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Planta Med 2001; 67(5): 411-416
DOI: 10.1055/s-2001-15805
Original Paper
Pharmacology
© Georg Thieme Verlag Stuttgart · New York

Identification and Isolation of the Cyclooxygenase-2 Inhibitory Principle in Isatis tinctoria

Henning Danz1 , Stefka Stoyanova1 , Petra Wippich1 , Axel Brattström2 , Matthias Hamburger1,*
  • 1 Institute of Pharmacy, Friedrich-Schiller-University Jena, Germany
  • 2 Zeller AG, Romanshorn, Switzerland
Further Information

Publication History

July 4, 2000

October 22, 2000

Publication Date:
31 December 2001 (online)

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Abstract

Various extracts prepared from the traditional dye and medicinal plant Isatis tinctoria L. were submitted to a broad in vitro screening against 16 anti-inflammatory targets. Dichloromethane (DCM) extracts from dried leaves showed a marked cyclooxygenase (COX) inhibitory activity with a preferential effect on COX-2 catalysed prostaglandin synthesis. A supercritical fluid extraction (SFE) procedure employing CO2-modifier mixtures was developed by which the bioactivity profile and chromatographic fingerprint of the DCM extract could be reproduced. High-resolution activity directed on-line identification of the COX-2 inhibitory principle, using a combination of LC-DAD-MS with a microtitre-based bioassay, led to the identification of tryptanthrin (1) as the constituent responsible for essentially all COX-2 inhibitory activity in the crude extract. Following on-line identification, 1 was isolated at preparative scale and its structure confirmed by comparison with synthetic tryptanthrin. In an assay with lipopolysaccharide stimulated Mono Mac 6 cells, tryptanthrin (1) was of comparable potency (IC50 = 64 nM) than the preferential COX-2 inhibitors nimesulide (IC50 = 39 nM) and NS 398 (IC50 = 2 nM). The SFE extract and 1 showed no cytotoxicity in Mono Mac 6 and RAW 264.7 cells when tested at 100 μg/ml and 10 μM, respectively.

Key words

Isatis tinctoria - Brassicaceae - tryptanthrin - anti-inflammatory activity - cyclooxygenase-2 inhibition - on-line spectroscopy - Mono Mac 6 cells

