Dual inhibition of cyclooxygenase (COX) and lipoxygenase (LOX) pathways is promising approach in treatment of inflammatory diseases. Drugs able to block production of prostanoids together with leukotrienes should provide better anti-inflammatory properties and fewer side effects than non-steroidal anti-inflammatory drugs (NSAIDs) and selective COX-2 inhibitors [1,2]. Our previous studies revealed that some natural quinone compounds such as primin, alkannin, or shikonin are potent in vitro COX inhibitors [3]. In the current study, we tested 19 quinone compounds for 5-lipoxygenase (5-LOX) inhibition using in vitro assay according to [4] where neutrophile granulocytes with 5-LOX activity isolated from the human blood are incubated with arachidonic acid (substrate) and tested samples. After the incubation, the amount of leukotriene B4 (LTB4) is determined by commercial LTB4 EIA kit (Assay Designs). The most active quinone 5-LOX inhibitors were benzoquinone primin and naphthoquinone shikonin which decreased LTB4 production by 93 and 87% at 50µM concentration, respectively. Reference inhibitor zileuton reduced LTB4 production by 91% at the same concentration. Based on these preliminary results obtained in 5-LOX assay together with data from previous studies targeted on COX inhibition the plant quinones such as primin and shikonin should be considered for further studies aimed on their potential of dual COX/LOX inhibition.

Tonsipret® is a commercially available medicinal product for the treatment of sore throat and tonsillitis. The herbal extracts used in this preparation are hydro-alcoholic tinctures of dried, ripe fruits from Capsicum annuum L., of resin from Guaiacum officinale L., and of freshly collected roots from Phytolacca americana L.. The anti-inflammatory activity of the single extracts was evaluated in-vitro concerning inhibition of prostaglandin biosynthesis by cyclooxygenases (COX-1 and -2) [1], inhibition of leukotriene biosynthesis in human neutrophile granulocytes [1], inhibition of nitric oxide (NO) production in RAW 264.7 macrophages [2], and inhibition of Nuclear Factor kappa B1 (NF-κB1) and COX-2 expression in THP-1 cells [3]. Each herbal extract appeared to have an anti-inflammatory tendency in-vitro. The extract of Guaiacum officinale showed the strongest inhibitory effects on the assessed parameters. Its active constituents interfered with NF-κB1 expression, as well as with COX-2 expression, the enzymatic activity of inducible nitric oxide synthase (iNOS), and 5-lipoxygenase – genes containing an NF-κB binding site in their promoter regions. The other examined extracts also revealed a certain anti-inflammatory activity. The extract of Capsicum annuum exhibited a good inhibition on the enzymatic activity of COX-2, whereas the extract of Phytolacca americana showed a higher inhibitory activity on COX-2 expression.

The Chinese herbal formula Huang Lian Jie Du Tang, consisting of Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri, and Fructus Gardeniae, is used in China as a decoction to clear heat and to relieve toxicity. According to Western medicine it is used in the treatment of gastritis, hypertension, cerebrovascular diseases, liver diseases, against eczema, chronic inflammations of the intestine, as well as for psychiatric disorders like schizophrenia and depression. Previous investigations demonstrated antioxidant and anti-cancer activities of the mixture, as well as its abilities to inhibit the progression of arterios progression of arteriosclerosis and to lower plasma triglycerides [1,2,3,4,5,6]. clerosis and to lower plasma triglycerides [1,2,3,4,5,6]. The aim of our study was to compare the effects of the mixture and of the single herbs concerning their ability to inhibit leukotriene biosynthesis in human granulocytes and prostaglandin formation by COX-1 and COX-2. Decoctions of the mixture and of the single herbs were lyophilized and tested. Moreover, decoctions were fractionated using liquid-liquid chromatography with n-heptane, dichlormethane, ethylacetate and n-butanol. Leukotriene and prostaglandin biosynthesis inhibition assays were performed as previously described [7,8]. The investigation showed that Huang Lian Jie Du Tang and decoctions of the single herbs are inhibiting the formation of LTB4 in human granulocytes, as recently described [9], but have hardly any inhibitory effect on cyclooxygenases (Fig.1)

