Bioactive flavonoids of Tanacetum parthenium revisited

Bio-guided fractionation of an extract from Tanacetum parthenium showing activity as mitotic blocker allowed the isolation and identification of santin 3, jaceidin 2 and centaureidin 1. The latter two closely related flavonols, which, to the best of our knowledge, are isolated here together for the first time, form a mixture difficult to resolve and which is probably the reason for the confusion in the literature regarding their occurrence. Centaureidin 1 had an IC50 of 1 microM while jaceidin 2 and santin 3 were 200 times less active.     

Sesquiterpene lactones and other constituents from Tanacetum parthenium

A reinvestigation of the aerial parts of Tanacetum parthenium afforded in addition to known compounds three pinene derivatives, two spiroketal enol ether polyines, four germacranolides; and six guaianolides, two of them being endoperoxides, and two secoguaianolides. The structures were elucidated by high field ¹H NMR spectroscopy, mass spectroscopy and a few chemical transformations. The biogenetic relationships are discussed briefly.     

Metabolic conversion of dietary flavonoids alters their anti-inflammatory and antioxidant properties

The notion that dietary flavonoids exert beneficial health effects in humans is often based on in vitro studies using the glycoside or aglycone forms of these flavonoids. However, flavonoids are extensively metabolized in humans, resulting in the formation of glucuronide, methyl, and sulfate derivatives, which may have different properties than their parent compounds. The goal of this study was to investigate whether different chemical modifications of the same flavonoid molecule affect its biological and antioxidant activities. Hence, we studied the anti-inflammatory effects of several major human metabolites of quercetin and (-)-epigallocatechin-3-O-gallate (EGCG) by assessing their inhibitory effects on tumor necrosis factor α (TNFα)-induced protein expression of cellular adhesion molecules in human aortic endothelial cells (HAEC). HAEC were incubated with 1-30 μM quercetin, 3'- or 4'-O-methyl-quercetin, quercetin-3-O-glucuronide, and quercetin-3'-O-sulfate or 20-100 μM EGCG, 4'-O-methyl-EGCG, and 4',4'-di-O-methyl-EGCG, prior to coincubation with 100 U/ml of TNFα. 3'-O-Methyl-quercetin, 4'-O-methyl-quercetin, and their parent aglycone compound, quercetin, all effectively inhibited expression of intercellular adhesion molecule-1 (ICAM-1) with IC(50) values (concentration required for 50% inhibition) of 8.0, 5.0, and 4.4 μM, respectively; E-selectin expression was suppressed to a somewhat lesser but still significant degree by all three compounds, whereas vascular cell adhesion molecule-1 (VCAM-1) was not affected. In contrast, quercetin-3-O-glucuronide (20-100 μM), quercetin-3'-O-sulfate (10-30 μM), and phenolic acid metabolites of quercetin (20-100 μM) did not inhibit adhesion molecule expression. 4',4'-Di-O-methyl-EGCG selectively inhibited ICAM-1 expression with an IC(50) value of 94 μM, whereas EGCG (20-60 μM) and 4'-O-methyl-EGCG (20-100 μM) had no effect. The inhibitory effects of 3'-O-methyl-quercetin and 4',4'-di-O-methyl-EGCG on adhesion molecule expression were not related either to inhibition of NF-κB activation or to their antioxidant reducing capacity. Our data indicate that flavonoid metabolites have different biological and antioxidant properties than their parent compounds, and suggest that data from in vitro studies using nonmetabolites of flavonoids are of limited relevance in vivo.     

The stereospecificity of alcohol oxidases from Tanacetum vulgare and Candida boidinii.

The stereospecificity of the oxidation of trans-hex-2-en-1-ol and ethanol was determined for alcohol oxidases from $\text{Tanacetum vulgare}$ leaves and from the yeast $\text{Candida boidinii}$. It was found that both enzymes remove the pro-1R hydrogen. The enzyme from $\text{Candida boidinii}$ also removes the pro-1S hydrogen but to a much lesser degree.     

