Pancreatic lipase inhibitory alkaloids of Murraya koenigii leaves

Volume 4. Issue 8. Pages 1023-1170. 2009 ISSN 1934-578X (printed); ISSN 1555-9475 (online)

www.naturalproduct.us

INFORMATION FOR AUTHORS Full details of how to submit a manuscript for publication in Natural Product Communications are given in Information for Authors on our Web site http://www.naturalproduct.us. Authors may reproduce/republish portions of their published contribution without seeking permission from NPC, provided that any such republication is accompanied by an acknowledgment (original citation)-Reproduced by permission of Natural Product Communications. Any unauthorized reproduction, transmission or storage may result in either civil or criminal liability. The publication of each of the articles contained herein is protected by copyright. Except as allowed under national “fair use” laws, copying is not permitted by any means or for any purpose, such as for distribution to any third party (whether by sale, loan, gift, or otherwise); as agent (express or implied) of any third party; for purposes of advertising or promotion; or to create collective or derivative works. Such permission requests, or other inquiries, should be addressed to the Natural Product Inc. (NPI). A photocopy license is available from the NPI for institutional subscribers that need to make multiple copies of single articles for internal study or research purposes. To Subscribe: Natural Product Communications is a journal published monthly. 2009 subscription price: US$1,695 (Print, ISSN# 1934-578X); US$1,395 (Web edition, ISSN# 1555-9475); US$2,095 (Print + single site online). Orders should be addressed to Subscription Department, Natural Product Communications, Natural Product Inc., 7963 Anderson Park Lane, Westerville, Ohio 43081, USA. Subscriptions are renewed on an annual basis. Claims for nonreceipt of issues will be honored if made within three months of publication of the issue. All issues are dispatched by airmail throughout the world, excluding the USA and Canada.

NPC Natural Product Communications

EDITOR-IN-CHIEF

DR. PAWAN K AGRAWAL Natural Product Inc. 7963, Anderson Park Lane, Westerville, Ohio 43081, USA [email protected] EDITORS

PROFESSOR ALESSANDRA BRACA Dipartimento di Chimica Bioorganicae Biofarmacia, Universita di Pisa, via Bonanno 33, 56126 Pisa, Italy [email protected]

PROFESSOR DEAN GUO State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China [email protected]

PROFESSOR J. ALBERTO MARCO Departamento de Quimica Organica, Universidade de Valencia, E-46100 Burjassot, Valencia, Spain [email protected]

PROFESSOR YOSHIHIRO MIMAKI School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Horinouchi 1432-1, Hachioji, Tokyo 192-0392, Japan [email protected]

PROFESSOR STEPHEN G. PYNE Department of Chemistry University of Wollongong Wollongong, New South Wales, 2522, Australia [email protected]

PROFESSOR MANFRED G. REINECKE Department of Chemistry, Texas Christian University, Forts Worth, TX 76129, USA [email protected]

PROFESSOR WILLIAM N. SETZER Department of Chemistry The University of Alabama in Huntsville Huntsville, AL 35809, USA [email protected]

PROFESSOR YASUHIRO TEZUKA Institute of Natural Medicine Institute of Natural Medicine, University of Toyama, 2630-Sugitani, Toyama 930-0194, Japan [email protected]

PROFESSOR DAVID E. THURSTON Department of Pharmaceutical and Biological Chemistry, The School of Pharmacy, University of London, 29-39 Brunswick Square, London WC1N 1AX, UK [email protected]

ADVISORY BOARD Prof. Berhanu M. Abegaz Gaborone, Botswana

Prof. Viqar Uddin Ahmad Karachi, Pakistan

Prof. Øyvind M. Andersen Bergen, Norway

Prof. Giovanni Appendino Novara, Italy

Prof. Yoshinori Asakawa Tokushima, Japan

Prof. Lee Banting Portsmouth, U.K.

