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Continued examination of the stem bark of Newbouldia laevis afforded three minor phenylethanoid glycosides, designated as newbouldiosides D – F. Their structures were elucidated by spectroscopic methods as β-(3,4-dihydroxyphenyl)ethyl 5-O-syringoyl-β-D-apiofuranosyloxy-(1 → 2)-O-[α-L-rhamnopyranosyl-(1 → 3)]-6-O-E-sinapoyl-β-D-glucopyranoside, β-(3,4-dihydroxyphenyl)ethyl β-D-apiofuranosyloxy-(1 → 2)-O-[α-L-rhamnopyranosyl-(1 → 3)]-6-O-E-sinapoyl-β-D-glucopyranoside, and β-(3,4-dihydroxyphenyl)ethyl β-D-apiofuranosyloxy-(1 → 2)-O-α-L-rhamnopyranosyl-(1 → 2)-6-O-E-sinapoyl-β-D-glucopyranoside, respectively. These metabolites are the first members possessing a sinapoyl structural element. In addition, the series of naturally occurring flavonoids is extended by the identification of 3′,4′,5,7-tetrahydroxy-5′-methoxyflavanone and apigenin-5-O-α-L-rhamnopyranosyl-7-O-β-D-glucopyranoside obtained from leaf extracts of Markhamia zanzibarica and aromadendrin-7-O-(2″-O-formyl)-β-D-glucopyranoside isolated from Spathodea campanulata. The latter compound is the first example of a flavonoid possessing a formylated glucosyl moiety.

As the sixth-largest Angiosperm family, Lamiaceae contains more than 245 genera and 7886 species that are distributed worldwide. It is also the third-largest family based on the number of taxa in Turkey where it is represented by 46 genera and 782 taxa with a high endemism ratio (44%). Besides, Lamiaceae are rich in plants with economic and medicinal value containing volatile and nonvolatile compounds. Many aromatic plants of Lamiaceae such as Salvia, Sideritis, Stachys, Phlomis, and Teucrium species are used in traditional herbal medicine throughout Turkey as well as in other Mediterranean countries. Salvia (Sage tea “Adaçayı”), Sideritis (Mountain tea “Dağçayı”), Stachys (Hairy Tea “Tüylü Çay”), and Phlomis (Turkish sage “Çalba or Şalba”) are the largest genera that are used as herbal teas. This review focuses on the volatile and nonvolatile compounds of Lamiaceae from the genera Phlomis and Eremostachys of the subfamily Lamioideae with emphasis on iridoids, phenylethanoid glycosides, and essential oils.

Five new compounds, a flavonol glycoside ( 1), a megastigmane ( 2), 2 cyclohexylethanoids ( 3, 4), and a phenylethanoid derivative ( 5), together with 15 known compounds ( 6– 20) including flavonoid glycosides, cyclohexylethanoids, and phenolic compounds, have been isolated from Cornus sanguinea drupes. All the structures have been determined by 1D and 2D NMR spectroscopic analysis and mass spectrometry data. The antioxidant capability of the most representative isolated compounds was evaluated in the hydrogen peroxide (H2O2)-induced premature cellular senescence model of human dermal and gingival fibroblasts. Several derivatives counteracted the increase of reactive oxigen species (ROS) production in both cellular models. Among the most promising, compounds 8, 14, and 20 were able to counteract cell senescence, decreasing the expression of p21 and p53. Furthermore, compound 14 decreased the expression of inflammatory cytokines (IL-6) in both cell models and counteracted the decrease of collagen expression induced by the H2O2 in dermal human fibroblasts. These data highlight the anti-aging properties of several isolated compounds from C. sanguinea drupes, supporting its possible use in the cure of skin or periodontitis lesions.

