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
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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