Different 2-alkyl-4(1H)-quinolones from the fruits of Euodia rutaecarpa Hook. f. & Thomson (Rutaceae) inhibited leukotriene biosynthesis in previous studies [1]. It was concluded that the characteristic lipophilic side chain might be essential for the observed activities. In order to substantiate this assumption, derivatives of 1-methyl-4(1H)-quinolones with variations of the side chain at C-2 differing in length and unsaturation were synthesised and tested for inhibition of leukotriene B4 formation in an in vitro assay using activated neutrophile granulocytes [2]. A saturated side chain as well as a trans double bond at C-1′ lead only to moderate inhibition at 50µM (15.1–28.3% inhibition of LTB4 formation) whereas compounds with a cis double bond in combination with chain lengths between C-10 and C-14 showed the highest activities (42.1–93.0% inhibition at 50µM). Increasing the chain length to C-20 significantly lowered the activity (9.8% inhibition; positive control zileuton 78.5% inhibition at 10µM). The most active compound revealed no cytotoxicity at 300µM even after 48h incubation time in MRC-5 cells (XTT assay) [3]. It can be concluded that the length and saturation of the lipophilic side chain at C-2 in 1-methyl-4(1H)-quinolones exert a significant influence on leukotriene biosynthesis in neutrophile granulocytes.

The decoction of the aerial parts of Centaurea sadleriana JANKA (Asteraceae), a plant native to Hungary, is traditionally used to treat the wounds of sheep in the Southern Great Plain region. Phytochemical and pharmacological studies on this plant have not been performed so far. Only we have recently confirmed the wound healing effect of the plant on rats [1]. The objective of the present work was the in vivo and in vitro investigation of the anti-inflammatory effect of C. sadleriana and the isolation and identification of its active compounds. The concentrated methanol extract of aerial parts of C. sadleriana was partitioned using n-hexane and chloroform. These two fractions were further fractionated via VLC and the anti-inflammatory effects of the fractions were studied by in vitro (COX-1, COX-2 and LTB4 formation inhibitory activity) and in vivo (intraperitoneal and oral administration to rats) methods. Some of the fractions gained from the n-hexane extract possessed marked in vitro (70–85% LTB4 formation inhibition, 59–83% COX-1 inhibition, 80–92% COX-2 inhibition at a concentration of 50µg/ml) and in vivo anti-inflammatory effects (25–50% oedema volume reduction). Chromatographic purification (VLC, MPLC, HPLC, preparative TLC, CPC and gel filtration) of the active fractions resulted in the isolation of flavonoids, triterpenoids and lignans. Our present study confirmed the marked anti-inflammatory effect of C. sadleriana which may play role in the ethnomedicinal application of the plant.

The Chinese herbal mixture Huang Lian Jie Du Tang is used in traditional Chinese medicine to clear heat and relieve toxicity. According to Western medical terminology it is used against eczema and skin diseases, chronic inflammatory intestinal diseases such as Morbus Crohn or Colitis ulcerosa, as well as against psychiatric disorders like schizophrenia and depression. Recent studies on the mixture showed that it is influencing LTB4 formation and NO production [1]. The aim of this study was to determine the inhibitory activity of the decoctions of the mixture and of the single herbs on NO production in RAW 264.7 macrophages as well as their influence on cell viability and proliferation using an XTT assay. The results of the mixture were compared with the activity of the single herbs. In addition, the decoctions were fractionated using liquid-liquid extraction with n-heptane, dichloromethane, ethyl acetate and n-butanol, and the gained fractions were also tested. The study showed that only the dichloromethane fraction of the decoction of Radix Scutellariae was able to inhibit NO production (Fig.1). The XTT assay (performed with CCRF-CEM leukemia cells) demonstrated no or only little influence of the decoctions and fractions on cell proliferation and cell viability. Further investigations are in progress to prove whether the inhibition of NO production is due to baicalein which is a major compound of Radix Scutellariae, or whether also other constituents contribute to the effect.