International audience; Dolichandrone spathacea (L.f.) K. Schum. named “mangrove trumpet tree” is a common smooth tree growing wild in riverbanks and mangroves of the Asia-Pacific area. The leaves of this traditional medicinal plant are used in different countries to treat oral thrush (as mouthwash), flatulence and bronchitis (juice of leaves) [1, 2, 3]. The sole report on the plant found in the literature mentioned the detection of flavonoids, triterpenes and tannins [4].Chemical investigation of the methanolic extract of leaves of a sample collected in Vietnam, furnished 16 iridoids, 3 saponins, 3 phenylethanoid glycosides, 4 flavonoid glycosides, 3 monoterpenic acids, 5 phenolic acids and 1 megastigman glucoside. Their structures were established by spectral means, particularly extensive 2D NMR and MS-MS, and from partial hydrolysis. Among these compounds, five are new.With regard to the traditional use of the leaves against various infections and according to literature results on the antimicrobial activity of the methanolic extract against MRSA [4], we set to study our material along these lines. Thus, the antimicrobial activity of five extracts and of all isolated compounds was evaluated against 22 microorganisms using methods such as bioautography, measurement of MIC (minimum inhibitory concentration) on microplates, and screening using a multiple inoculator (Steers) [5]. At variance with most of the literature, we chose to determine the MIC against a large spectrum of microorganisms in order to discover extracts with a broad-scale activity.The results reveal for the first time the presence in this species of several new natural compounds with an interesting level of antimicrobial activity (the lowest MIC = 31.2 µg/ml). The synergy (activity for mixtures of compounds) and in vivo studies with these compounds may be interesting in the future for a valorization approach.

Verbascoside is a phenylethanoid glycoside widely distributed in nature, especially among the order Lamiales, occurring in numerous plants that are constituents of folk medicine preparations. This natural compound, previously isolated by our group from the ethyl acetate extract of Lantana trifolia using the gradient approach in countercurrent chromatography, was now isolated from the butanol extract of the same plant and from Lippia alba f. intermedia (Verbenaceae) using countercurrent chromatography in either gradient or isocratic elution modes. The ethyl acetate extract of L. alba, rich in phenylethanoids and flavonoids, was fractionated using countercurrent chromatography in the step-gradient elution approach. The four-step solvent system was composed of n-hexane-ethyl acetate-n-butanol-water (4 : 10 : X : 10), where X = 1 (solvent system A), 3 (solvent system B), 5 (solvent system C), and 7 (solvent system D), and allowed for the isolation of verbascoside along with other phenylethanoids and flavonoids from both plants. Verbascoside and 2'-O-β-apiosylverbascoside were further isolated from the n-butanol extract of L. trifolia using the solvent system ethyl acetate-n-butanol-water 10 : 2 : 10 on an isocratic run. The difference in the complexity of the two plant extracts demanded different purification steps, which included a second high-speed countercurrent chromatography purification using the isocratic elution mode.

In traditional medicine, flowers and aerial parts of Lamium album are assigned by their anti-inflammatory, antiseptic, and mucolytic activities, and are used in chronic bronchitis and pharyngitis as well as skin, vaginal, and cervical inflammation.The aim of the present study was to compare effects of ethanolic extracts prepared from flowers and aerial parts of L. album on selected functions of human neutrophils, which are involved in an inflammatory response. In order to identify the compounds engaged in the anti-inflammatory activity of extracts, the bioassay-guided isolation of compounds was performed based on the inhibition of cytokine secretion by stimulated neutrophils.The extracts were phytochemically characterized with the HPLC-DAD-MSn method. The inhibition of reactive oxygen species production by formyl-met-leu-phenylalanine- or phorbol 12-myristate 13-acetate-stimulated neutrophils was determined using luminol- or lucigenin-dependent chemiluminescence. The effect on myeloperoxidase secretion by neutrophils was established spectrophotometrically. The levels of cytokine (interleukin 8, TNF-α) production after extract treatment was measured by ELISA.The most abundant constituents of extracts were phenylpropanoids, iridoids, flavonoids, and phenolic acids. Both extracts at concentrations of 25 and 100 µg/mL significantly inhibited reactive oxygen species production, and myeloperoxidase and interleukin 8 secretion. The phenylethanoid glycosides, such as lamiusides A, B, and C as well as 6″-O-β-D-glucopyranosylmartynoside, were isolated and identified. The cells treated with 6″-O-β-D-glucopyranosylmartynoside and lamiuside B produced 29.47 ± 7.11 % and 64.67 ± 5.25 % of interleukin 8, respectively, compared to non-treated control cells.Our results support the traditional use of L. album and indicate it as a potential source of natural anti-inflammatory constituents, such as phenylpropanoids.

