lakely et al., 2001). Plant extracts and their ingredients are known to exhibit significant antimycobacterial effects and there- fore might be a valuable source of lead structures for the development of new antibiotics (Gautam et al., 2007; Gibbons, 2005). In the course of our studies on antimycobacterial plant constituents (Schinkovitz et al., 2003; Stavri et al., 2005), the roots of Levisticum officinale L. (Apiaceae), which are traditionally used to treat lung disorders, were analysed. MATERIAL AND METHODS General experimental procedures. Optical rotation was measured on a Perkin Elmer 241 MC polarimeter in dichloromethane at 589 nm. 1 H NMR and 13 C NMR (125 MHz, CDCl 3 ) data were recorded on a Bruker AVANCE 500. APCI mass spectra were recorded on a Finnigan Navigator instrument. GC-MS analysis was performed on an HP 5890 Series II Plus gas chromatograph with an HP-5MS column (0.25 mm i.d. × 30 m, film thick- ness: 0.5 μm) in combination with an HP 5989B mass spectrometer; carrier gas, helium 5.6 at a flow rate of 1.0 mL/min; injector temperature, 280 °C; detector temperature, 280 °C; heating program, 100 °C; heating rate, 8 °C/min up to 280 °C, constantly held for 20 min. Column chromatography was performed on silica gel 60 (Merck, Darmstadt) and on Sephadex LH-20 (Pharmacia Biotech, Uppsala). TLC analyses were car- ried out on silica gel 60 F 254 pre-coated plates (Merck, Darmstadt). Spots were visualized by spraying with vanillin–sulphuric acid reagent followed by heating for 5 min at 100 °C. Preparation and determination of Mosher’s esters was conducted as previously described (Lechner et al., 2004). Plant material. Plant material was obtained as a com- mercial sample (Kottas, Vienna) and identity was verified by microscopy and TLC experiments. A voucher specimen is kept at the herbarium of the Institute of Pharmaceutical Sciences, Department of Pharmacognosy, University of Graz. * Correspondence to: Ao. Univ.-Prof. Dr Franz Bucar, Institute of Phar- maceutical Sciences; Department of Pharmacognosy, Karl-Franzens- University Graz; Universitaetsplatz 4/1A-8010 Graz, Austria. E-mail: franz.bucar@uni-graz.at