Prof. Anna R. Bilia Florence, Italy

Prof. Maurizio Bruno Palermo, Italy

Prof. Josep Coll Barcelona, Spain

Prof. Geoffrey Cordell Chicago, IL, USA

Prof. Samuel Danishefsky New York, NY, USA

Prof. Duvvuru Gunasekar Tirupati, India

Prof. A.A. Leslie Gunatilaka Tucson, AZ, USA

Prof. Stephen Hanessian Montreal, Canada

Prof. Kurt Hostettmann Lausanne, Switzerland

Prof. Martin A. Iglesias Arteaga Mexico, D. F, Mexico

Prof. Jerzy Jaroszewski Copenhagen, Denmark

Prof. Leopold Jirovetz Vienna, Austria

Prof. Teodoro Kaufman Rosario, Argentina

Prof. Norbert De Kimpe Gent, Belgium

Prof. Hartmut Laatsch Gottingen, Germany

Prof. Marie Lacaille-Dubois Dijon, France

Prof. Shoei-Sheng Lee Taipei, Taiwan

Prof. Francisco Macias Cadiz, Spain

Prof. Anita Marsaioli Campinas, Brazil

Prof. Imre Mathe Szeged, Hungary

Prof. Joseph Michael Johannesburg, South Africa

Prof. Ermino Murano Trieste, Italy

Prof. Virinder Parmar Delhi, India

Prof. Luc Pieters Antwerp, Belgium

Prof. Om Prakash Manhattan, KS, USA

Prof. Peter Proksch Düsseldorf, Germany

Prof. Satyajit Sarker Wolverhampton, UK

Prof. Raffaele Riccio Salerno, Italy

Prof. Monique Simmonds Richmond, UK

Prof. Valentin Stonik Vladivostok, Russia

Prof. Hiromitsu Takayama Chiba, Japan

Prof. Karen Valant-Vetschera Vienna, Austria

Prof. Peter G. Waterman Lismore, Australia

Prof. Paul Wender Stanford, USA

HONORARY EDITOR

PROFESSOR GERALD BLUNDEN The School of Pharmacy & Biomedical Sciences,

University of Portsmouth, Portsmouth, PO1 2DT U.K.

[email protected]

Pancreatic Lipase Inhibitory Alkaloids of Murraya koenigii Leaves Rahul Birari, Somendu Kumar Roy, Anubha Singh and Kamlesh Kumar Bhutani*

Department of Natural Products, National Institute of Pharmaceutical Education and Research, (NIPER), S.A.S. Nagar, Punjab - 160 062, India [email protected] or [email protected] Received: March 15th, 2009; Accepted: July 27th, 2009

In the continuing search for newer pancreatic lipase inhibitors from plants, a total of 63 extracts from 21 different plants were screened to study their pancreatic lipase (PL) inhibitory activity in vitro. All three extracts (DCM, EtOAc and MeOH) of Murraya koenigii (L.) Spreng leaves (Rutaceae) exhibited antilipase activity greater than 80%. Further, bioactivity guided fractionation of the EtOAc extract led to the isolation of four alkaloids, namely mahanimbin, koenimbin, koenigicine and clausazoline-K, with IC50 values of 17.9 µM, 168.6 µM, 428.6 µM and <500 µM, respectively. This study reports for the first time the PL inhibitory potential of carbazole alkaloids from plants. Keywords: Murraya koenigii, Rutaceae, pancreatic lipase, mahanimbin, koenimbin, koenigicine, clausazoline-K, obesity. Physiologically, obesity is a disarray of energy balance and is primarily considered as a disorder of lipid metabolism [1]. Because excess fat consumption is widely thought to be one of the main causes of obesity, means of specifically inhibiting triglyceride (TG) digestion have been developed as a new approach to reducing fat absorption. A growing number of enzymes involved in lipid metabolic pathways are being identified and characterized; as such they represent a rich pool of potential therapeutic targets for obesity [2-4]. Pancreatic lipase (triacylgycerol acyl hydrolase), the principal lipolytic enzyme synthesized and secreted by the pancreas, plays a key role in the efficient digestion of triglycerides hydrolyzing 50-70% of dietary fats. Orlistat, one of the few drugs approved for antiobesity therapy, acts through inhibition of pancreatic lipase [5,6]. Many plants and their metabolites have earlier been reported to be inhibitors of pancreatic lipase and can form an important lead in the development of safe and effective antiobesity therapeutics. These inhibitors were recently reviewed by us [7,8]. In order to find newer pancreatic lipase (PL) inhibitors, 21 common medicinal plants were