We have investigated the protective effects of echinacoside (ECH), one of the phenylethanoid glycosides, on H(2)O(2)-induced cytotoxicity in the rat pheochromocytoma cell line (PC12 cells). Our data show that application of ECH to H(2)O(2)-injured PC12 cells (HIPCs) increased cell viability and decreased the apoptotic ratio. Flow cytometry (FCM) and laser scanning confocal microscopy (LSCM) analysis suggested that ECH exerted its inhibitory effects on the formation of reactive oxygen species (ROS) and the accumulation of intracellular free Ca(2+) ([Ca(2+)]i). In addition, ECH elevated the mitochondrial membrane potential (MMP) in HIPCs. Furthermore, Western blot analysis revealed that ECH prevented an H(2)O(2)-induced increase of the Bax/Bcl-2 ratio by down-regulating Bax protein expression and upregulating Bcl-2 protein expression. In summary, ECH showed significant neuroprotective effects on HIPCs through the mitochondrial apoptotic pathway, and could be a potential candidate for intervention in neurodegenerative diseases such as Alzheimer's and Parkinson's disease.

In the Flora of Turkey, the genus Digitalis is represented by nine species [1]. In this research aerial parts of Digitalis davisiana Heywood was studied phytochemically on the basis of LXR (Liver x receptor) ligand activity, free radical scavenging and cytotoxic activities guided fractionation. Methanol extract applied to polyamide column chromatography for fractionation. Due to examination of mentioned bioactivity tests, active fractions were detected. Radical scavenging activity was tested against DPPH, NO, SO radicals. IC50 values of methanolic extract for tested radicals were found to be; 133.8, 309.9, 1560 µg/mL, respectively. Cytotoxic activity was tested against HEp-2, HepG2 and 3Y1 cell lines by MTT method [2]. IC50 values were 50.8, 211.4 and 94.7 µg/mL respectively. LXR ligand activity was tested by a LXRα luciferase reporter assay on HEK293 cells [3]. Since methanol extract showed cytotoxicity, LXR ligand activity couldn't be determined. When fractions tested in lower concentrations moderate activity was detected. After evaluating the bioactivity test results, isolation studies started from active, phenylethanoid glycosides rich fractions. Studies resulted in the isolation of six phenylethanoid glycosides. Lugrandoside, maxoside and purpureaside E were identified by extensive NMR techniques. Structural determination studies on the other active compounds are in progress. Acknowledgements: Vahap Murat Kutluay was supported by research grant from The Scientific and Technological Research Council of Turkey (2214/A,2211). References: [1] Davis PH (1978) Flora of Turkey and the East Eagean Islands, University Press, Edinburg, Vol 6. [2] Saracoglu I, et al. Studies on constituents with cytotoxic and cytostatic activity of two Turkish medicinal plants Phlomis armeniaca and Scutellaria salviifolia. Biol Pharm Bull 1995; 18: 1396 – 1400 [3] Kotani H, et al. Identification of a naturally occurring rexinoid, honokiol, that activates the retinoid X receptor. J. Nat. Prod. 2010; 73: 1332 – 1336.