References

  • 1 Blumenthal M. The complete German Commission E monographs. Austin (Texas); American Botanical Council 1998
  • 2 Ernst E, Chrubasik S. Phyto-anti-inflammatories. A sytematic review of randomized, placebo-controlled, double-blind trials.  Rheumatic Diseases Clinic North America. 2000;  26 13-27
    PubMedSearch in Google Scholar
  • 3 Hurry J B. The woad plant and its dye. London; Oxford University Press 1930: 249-56
  • 4 Index Kewensis 2.0. London; Oxford University Press 1997
  • 5 Zhu Y. Chinese Materia Medica. Amsterdam; Harwood Academic 1998: 183-9
  • 6 Dornberger K, Lich H. Screening nach antimikrobiell sowie potentiell cancerostatisch wirksamen Pflanzeninhaltsstoffen.  Pharmazie. 1982;  37 215-21
    PubMedSearch in Google Scholar
  • 7 Seifert K, Unger W. Insecticidal and fungicidal compounds from Isatis tinctoria .  Z. Naturforsch.. 1994;  49c 44-8
    PubMedSearch in Google Scholar
  • 8 Diel F, Well S, Ficht F, Paetzold M. Tea of Isatis tinctoria (woad) responses by allergic patients in vivo and in vitro .  Aktuelle Ernährungsmedizin. 1992;  17 34-6
    PubMedSearch in Google Scholar
  • 9 You S. Anti-blood platelet aggregation constituents in the root of Isatis tinctoria L.  Zhongyao Tongbao. 1988;  13 31-2
    PubMedSearch in Google Scholar
  • 10 Honda G, Tosirisuk V, Tabata M. Isolation of an antidermatophytic, tryptanthrin, from indigo plants, Polygonum tinctorium and Isatis tinctoria .  Planta Medica. 1980;  38 275-6
    PubMedSearch in Google Scholar
  • 11 Wu X Y, Qin G W, Cheung K K, Cheng K F. New alkaloids from Isatis indigotica .  Tetrahedron. 1997;  53 13323-8
    CrossrefPubMedSearch in Google Scholar
  • 12 Hartleb I, Seifert K. A novel anthranilic acid derivative from Isatis tinctoria .  Planta Medica. 1994;  60 578-9
    PubMedSearch in Google Scholar
  • 13 Hartleb I, Seifert K. Acid constituents from Isatis tinctoria .  Planta Medica. 1995;  61 95-6
    PubMedSearch in Google Scholar
  • 14 Vogel H G, Vogel W H (eds.). Drug Discovery and Evaluation - Pharmacological Assays. Berlin; Springer 1997: 395-401
  • 15 Berg J, Christoph T, Widerna M, Bodenteich A. Isoenzyme-specific cyclooxygenase inhibitors: A whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6.  Journal of Pharmacological and Toxicological Methods. 1997;  37 179-86
    CrossrefPubMedSearch in Google Scholar
  • 16 Borenfreund E, Puerner J A. Toxicity determined in vitro by morphological alternations and neutral red absorption.  Toxicology Letters. 1985;  24 119-24
    CrossrefPubMedSearch in Google Scholar
  • 17 George V, Koshy A S, Singh O V, Nayar M NS, Pushpangadan P. Tryptanthrin from Wrightia tinctoria .  Fitoterapia. 1996;  67 53-4
    PubMedSearch in Google Scholar
  • 18 Friedländer P, Roschdestwensky N. Über ein Oxidationsprodukt des Indigblaus.  Chemische Berichte. 1915;  48 1841-7
    CrossrefPubMedSearch in Google Scholar
  • 19 Taylor L T. Supercritical Fluid Extraction. New York; Wiley 1996: 1-6
  • 20 Hamburger M, Hostettmann K. Bioactivity in plants: The link between phytochemistry and medicine.  Phytochemistry. 1991;  30 3864-74
    CrossrefPubMedSearch in Google Scholar
  • 21 Liu J H, Zschocke S, Reiniger E, Bauer R. Inhibitory effects of Angelica pubescens f. biserrata on 5-lipoxygenase and cyclooygenase.  Planta Medica. 1998;  64 525-9
    PubMedSearch in Google Scholar
  • 22 Kimoto T, Yamamoto Y, Hino K, Koya S, Aga H, Hashimoto T, Hanaya T et al. Cytotoxic effects of substances in indigo plant (Polygonum tinctorium Lour.) on malignant tumour cells.  Natural Medicines. 1999;  53 72-9
    PubMedSearch in Google Scholar
  • 23 Mitscher L, Baker W. Tuberculosis: A search for novel therapy starting with natural products.  Medicinal Research Reviews. 1998;  18 363-74
    CrossrefPubMedSearch in Google Scholar
  • 24 Talley J J. Selective inhibitors of cyclooxygenase-2 (COX-2). In: King FD, Oxford AW, editors Progress in Medicinal Chemistry. Amsterdam; Elsevier 1999: 201-34
    Search in Google Scholar
  • 25 Moon C, Murakami M, Kudo I, Son K H, Kim H P, Kang S S, Chang H W. A new class of COX-2 inhibitor, rutaecarpine from Evodia rutaecarpa .  Inflammation Research. 1999;  48 621-5
    PubMedSearch in Google Scholar
  • 26 Wang G J, Wu X C, Chen C F, Lin L C, Huang Y T, Shan J et al. Vasorelaxing action of rutaecarpine: Effects of rutaecarpine on calcium channel activities in vascular endothelial and smooth muscle cells.  Journal of Pharmacology and Experimental Therapeutics. 1999;  289 1237-44
    PubMedSearch in Google Scholar
  • 27 Sheu J R, Kan Y C, Hung W C, Su C H, Lin C H, Lee Y M et al. The antiplatelet activity of rutaecarpine, an alkaloid isolated from Evodia rutaecarpa, is mediated through inhibition of phospholipase C.  Thrombosis Research. 1998;  92 53-64
    CrossrefPubMedSearch in Google Scholar
  • 28 Abad T, McNaughton-Smith G, Fletcher W Q, Echeverri F, Diaz-Penate R, Tabraue C et al. Isolation of (S)-(+)-naproxene from Musa acuminata. Inhibitory effect of naproxene and its 7-methoxy isomer on constitutive COX-1 and inducible COX-2.  Planta Medica. 2000;  66 471-3
    Thieme ConnectPubMedSearch in Google Scholar
  • 29 Ishihara T, Kohno K, Ushio S, Iwaki K, Ikeda M, Kurimoto M. Tryptanthrin inhibits nitric oxide and prostaglandin E2 synthesis by murine macrophages.  European Journal of Pharmacology. 2000;  407 197-204
    CrossrefPubMedSearch in Google Scholar
  • 30 Hamburger M, Danz H. Arzneimittel zur Hemmung von NF-κB. European Patent Application 99810750.2;. filed Aug. 20, 1999
    Search in Google Scholar
  • 31 Danz H, Stoyanova S, Hamburger M, Brattström A. Identification and isolation of the COX-2 inhibitory principle in Isatis tinctoria L. (Brassicaceae).  Archiv der Pharmazie. 2000;  333 (Suppl. 1) 11
    PubMedSearch in Google Scholar

Prof. Dr. Matthias Hamburger

Institut für Pharmazie

Friedrich-Schiller-Universität Jena

Semmelweisstraße 10

07743 Jena

Germany

Email: B7HAMA@rz.uni-jena.de

Fax: ++49 3641 949842