selected, through literature survey (see Table 1), on the basis of their hyperlipidemic potential. These were screened for their antilipase activity in vitro. According to the literature survey, except for Alpinia officinarum [9,10] and Zinziber officinale [11], none of the other plants had been screened previously for antilipase activity. The models used for the screening of plants in earlier studies were different from those used in our study. Both these plants showed good activity in our assay. The lipase inhibitory activity of these extracts was calculated as percent inhibition. Table 1 records the inhibitory activity of the tested plants towards pancreatic lipase. Amongst the screened extracts, 21 showed stronger inhibition potential with percent inhibition values greater than 75%. DCM extracts of G. glabra, S. anacardium and E. prostrata; ethyl acetate extracts of G. glabra and M. koenigii; and methanol extracts of M. koenigii and S. anacardium exhibited antilipase activity greater than 90%, indicating their high potential as sources of natural pancreatic lipase inhibitors. The activity of these extracts was compared with the standard drug orlistat, which

NPC Natural Product Communications 2009 Vol. 4 No. 8

1089 - 1092

1090 Natural Product Communications Vol. 4 (8) 2009 Birari et al.

Table 1: Pancreatic lipase inhibitory activity of the screened plants at 1mg/mL.

Percent Inhibition ± SEM Sr. No.

Plant name (Family) Plant part used DCM extract Ethyl acetate extract Methanol extract

1 Acorus calamus L. (Acoraceae) AC Rhizome 41.8±2.5 30.9±2.9 Inactive 2 Alpinia officinarum Hance. (Zingiberaceae) AO Rhizome 86.7±0.5 72.6±0.6 Inactive 3 Caesalpinia bonducella L Roxb. (Caesalpiniaceae) CB Seeds Inactive 13.5±3.2 Inactive 4 Cassia tora L. (Caesalpiniaceae) CT Seeds 41.5±3.8 20.4±4.9 51.5±1.2 5 Eugenia jambolana Lam. (Myrtaceae) EJ Seeds 63.2±0.3 40.8±0.5 57.5±3.1 6 Mucuna pruriens DC. (Leguminosae) MP Seeds 62.1±1.4 44.7±2.0 74.8±1.3 7 Pterocarpus marsupium Roxb. (Leguminosae) PM Heart wood 89.1±0.4 85.2±0.5 79.4±1.9 8 Semecarpus anacardium L.f. (Anacardiaceae) SA Seeds 98.6±0.8 64.6±0.4 93.1±0.1 9 Tinospora cordifolia Miers (Menispermaceae) TC Stems 74.3±1.8 63.1±0.6 48.5±1.6 10 Hemidesmus indica (Asclepiadaceae) HI Seeds 51.2±3.7 4.1±1.9 61.7±0.4 11 Zingiber officinale Roscoe. (Zingiberaceae) ZO Rhizomes 84.6±0.8 67.6±0.6 13.2±3.4 12 Achyranthes aspera Wall. (Amaranthaceae) AA Whole plant 80.2±2.0 87.5±0.7 35.6±0.8 13 Aegle marmelos Correa. (Rutaceae) AM Leaves 80.1±0.8 79.6±0.7 46.9±1.8 14 Asparagus racemosus Willd. (Asparagaceae) AR Fresh roots 50.7±3.1 56.7±0.3 Inactive 15 Cinnamomum tamala Buch.-Ham. (Lauraceae) CiT Leaves 85.2±1.9 84.7±0.5 48.5±0.7 16 Eclipta prostrata L. (Asteraceae) EP Whole plant 91.7±0.9 86.5±0.5 Inactive 17 Ficus benghalensis L. (Moraceae) FB Bark 56.6±1.5 60.9±0.6 79.2±0.5 18 Momordica charantia Descourt. (Cucurbitaceae) MC Fruit powder 45.8±2.1 43.5±1.3 Inactive 19 Murraya koenigii (L.) Spreng. (Rutaceae) MK Leaves 84.2±1.2 95.4±0.3 92.2±0.6 20 Terminalia arjuna Wight & Arn. (Combretaceae) TA Bark 66.3±2.2 24.8±0.7 65.7±1.8 21 Glycyrrhiza glabra L. (Fabaceae) GG Roots 98.0±0.1 99.2±0.1 78.8±2.1