The metabolomic approach and data-mining gains increasing significance in phytochemical analysis. The aim of the current work was to examine the effects of the N source concentration and ammonium-nitrate ratio on the pattern of phenylethanoid glycosides (PG) of the tissue cultures of Plantago lanceolata L. The experiment was carried out using a full factorial experimental design, the metabolome was investigated with LC-ESI-MS3; using modified Murashige Skoog media, 10, 20, 40, 60 mM total N source concentrations and 0, 0.11, 0.20, 0.33 ammonium-nitrate ratios were tested. Phenomena were sought using accepted data-visualization and data-mining methods. The effects of the treatment on the individual metabolites was also examined. From the tissue culture, 16 natural products could be putatively identified as PGs. The concentration of these reacted very differently to the treatments, despite the compounds were of the same metabolite group. Plantamajoside and acteoside were detected as major PGs. Their maximal concentration was 3.54 ± 0.83% and 1.30 ± 0.40%, respectively, on media 10 (0.33) and 40 (0.33). Most of the examined 89 abundant natural products were affected by the composition of the media. The N source concentration and the ammonium-nitrate ratio significantly influenced the concentration of 42 and 10 compounds, respectively. For cumulation of many compounds, compositions, such as 10 (0), 10 (0.11) and 40 (0.33) were found to be optimal, which significantly differ from the widely used Murashige Skoog medium – 60 (0.33). Computer simulation has shown, that using one-factor-at-a-time experimental design instead of full-factorial would have led to suboptimal yields with respect to many natural products. If the N source is optimized first, followed by the ammonium-nitrate ratio, this bias is significantly less during one-factor-at-a-time experimental design.

Anti-plasmodial activity-guided fractionation of Phlomis brunneogaleata (Lamiaceae) led to the isolation of two new metabolites, the iridoid glycoside, brunneogaleatoside and a new pyrrolidinium derivative (2 S,4 R)-2-carboxy-4-( E)- p-coumaroyloxy-1,1-dimethylpyrrolidinium inner salt [(2 S,4 R)-1,1-dimethyl-4-( E)- p-coumaroyloxyproline inner salt]. Moreover, a known iridoid glycoside, ipolamiide, six known phenylethanoid glycosides, verbascoside, isoverbascoside, forsythoside B, echinacoside, glucopyranosyl-(1--G (i)-6)-martynoside and integrifolioside B, two flavone glycosides, luteolin 7- O-beta- D-glucopyranoside ( 10) and chrysoeriol 7- O-beta- D-glucopyranoside ( 11), a lignan glycoside liriodendrin, an acetophenone glycoside 4-hydroxyacetophenone 4- O-(6'- O-beta- D-apiofuranosyl)-beta- D-glucopyranoside and three caffeic acid esters, chlorogenic acid, 3-O-caffeoylquinic acid methyl ester and 5- O-caffeoylshikimic acid were isolated. The structures of the pure compounds were elucidated by means of spectroscopic methods (UV, IR, MS, 1D and 2D NMR, [alpha] (D)) and X-ray crystallography. Compounds 10 and 11 were determined to be the major anti-malarial principles of the crude extract (IC (50) values of 2.4 and 5.9 micrograms/mL, respectively). They also exhibited significant leishmanicidal activity (IC (50) = 1.1 and 4.1 micrograms/mL, respectively). The inhibitory potential of the pure metabolites against plasmodial enoyl-ACP reductase (FabI), which is the key regulator of type II fatty acid synthases (FAS-II) in P. falciparum, was also assessed. Compound 10 showed promising FabI inhibiting effect (IC (50) = 10 micrograms/mL) and appears to be the first anti-malarial natural product targeting FabI of P. falciparum.

Six new compounds, an iridoid glucoside, 6- O-methyl-epiaucubin, three iridoids, artselaenins A, B, C, a phenylethanoid glycoside, artselaeroside A, and a phenylpropanoid glycoside, artselaeroside B, as well as fourteen known compounds, nine iridoid glycosides and five phenylpropanoid glycosides were isolated from the whole plants (including roots, stems, leaves and flowers) of Pedicularis artselaeri. Their structures were identified mainly by spectral evidence.