Aconite roots are known to be very toxic due to diterpene ester alkaloids [1]. Therefore, in Chinese medicine they are only used after processing. Hydrolysis of the ester groups decreases toxicity [2]. Nevertheless, several cases of poisoning by unprocessed or improperly processed aconite roots have been reported [3]. Recently we suggested a specific sample preparation method and HPLC assay for the analysis of toxic aconite alkaloids [4]. Now, an improved method with an even better resolution is presented. Using this method, we have determined the content of mesaconitine, aconitine and hypaconitine in 30 commercial samples of processed aconite roots. In most of the samples, toxic aconite alkaloids were not detectable, or only traces were found. However, in four samples we could detect more than 0.04% of hypaconitine and mesaconitine, the highest with a content of 0.16%. Therefore, this method is suggested as a purity test for the European Pharmacopoeia. Acute toxicity of batches high in hypaconitine and mesaconitine was also confirmed in CFLP mice. In the aconite monograph of the German Homeopathic Pharmacopoeia, alkaloids are determined by a titration method. We compared the results of HPLC analysis of toxic alkaloids (mesaconitine, aconitine and hypaconitine) with the results obtained by the titration method, and found no correlation. Samples which were lacking mesaconitine, aconitine and hypaconitine, still contained up to 0.2% alkaloids determined by titration. Therefore, titration of alkaloids is not appropriate as an assay for toxic alkaloids in aconite roots.

Imupret® is an alcoholic-aqueous extract of seven different herbal drugs: Radix Althaeae, Flores Chamomillae, Herba Equiseti, Folia Juglandis, Herba Millefolii, Cortex Quercus, and Herba Taraxaci. It is used for the treatment of recurrent infections of the respiratory tract, especially tonsillitis. The aim of the study was the assessment of the influence of the herbal extract combination Imupret® compared to selected single herbal extracts on its anti-inflammatory activity in vitro. Therefore pharmacological effects of Imupret® and its herbal components on arachidonic acid metabolism were investigated. The anti-inflammatory activity was measured by the inhibition of prostaglandin biosynthesis by cyclooxygenases (COX-1 and -2) [1] and the inhibition of leukotriene biosynthesis in human neutrophile granulocytes [2]. The inhibitory activity was calculated by the produced amount of prostaglandin E2 (PGE2) and leukotriene B4 (LTB4), respectively. This study compares different anti-inflammatory pharmacological effects in order to reveal whether there is an increase or decrease of enzyme inhibition caused by additional synergistic effects as a result of a multiple extract preparation.

Previously we reported the semisynthesis of 9 aconitine-derived lipo-alkaloids transesterified with fatty acids differing in the number of carbon atoms and double bonds in their chains [1]. When these compounds were tested for COX-1, COX-2 and LTB4 formation inhibitory activities it was found that 14-benzoylaconine-8-O-eicosapentaenoate exhibited notable activities through all three in vitro anti-inflammatory test systems (inhibition (%): COX-1: 54.51, COX-2: 66.07, LTB4: 45.96, at 50µM concentration). Although no data exist on the natural occurrence of aconitine derived lipo-alkaloids containing eicosanoic acid analogues at C-8, the prospective of further charting the possible structure-activity relationships of lipo-alkaloids is reinforced by papers dealing with the importance and natural roles of different unsaturated eicosanoic acid derivatives in the process of inflammation [2]. The present paper reports the semisynthesis of 7 eicosanoate analogues of aconitine-derived lipo-alkaloids, prepared according to the modified method of Bai et al [3]. In the reactions, aconitine was transesterified by eicosanoic, 11-eicosenoic, 8,11,14-eicosatrienoic, 11,14,17-eicosatrienoic, 8,11,14,17-eicosatetraenoic and 5,8,11,14,17-eicosapentaenoic acids resulting the corresponding 14-benzoylaconin-8-O-esters and pyroaconitine. The reaction mixtures were purified of necessity by gelfiltration, preparative TLC and centrifugal planar chromatography. Purity of the obtained lipo-alkaloids was proved with the aid of NMR spectroscopy. The COX-1, COX-2 and LTB4 formation inhibitory activities of this eicosanoate lipo-alkaloid series were also investigated. It was observed that the 14-benzoylaconine-8-O-eicosapentaenoate has the highest activities (inhibition (%): COX-1: 82.84, COX-2: 33.67, at 50µM concentration) between the eicosanoate analogues.