strongly inhibited pancreatic lipase with a calculated IC50 value of 0.015 µM ± 0.013. The inhibitory activity of the plant extracts was weaker than that of the standard drug orlistat. This was expected, as extracts are multi-constituent systems. These extracts can be further fractionated to find out the more potent PL inhibitory compounds. The results suggested that the DCM and ethyl acetate extracts are more active than the respective methanolic extracts. This can be partly explained by the constitution of these extracts. In general, these extracts are rich in flavonoids and terpenes and these types of constituents are expected to be responsible for the stronger activity of the extracts. M. koenigii, commonly known as ‘Curry Patta’, is traditionally used in India as a spice for its characteristic flavor and aroma [12]. The aromatic leaves are considered as a tonic, anthelmintic, analgesic, digestive, and appetizer, being widely used in Indian cookery for flavoring food stuffs. The green leaves have been used for the treatment of piles, inflammation, itching, fresh cuts, dysentery, vomiting, burses and dropsy. Aqueous extracts of M. koenigii have a strong hypolipidemic activity and have been indicated for the treatment of the mild form of diabetes, [13]. The aqueous extract of M. koenigii leaves contains a range of active

MKE-3 MKE-4

NH

H3CO

H3COO

NH

O

NH

H3CO

O

NH

CHO

H3CO

MKE-1 MKE-2

1

2345

6

7

81' 2'

3'

1"2"

3"4"

5"

6"

Figure 1: Structures of isolated alkaloids. pharmacological agents, including carbazole alkaloids, flavonoids and tannins [14,15]. In this present study, a phytochemical investigation was made of M. koenigii to find the responsible constituents for the observed activity of the extracts. Four known carbazole alkaloids mahanimbin (MKE 1), koenimbin (MKE 2), koenigicine (MKE 3) and clausazoline-K (MKE 4) were isolated and identified by comparison of their spectral data with those published in literature [16-21]. The isolated compounds were assayed for PL inhibition in vitro and the IC50 values were determined. The IC50 values were compared with that of orlistat. Mahanimbin, koenimbin, koenicine and clausazoline-K inhibited PL with IC50 values of 17.9 µM, 168.6 µM, 428.6 µM and <500 µM, respectively.

Pancreatic lipase inhibitors from Murraya koenigii Natural Product Communications Vol. 4 (8) 2009 1091

The higher inhibition by mahanimbin may be associated with its prenylated side chain. Thus prenylated carbazole alkaloids may form the natural template for synthesizing newer PL inhibitors. The study further indicates that the carbazole alkaloids of M. koenigii are major active constituents for the inhibition of PL and thus can be taken as a prototype for developing more potent PL inhibitors for the treatment of obesity. Further in vivo experiments are currently underway in our laboratory to ascertain the effects of M. koenigii alkaloids on lipid metabolism. Experimental