The inflorescence and leaf essential oils of Tanacetum vulgare L. var. vulgare growing wild in Lithuania

Forty samples of inflorescences and leaves of wild Tanacetum vulgare L. var. vulgare were collected in 20 habitats from Lithuania. The essential oils were analyzed by GC and GC/MS. The 57 identified compounds in the oils made up 80.7–99.6%. According to the cluster analysis the volatile oils were divided into four groups with 1,8-cineole (23.6–46.3%, 11 oils), trans-thujone (35.7–78.4%, 6 samples), camphor (19.8–61.8%, 17 oils) and myrtenol (13.1–24.9%, 6 samples) as main constituents. The inflorescences and leaves of tansy plants formed the oils with the same dominating constituent in 15 of the 20 habitats investigated. The leaves in five localities produced oils of the 1,8-cineole chemotype, while the inflorescences biosynthesized oils of the camphor type in three habitats and of the myrtenol type in two habitats. Amounts of the 1,8-cineole in all leaf oils were greater than that in inflorescence oils of the plants from the same locality. An opposite correlation was determined for camphor, myrtenol, cis- and trans-thujone. The myrtenol chemotype was not noticed earlier in the essential oils of T. vulgare.     

Substrate specificity of monoterpenol dehydrogenases from Foeniculum vulgare and Tanacetum vulgare

Soluble enzyme preparations from leaves of Foeniculum vulgare catalysed the pyridine nucleotide-dependent dehydrogenation of l-endo-fenchol to d-fenchone, while similar preparations from Tanacetum vulgare catalysed the pyridine nucleotide-dependent dehydrogenation of d-3-thujanol to d-3-thujone. The monoterpenol dehydrogenases were separated from alcohol (ethanol) dehydrogenase by gel-permeation chromatography, and the MW, pH optimum, cofactor preference and other general properties of these enzymes were examined. Specificity studies indicated that only a narrow range of monoterpenols, related to those structural classes produced in vivo, were oxidized by these dehydrogenases.     

Germacranolides and an alkyl glucoside from Tanacetum vulgare

The aerial parts of Tanacetum vulgare afforded two new germacranolides and an n-decyl glucoside. The structures have been elucidated by spectroscopic methods and chemical transformations.     

Isolation and General Characterization of Polysaccharides from Tansy Tanacetum vulgare L.

Using extraction with 0.75% aqueous ammonium oxalate, the following polysaccharide fractions were isolated: tanacetans TVF, TVS, and TVR from floscules, sprouts, and roots, respectively, of Tanacetum vulgare L., spread throughout the European North of Russia. The sugar chain of tanacetan TVF consists of D-galacturonic acid (61.4%), arabinose (14.7%), galactose (10.2%), and rhamnose (3.7%) as the main constituents as well as xylose, glucose, mannose, apiose, and 2-O-methylxylose in trace amounts. Tanacetans TVS and TVR were shown to differ in the sugar quantitative composition. They contain 67 and 28% galacturonic acid, respectively. A partial acid hydrolysis of the tanacetan TVF gave a polysaccharide fragment TVF1, -1,4-D-galacturonan (GalA 98.2%). Digestion with pectinase (-1,4-D-polygalacturonase) resulted in fragment TVF3, containing residues of arabinose (27.1%) and galactose (17.3%). NMR spectroscopy allowed detection of the terminal residues of -Araf and -Galp as well as of the residues of -Arap substituted in 3,5- and 5-positions. Thus, tanacetan TVF was proved to be a pectic polysaccharide.     

Spatial and developmental expression of key genes of terpene biosynthesis in Tanacetum parthenium

Feverfew (Tanacetum parthenium) is a medicinal plant belonging to the Asteraceae family. To improve understanding terpene metabolism in feverfew, the relative gene expression of four key genes coding 3-hydroxy-3-methylglutarylcoenzyme A reductase (HMGR) and germacrene A synthase (GAS) from the mevalonic acid pathway (MVA), as well as 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) and hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) from the methyl erythritol phosphate pathway (MEP), were examined. Target organs and tissues included young leaves (not fully expanded), mature leaves (fully expanded), flowers, stems, roots, and glandular trichomes. HMGR, DXR, and HDR were isolated and sequenced for the first time in feverfew. Real-time quantitative PCR analysis revealed differential expression of these genes in feverfew tissues and developmental stages.     