Processed aconite drugs are widely used in Eastern medicine as painkillers and antirheumatic agents. It is known that the traditional processing of aconite drugs increases the amount of lipo-alkaloids. In order to obtain information about the pharmacological potential of these compounds, semisynthesis of 9 aconitine-derived lipo-alkaloids was carried out and their COX-1, COX-2 and LTB(4) formation inhibitory activities were investigated. It was found that compounds esterified with unsaturated fatty acids demonstrated significant COX-2 inhibitory effects, while in the COX-1 assay only 14-benzoylaconine-8-O-eicosapentaenoate exerted remarkable activity. The inhibition of LTB(4) formation was pronounced in cases of long chain fatty acid derivatives.

Focus on Alternative and Complementary TherapiesVolume 12, Issue s1 p. 15-15 Rosa canina pseudfructus sine fructibus: a promising antirheumatic S Chrubasik, S Chrubasik Department of Forensic Medicine, University of Freiburg, Albertstr. 9, Neumarkt, GermanySearch for more papers by this authorC Chrubasik, C Chrubasik Department of Forensic Medicine, University of Freiburg, Albertstr. 9, Neumarkt, GermanySearch for more papers by this authorL Wiesner, L Wiesner Department of Forensic Medicine, University of Freiburg, Albertstr. 9, Neumarkt, GermanySearch for more papers by this authorU Widowitz, U Widowitz Institute of Pharmaceutical Sciences, Universitätsplatz 4, 8010 Graz, Austria, E-mail: sigrun.chrubasik@klinikum.uni-freiburg.deSearch for more papers by this authorEM Wenzig, EM Wenzig Institute of Pharmaceutical Sciences, Universitätsplatz 4, 8010 Graz, Austria, E-mail: sigrun.chrubasik@klinikum.uni-freiburg.deSearch for more papers by this authorR Bauer, R Bauer Institute of Pharmaceutical Sciences, Universitätsplatz 4, 8010 Graz, Austria, E-mail: sigrun.chrubasik@klinikum.uni-freiburg.deSearch for more papers by this author S Chrubasik, S Chrubasik Department of Forensic Medicine, University of Freiburg, Albertstr. 9, Neumarkt, GermanySearch for more papers by this authorC Chrubasik, C Chrubasik Department of Forensic Medicine, University of Freiburg, Albertstr. 9, Neumarkt, GermanySearch for more papers by this authorL Wiesner, L Wiesner Department of Forensic Medicine, University of Freiburg, Albertstr. 9, Neumarkt, GermanySearch for more papers by this authorU Widowitz, U Widowitz Institute of Pharmaceutical Sciences, Universitätsplatz 4, 8010 Graz, Austria, E-mail: sigrun.chrubasik@klinikum.uni-freiburg.deSearch for more papers by this authorEM Wenzig, EM Wenzig Institute of Pharmaceutical Sciences, Universitätsplatz 4, 8010 Graz, Austria, E-mail: sigrun.chrubasik@klinikum.uni-freiburg.deSearch for more papers by this authorR Bauer, R Bauer Institute of Pharmaceutical Sciences, Universitätsplatz 4, 8010 Graz, Austria, E-mail: sigrun.chrubasik@klinikum.uni-freiburg.deSearch for more papers by this author First published: 14 June 2010 https://doi.org/10.1111/j.2042-7166.2007.tb05863.xRead the full textAbout ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume12, Issues1December 2007Pages 15-15 RelatedInformation