General: The 1H NMR and 13C NMR spectra were recorded on a Bruker DPX-300 instrument. Mass spectra were recorded on a Bruker daltonics Ultraflex MALDI-TOF/TOF instrument. Plants were extracted with a Dionex Accelerated Solvent Extractor. Scanning utilized a Labsystem multiscan microplate reader, while TLC profiling was done on a CAMAG HPTLC instrument. Lipase from porcine pancreas, type II and 4-nitrophenylpalmitate were purchased from Sigma Chemical Co., St. Louis, MO (USA). Solvents used in the extraction were of LR grade and were purchased from Rankem and CDH limited. Plant materials: Achyranthes aspera, Aegle marmelose, Asparagus racemosus, Cinnamomum tamala, Eclipta prostrata, Ficus bengalensis, Momordica charantia, M. koenigii, and Terminalia arjuna were collected from the institute campus. The other plant materials were bought from the local market of Chandigarh. The plants were authenticated by the qualified botanist in the medicinal plant garden of NIPER. A collection of voucher specimens is available at the NIPER nursery. Preparation of plant extracts: The respective plant materials were dried in an oven at 400 C and ground to a coarse powder. The powdered material (50 g) was sequentially extracted with n-hexane,

dichloromethane, ethyl acetate and methanol in an Accelerated Solvent Extractor (ASE). The n-hexane extract was discarded and the others were dried in a rotary evaporator and stored under refrigeration conditions. Isolation of alkaloids from M. koenigii: Four alkaloids (Figure 1) were isolated from the EtOAc extract of M. koenigii leaves. The extract (10 g) was loaded onto silica gel (# 60-120) and eluted with an increasing gradient of C6H6: CHCl3 and then CHCl3: CH3OH. Five fractions were obtained based on similar TLC patterns. Fraction 2 was purified by prep. TLC to give MKE-1 (94 mg), using C6H6: CHCl3 (3:2) as mobile phase. Fraction 6 was further re-chromatographed to yield MKE-2 (20 mg), MKE-3 (35 mg) and MKE-4 (40 mg), respectively. Screening methodology: For PL inhibitory activity, the assay procedure [22] was standardized. Lipase activity was measured using 4-nitrophenyl palmitate (PNPP) as a substrate. Porcine PL (Type II, crude) was dissolved in Tris-HCl buffer (pH 8.5) to give a concentration of 1mg/mL. The solution was stored at -20ºC. PNPP was dissolved in acetonitrile to give a stock solution of 10 mM. This was further diluted with ethanol (1:2 v/v) resulting in 3.33 mM PNPP. The solution was stored at -200C. Tris-HCl buffer 0.1 mM was prepared and the pH was adjusted to 8.5 by addition of conc. HCl. Extract solution were prepared by dissolving 1 mg of extract in 1mL of Tris-HCl buffer containing 50 µL of dimethyl sulfoxide. The final 200 μL reaction mixture contained 0.1 mg/mL of porcine PL, 0.167 mM PNPP and 0.25 mg/mL of extract/inhibitor in DMSO. The final volume was adjusted by Tris HCl buffer, pH 8.5. The reaction mixture was incubated at 370C. After incubation for 30 mins, the amount of 4-nitro phenol released by the lipase was measured at 405 nm in a Multiskan microtitre plate reader. All the experiments were performed in triplicate.

References [1] Strader CD, Hwa JJ, Van Heek M, Parker EM. (1998) Novel molecular targets for the treatment of obesity. Drug Discovery Today,

3, 250-256. [2] Foster-Schubert KE, Cummings DE. (2006) Emerging therapeutic strategies for obesity. Endocrine Reviews, 27, 779-793. [3] Shi Y, Burn P. (2004) Lipid metabolic enzymes: Emerging drug targets for the treatment of obesity. Nature Reviews: Drug

Discovery, 3, 695-710. [4] Melnikova I, Wages D. (2006) Antiobesity therapies. Nature Reviews: Drug Discovery, 5, 369-370. [5] Mukherjee M. (2003) Human digestive and metabolic lipases-a brief review. Journal of Molecular Catalysis B. Enzymatic, 22,

369-376. [6] Isler D, Moeglen C, Gains N, Meier MK. (1995) Effect of the lipase inhibitor orlistat and of dietary lipid on the absorption of

radiolabelled triolein, tri-γ-linolenin and tripalmitin in mice. British Journal of Nutrition, 73, 851-862.