Biosynthesis of geraniol and nerol in cell-free extracts of Tanacetum vulgare

Cell-free extracts from leaves of Tanacetum vulgare synthesised geraniol and nerol (3,7-dimethylocta-trans-2-ene-1-ol and its cis isomer) in up to 11·9 and 2·4% total yields from IPP-[4-¹⁴C] and MVA-[2-¹⁴C] respectively. Optimum preparations were obtained from plant material just before the onset of flowering. The ratio of the monoterpenols varied 28-fold for different preparations under conditions where these products or their phosphate esters were not interconverted. Similar extracts incorporated α-terpineol-[¹⁴C] and terpinen-4-ol-[¹⁴C] (p-menth-1-en-8- and -4-ol respectively) in 0·05 to 2·2% yields into a compound tentatively identified as isothujone (trans-thujan-3-one), and preparations from flowerheads converted IPP-[4-¹⁴C] in 2·7% yield into geranyl and neryl β-d-glucosides. Inhibitors of IPP-isomerase had little effect on the incorporation of IPP into the monoterpenols in cell-free systems from which endogenous compounds of low molecular-weight had been removed. The inference that a pool of protein-bonded DMAPP or its biogenetic equivalent was present was supported by the demonstration that geraniol and nerol biosynthesised in the absence of the inhibitors were predominantly (65 to 100%) labelled in the moiety derived from IPP.     

Colonization of Tanacetum vulgare by aphids is reduced by earthworms

Soil organisms affect plant growth and chemistry and have subsequent effects on aboveground herbivore performance. However, whether herbivores discriminate between plants exposed to different soil organisms when colonizing their host plants is largely unexplored. In a greenhouse study, Tanacetum vulgare L. (Asteraceae) growing in a ruderal plant community in the presence and absence of arbuscular mycorrhizal fungi (AMF) and earthworms [Aporrectodea spp. (Haplotaxida: Lumbricidae)] was colonized by aphids [Myzus persicae Sulzer (Hemiptera: Aphididae)]. The aphids preferred to colonize plants without earthworms in the soil, and the numbers of aphids remained lower on the plants with earthworms, irrespective of the presence of AMF. Although the N, C, and P concentrations of the shoots were not affected by the soil organisms, AMF increased total aboveground biomass, total N, C, and P content, and photosynthetic activity (measured as electron transport rate) in the leaves under high light intensity. These results suggest that earthworms affect chemical cues that are used by aphids to judge host quality prior to feeding. Discrimination between plants with and without exposure to earthworms by aboveground herbivores is a novel aspect of plant‐mediated interactions between below‐ and aboveground organisms.     

Composition of the essential oils of Tanacetum argyrophyllum (C. Koch) Tvzel. var. argyrophyllum and Tanacetum parthenium (L.) Schultz Bip. (Asteraceae) from Turkey

The aerial parts of Tanacetum argyrophyllum (C. Koch) Tvzel. var. argyrophyllum and T. parthenium (L.) Schultz Bip. were hydro-distilled to produce the oils in the yields of 0.78% (v/w) and 0.43% (v/w), respectively. The oils were analysed by GC and GC/MS. Twenty-two and twenty-three components were identified representing 94.2% and 90.1% of the oils, respectively. The main compounds of T. argyrophyllum were cis-thujone (69.9%), trans-thujone (5.6%) and 1,8-cineole (3.2%), whereas camphor (56.9%), camphene (12.7%) and p-cymene (5.2%) were the major constituents of T. parthenium.     

Identification of antioxidant phenolic compounds in feverfew (Tanacetum parthenium) by HPLC-ESI-MS/MS and NMR

Antioxidant polyphenolic acids in the medicinal herb feverfew (Tanacetum parthenium) were isolated through in vitro bioassay-orientated antioxidant tests in response to 1,1-diphenyl-2-picrylhydrazyl (DPPH*) free radical scavenging and Fe(2+)-chelating activities. Purification of the active compounds and their structural elucidation involved a variety of techniques including open-column chromatography, HPLC, GC-MS, LC-MS and NMR. Major compounds with potent DPPH* scavenging activities were characterised as 3,5-, 4,5- and 3,4-di-O-caffeoylquinic acids (DCQAs). This is the first report of DCQAs found in feverfew.     

First total synthesis of structurally unique flavonoids and their strong anti-inflammatory effect

The first total synthesis of structurally unique flavonoids 1a and 1b is described. These compounds showed very strong anti-inflammatory effect against delayed hypersensitivity in a mouse model.     