The aerial parts of Chinese motherwort (Leonurus japonicus Houtt) (Yimucao) are commonly used in traditional Chinese medicine (TCM) to regulate menses and to treat associated conditions. As the herb is more and more used in Europe, it has been started to cultivate the plant in Germany [1]. Since bitterness is an important sensory feature of the plant, the aim of the investigation was to isolate and identify the chemical constituents responsible for the bitter taste of Yimucao. It was found that the bitter principle can be extracted by n-hexane. Therefore, the n-hexane extract of Herba Leonuri has been fractionated using Fast Centrifugal Partition Chromatography (FCPC).This led to a fraction containing a mixture of two compounds which seemed to be responsible for the bitter taste of Yimucao. Structure elucidation by MS and NMR spectroscopy led to two new epimeric diterpenoid compounds 1 and 2 (exact mass 392,2), which are related to furanic labdane type lactones previously found in Leonurus japonicus, Leonurus cardiaca, Leonotis leonurus and Marrubium vulgare [2,3].

Echinacea is among the most popular medicinal plants today and has a long history of use for the treatment of the common cold, upper respiratory infections and inflammatory diseases. Besides Echinacea purpurea and Echinacea angustifolia, also the roots of Echinacea pallida are used medicinally. Polyacetylenes and polyenes are the major lipophilic constituents of Echinacea pallida root extracts. They are natural compounds known for their antifungal and antibacterial activity, and have enzyme inhibitory effects [1]. There is some evidence that they might also exhibit antiallergic as well as anti-inflammatory activities, and recently cytotoxic effects have been found [2].

The aim of the present study was to compare powdered rose hip with and without fruits (Rosae pseudofructus cum/sine fructibus, Rosa canina L., Rosaceae) with regard to their phytochemical profile and their in vitro anti-inflammatory and radical-scavenging properties. The two powders were subsequently extracted with solvents of increasing polarity and tested for inhibition of cyclooxygenase (COX-1, COX-2) and of 5-LOX-mediated leukotriene B(4) (LTB(4)) formation as well as for DPPH-radical-scavenging capacity. While the water and methanol extracts were inactive in the COX-1, COX-2 and LTB(4) inhibition assays, the n-hexane and the dichloromethane extracts inhibited all three enzymes. In the active extracts, the triterpenoic acids ursolic acid, oleanolic acid and betulinic acid were identified, although only in minute amounts. Furthermore, oleic, linoleic and alpha-linolenic acid were identified apart from several saturated fatty acids. Even though unsaturated fatty acids are known to be good inhibitors of COX-1, COX-2 and LT formation, no clear correlation between their concentration in the extracts and their activity was found. We suggest that other, yet unidentified, lipophilic constituents might play a more important role for the observed in vitro inhibitory activity on arachidonic acid metabolism. Some of the extracts also showed considerable DPPH radical scavenging activity, the methanolic extracts being most potent. The radical scavenging activity of the extracts correlated very well with their total phenolic content, while ascorbic acid contributes only little to the radical-scavenging activity due to its low concentration present in the extracts. In summary, extracts derived from powdered rose hip without fruits were more effective in all assays carried out compared with extracts derived from powdered rose hip with fruits.

Rose hip with or without fruits (Rosae pseudofructus cum/sine fructibus, Rosa canina L., Rosaceae) is traditionally used for the prevention and therapy of common cold and other infections, as a diuretic agent, for the treatment of gout and rheumatic diseases and as a vitamin C source. For none of these indications clinical evidence of effectiveness has been demonstrated except for osteoarthritis: A proprietary rose hip and seed powder had moderate evidence of effectiveness in alleviating osteoarthritic complaints [1]. Little is known about the rose hip mechanism of action and active constituents. Aim of this study was to get information on the constituents responsible for the anti-inflammatory potential of rose hip peel.