1092 Natural Product Communications Vol. 4 (8) 2009 Birari et al.

[7] Birari RB, Bhutani KK. (2007) Pancreatic lipase inhibitors from natural sources: unexplored potential. Drug Discovery Today, 12, 879-889.

[8] Bhutani KK, Birari RB, Kapat K. (2007) Potential antiobesity and lipid lowering natural products: a review. Natural Product Communications, 2, 331-348

[9] Shin J, Han MJ, Kim D. (2002) 3-Methylethergalangin isolated from Alpinia officinarum inhibits pancreatic lipase. Biological and Pharmaceutical Bulletin, 26, 854-857.

[10] Shin J, Han MJ, Song M, Baek N, Kim D. (2004) 5-Hydroxy-7-(4’-hydroxy-3'-methoxyphenyl)-1-phenyl-3-heptanone: A pancreatic lipase inhibitor isolated from Alpinia officinarum. Biological and Pharmaceutical Bulletin, 27, 138-140.

[11] Han LK, Gong X, Kawano S, Saito M, Kimura Y, Okuda H. (2005) Antiobesity action of Zingiber officinale Roscoe. Yakugaku Zasshi, 125, 213-217.

[12] Chopra RN, Nayar SL, Chopra IC. (1996) Glossary of Indian Medicinal Plants. CSIR, New Delhi, p. 470. [13] Kesari AN, Kesari S, Singh SK, Gupta RK, Watal G. (2007) Studies on the glycemic and lipidemic effect of Murraya koenigii in

experimental animals. Journal of Ethnopharmacology, 112, 305-311. [14] Chakrabarty M, Nath A, Khasnobis S, Chakrabarty M, Konda Y, Harigaya Y, Komiyama K. (1997) Carbazole alkaloids from

Murraya koenigii. Phytochemistry, 46, 751-755. [15] (a) Wang YS, He HP, Shen YM, Hong X, Hao XJ. (2003) Two new carbazole alkaloids from Murraya koenigii. Journal of Natural

Products, 66, 416-418; (b) Mandal S, Nayak A, Banerjee SK, Banerji J, Banerji A. (2008) A new carbazole alkaloid from Murraya koenigii Spreng (Rutaceae). Natural Product Communications, 3, 1679-1682; (c) Chakraborty M, Saha S, Mukhapadhyay S. (2009) Murrayakoeninol- a new carbazole alkaloid from Murraya koenigii (Linn) Spreng. Natural Product Communications, 4, 355-358.

[16] Narasimhan NS, Paradkar MV, Chitguppi VP. (1968) Structures of mahanimbin and koenimbin. Phytochemistry, 53, 5501-5504. [17] Mukherjee M, Mukherjee S, Shaw AK, Ganguly SN (1983) Mukonicin a carbazole alkaloid from leaves of M. koenigii.

Phytochemistry, 22, 2328-2329. [18] Sharma RB, Verma RS, Kapil RS. (1980) Synthesis of koenigicine. Cellular and Molecular Life Sciences, 36, 815. [19] Chihiro I, Shinya K, Ohta H, Kajiura I, Furukawa H. (1997) Constituents of Clausena excavate. Isolation and structural elucidation

of new carbazole alkaloids. Chemical Pharmaceutical Bulletin, 45, 48-52. [20] Li WS, McChesney JD, El-Feraly FS. (1991) Carbazole alkaloids from Clausena lansium. Phytochemistry, 30, 343-346. [21] Krahl MP, Jager A, Krause T, Knolker HJ. (2006) First total synthesis of the 7-oxygenated carbazole alkaloids clauszoline-K, 3-

formyl-7-hydroxycarbazole, clausine M, clausine N and the anti-HIV active siamenol using a highly efficient palladium-catalyzed approach. Organic & Biomolecular Chemistry, 4, 3215-3219.