Acaricidal properties of Artemisia absinthium and Tanacetum vulgare (Asteraceae) essential oils obtained by three methods of extraction

Essential oils of Artemisia absinthium L. and Tanacetum vulgare L. were extracted by three methods, a microwave assisted process (MAP), distillation in water (DW) and direct steam distillation (DSD), and tested for their relative toxicity as contact acaricides to the two spotted spider mite, Tetranychus urticae Koch. All three extracts of A. absinthium and of T. vulgare were lethal to the spider mite but to variable degrees. The LC50 obtained from the DSD oil of A. absinthium was significantly lower (0.04 mg/cm2) than that of the MAP (0.13 mg/cm2) and DW (0.13 mg/cm2) oil of this plant species. DSD and DW extracts of T. vulgare were more toxic (75.6 and 60.4% mite mortality, respectively, at 4% concentration) to the spider mite than the MAP extract (16.7% mite mortality at 4% concentration). Chromatographic analysis indicated differences in composition between the more toxic DSD oil of A. absinthium and the other two extracts of this plant, indicating that a sesquiterpene (C15H24) compound present in the DSD oil and absent in the other two may enhance the toxicity of the DSD oil. Chemical analysis of the T. vulgare extracts indicated that beta-thujone is by far the major compound of the oil (>87.6%) and probably contributes significantly to the acaricidal activity of the oil.     

Electron dense substance of thylakoids in chloroplasts of tansy Tanacetum vulgare L

An intrathylakoid electron opaque substance, further referred to as loculin, is found in 80-90 % of thylakoids of tansy leaf mesophyll chloroplasts at the stage of flower bud formation and flowering. Upon conventional isolation of chloroplasts in aqueous solution, and fixation in osmium solution alone, loculin is not retained in thylakoids. Preliminary fixation of leaves in glutaraldehyde makes it possible to isolate chloroplasts without injuring the envelope and stroma (glutar chloroplasts), and loculin is retained in thylakoids under these conditions. Upon prolonged incubation of glutar chloroplasts (for 24 h), loculin leaves thylakoids in the form of drops concentrating on the chloroplast envelope. Upon crossing the thylakoid membrane and chloroplast, loculin properties remain unchanged. It is assumed that loculin is an important metabolite necessary for active growth.     

Structure of tanacetan,a pectic polysaccharide from tansy Tanacetum vulgare L

Tanacetan TVF was found to have a branched structure with a backbone of linear -1,4-D-galacturonan. The ramified regions consist of linear -1,2-L-rhamno--1,4-D-galacturonan as the core. The side chains appear to attach to the 4-position of the L-rhamnopyranose residues. They are present as single -galactopyranose residues or a branching -1,4-galactopyranan bearing 4,6-substituted -D-galactopyranose residues as branched points. In addition, the ramified regions contain side chains of a branched -1,5-arabinofuranan possessing 2,5- and 3,5-substituted -L-arabinofuranose residues as branching points. Some side chains of rhamnogalacturonan appear to be arabinogalactan which contains branched sugar chains of -1,5-arabinofuranan attached to the linear chains of -1,4-galactopyranan by 1,3- and 1,6-linkages. The residues of -L-arabinofuranose seem to occupy the terminal positions of the arabinogalactan side chains.     

Genetic and morphological diversity of Finnish tansy (Tanacetum vulgare L., Asteraceae)

 Morphological and genetical differences of twenty Finnish tansy (Tanacetum vulgare) genotypes were studied. The genotypes were distinguishable by morphology, and number of flower heads, which correlated positively with height of the plant and height of the corymb. The mean nuclear DNA content in the leaves of tissue-cultured tansy plantlets was 8.86 pg, and variation between genotypes was 27%. The genotypes were also distinguishable on the basis of their RAPD patterns. The RAPD and morphological data were subjected to analysis of principal components, according to which the genotypes were separated into two main groups. Group I included 7 genotypes from southern Finland and 2 genotypes from the eastern lake district. They started to flower later, had more flower heads and nodes per stem, and the corymb was longer than in the other genotypes that originated in the western and central part of Finland and formed group II. Our data are supported by previous studies and suggest that group I may represent native tansy populations in Finland, whereas group II may represent tansy genotypes that have been transported to other parts of Finland through agriculture and attempts of domestication. Received: 19 November 1997 / Accepted: 25 November 1997