[22] Slanc P, Doljak B, Mlinaric A, Strukelj B. (2004) Screening of wood damaging fungi and macrofungi for inhibitors of pancreatic lipase. Phytotherapy Research, 18, 758-762.

Recovery and Purification of Nicotine from Waste Tobacco by Aqueous Two-Phase System/Reverse Extraction Jian-min Xing, Fen-fang Li and Jing Ping 1093

Structure Elucidation of Acylsucrose Derivatives from Atractylodes lanceae Rhizome and Atractylodes Rhizome Ken Tanaka and Atsutoshi Ina 1095

The Composition of the Essential Oil from Cyperus distans Rhizome Oladipupo A. Lawal and Adebola O. Oyedeji 1099

Essential Oil Composition and Enantiomeric Distribution of Fenchone and Camphor of Lavandula cariensis and L. stoechas subsp. stoechas grown in Greece Olga Tzakou, Ioannis Bazos and Artemios Yannitsaros 1103

Chemical Composition, Olfactory Evaluation and Antioxidant Effects of Essential Oil from Mentha x piperita Erich Schmidt, Stefanie Bail, Gerhard Buchbauer, Ivanka Stoilova, Teodora Atanasova, Albena Stoyanova, Albert Krastanov and Leopold Jirovetz 1107

Analysis of Essential Oils of Artemisia absinthium L. from Lithuania by CC, GC(RI), GC-MS and 13C NMR Asta Judzentiene, Félix Tomi and Joseph Casanova 1113

Chemical Composition and Antibacterial Activity of the Essential Oil of Cordia verbenacea from the Venezuelan Andes Gina Meccia, Luis B. Rojas, Judith Velasco, Tulia Díaz, Alfredo Usubillaga, Juan Carmona Arzola and Sulymar Ramos 1119

Composition and Antimicrobial Activity of the Leaf Essential oil of Litsea kostermansii from Taiwan Chen-Lung Ho, Eugene I-Chen Wang, Kuang-Ping Hsu , Pei-Yeh Lee and Yu-Chang Su 1123

Chemical Composition and Antimicrobial Activity of the Essential Oils of Leaves, Roots and Bark of Glossocalyx staudtii Huguette Agnaniet, Thomas Makani, Raphaël Bikanga, Louis Clément Obame, Jacques Lebibi and Chantal Menut 1127

Composition, Antioxidant and Antimicrobial Activities of the Essential Oil of Marrubium deserti Hocine Laouer, Benalia Yabrir, Amar Djeridane, Mohamed Yousfi, Nicolas Beldovini and Mebarka Lamamra 1133

Anti-Helicobacter pylori activity and Essential Oil Composition of Thymus caramanicus from Iran Fereshteh Eftekhar, Farahnaz Nariman, Morteza Yousefzadi, Javad Hadian and Samad Nejad Ebrahimi 1139

Differential Acetyl Cholinesterase Inhibition by Volatile Oils from two Specimens of Marlierea racemosa (Myrtaceae) Collected from Different Areas of the Atlantic Rain Forest Amanda Souza, Michelle C. Silva, Elaine M. Cardoso-Lopes, Inês Cordeiro, Marcos E. G. Sobral, Maria Cláudia M. Young and Paulo R. H. Moreno 1143

Factors Affecting Chemical Variability of Essential Oils: a Review of Recent Developments Andrea Barra 1147

Microbial Transformation of Sesquiterpenoids Haq N Bhatti, Muhammad Zubair, Nasir Rasool, Zahid Hassan and Viqar U Ahmad 1155

Natural Product Communications 2009

Volume 4, Number 8

Contents

Original Paper Page

A New Sesquiterpenoid Coumarin from Ferula assafoetida Anindita Ghosh, Avijit Banerji, Suvra Mandal and Julie Banerji 1023

Antiproliferative Activities of Five Chinese Medicinal Herbs and Active Compounds in Elephantopus scaber Miaoxian Su, Xia Wu, Hau Yin Chung, Yaolan Li and Wencai Ye 1025

Naturally Occurring Labdane Diterpene and Benzofuran from Globba pendula Maulidiani, Khozirah Shaari, Christian Paetz, Johnson Stanslas, Faridah Abas and Nordin Haji Lajis 1031

VCD Determination of the Absolute Configuration of Stypotriol Marcelo A. Muñoz, Carlos Areche, Aurelio San-Martín, Juana Rovirosa and Pedro Joseph-Nathan 1037

Bioactive Steroidal Sulfates from the Ambulakrums of the Pacific Starfish Lysastrosoma anthosticta Eleonora V. Levina, Anatoly I. Kalinovsky, Pavel S. Dmitrenok and Dmitry L. Aminin 1041

Role of Modifier in Microwave Assisted Extraction of Oleanolic Acid from Gymnema sylvestre: Application of Green Extraction Technology for Botanicals Vivekananda Mandal, Saikat Dewanjee and Subhash C. Mandal 1047

Phytochemical Characterization of an Adaptogenic Preparation from Rhodiola heterodonta Mary H. Grace, Gad G. Yousef, Anvar G. Kurmukov, Ilya Raskin and Mary Ann Lila 1053

Chemical Composition and Antimicrobial Activity of Wild Garlic Allium ursinum of Bulgarian Origin Antoaneta Ivanova, Bozhanka Mikhova, Hristo Najdenski, Iva Tsvetkova and Ivanka Kostova 1059

Evaluation of the Antioxidant and Antimicrobial Properties of in vitro Cultured Drosera intermedia Extracts Tomás Grevenstuk, Sandra Gonçalves, Sara Almeida, Natacha Coelho, Célia Quintas, Maria Nelma Gaspar and Anabela Romano 1063

Determination of Phytoestrogen Composition in Soybean Cultivars in Serbia Jelena Cvejić, Đorđe Malenčić, Vesna Tepavčević, Mihalj Poša and Jegor Miladinović 1069

Antilonomic Effects of Brazilian Brown Seaweed Extracts Thaisa Francielle Souza Domingos, Carla Carvalho, Laura de Andrade Moura, Valéria Laneuville Teixeira, Renato Crespo Pereira, Éverson Miguel Bianco, Wilton José Ferreira, Carlos José Brito Ramos, Ana Luiza Palhares de Miranda, Paulo Assis Melo, Jorge Almeida Guimarães and André Lopes Fuly 1075

Rapid Identification of two Species of Peucedanum by High-Performance Liquid Chromatography-Diode Array Detection-Electrospray Ionization Tandem Mass Spectrometry Yanyan Tao, Jianguang Luo, Yuanyuan Lu, Deran Xu, Zhiguo Hou and Lingyi Kong 1079

Aaptamine Alkaloids from the Vietnamese Sponge Aaptos sp. Larisa K. Shubina, Anatoly I. Kalinovsky, Sergey N. Fedorov, Oleg S. Radchenko, Vladimir A. Denisenko, Pavel S. Dmitrenok, Sergey A. Dyshlovoy, Vladimir B. Krasokhin and Valentin A. Stonik 1085

Pancreatic Lipase Inhibitory Alkaloids of Murraya koenigii Leaves Rahul Birari, Somendu Kumar Roy, Anubha Singh and Kamlesh Kumar Bhutani 1089

Continued inside back cover