Ten phenylethanoid glycosides, forsythoside B, acteoside, 2'-acetylacteoside, poliumoside, brandioside, echinacoside, isoacteoside, cistanoside H and E-tubuloside E as well as a new compound, Z-tubuloside E, were isolated from the n-BuOH fraction of Callicarpa dichotoma Raeuschel (Verbenaceae) by bioactivity-guided fractionation using glutamate-injured primary cultures of rat cortical cells as a screening system. These phenylethanoid glycosides significantly attenuated glutamate-induced neurotoxicity at concentrations ranging from 0.1 to 10 microM.

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Scutellaria rupestris subsp. adenotricha (Boiss. & Heldr.) Greuter & Burdet (Lamiaceae) is an herbaceous perennial plant, endemic to Albania and Greece [1]. The methanol extract of the aerial parts afforded eight iridoids, namely catalpol, albidoside, scutellarioside II, dihydrocatalpogenine (C-1) α-epimer/β-epimer, globularin, globularidin, mussaenosidic acid, two known phenylethanoid glycosides, acteoside, martynoside, and one known phenolic derivative, E-p-coumaroylglucoside. The chemical profile of this taxon shows many similarities to that of previously investigated S. albida subsp. albida [2,3], S. albida subsp. colchica [4] and S. goulimii [5], of the S. albida group. References: 1. Bothmer, R. (1985) Nord. J. Bot. 5: 421–439. 2. Gousiadou, Ch. et al. (2007) Phytochemistry, 68: 1799–1804. 3. Gousiadou, C. et al. (2012)J. Enzyme. Inhib. Med. Chem. doi:10.3109/14756366. 4. Calis, I. et al. (1993) Phytochemistry 32: 1213–1217. 5. Gousiadou, C. (2012) BSE. 43:139–141.

The present study describes the isolation and purification of acteoside, 2'-acetylacteoside, poliumoside and brandioside, four phenylethanoid glycosides from Brandisia hancei. We examined their effects on the proliferation of cultured A7r5 rat aortic smooth muscle cells. The proliferative response was measured from the [(3)H]-thymidine incorporation into DNA. All four glycosides suppressed the proliferative response in the presence of 2 % or 5 % fetal bovine serum in a concentration-dependent manner. The rank order of effectiveness for inhibition of cell proliferation was: brandioside > or = poliumoside > 2'-acetylacteoside > or = acteoside. The acetyl group at position 2' of glucose does not seem necessary for the anti-proliferative effects of acteoside and 2'-acetylacteoside, while the hydroxy groups of the aromatic rings appear to play a role. Inhibition of smooth muscle cell proliferation by phenylethanoids indicates that these compounds may have preventative effects on arteriosclerosis.

Radical scavenger activities of 21 phenylethanoid glycosides, including 15 ester derivatives of caffeic, ferulic, vanillic and syringic acid as well as 6 deacyl derivatives were determined by quantifying their effects on the production of reactive oxygen species (ROS) in a luminol-enhanced chemiluminescence assay with formyl-methionyl-leucyl-phenylalanine (FMLP) stimulated human polymorphonuclear neutrophils (PMNs). All phenylethanoids acylated with phenolic acids showed strong antioxidant activity whereas the deacyl derivatives were more than 30-fold less active. Therefore, the antioxidant activity is mainly related to the number of aromatic methoxy and hydroxy groups and the structure of the acyl moiety (C6-C1 or C6-C3). In contrast, modification of the sugar chain or replacement of hydroxy groups by methoxy groups in the acyl or the phenylethanoid moiety is of minor importance. The position of the acyl moiety is without significance. Free caffeic, ferulic, vanillic and syringic acid are less active compared to the phenylethanoid derivatives. This points to the importance of dissociation and lipophilicity of these acids in a cellular test system.

Six new compounds, including five new seco-abietane diterpenoids, 12-deoxy-seco-hinokiol methyl ester (1), 12-deoxy-11,12-dihydro-seco-hinokiol methyl ester (2), callicarpic acid A (3), 9alpha-hydroxycallicarpic acid A (4), and callicarpic acid B (5), and a new phenylethanoid derivative, 4-hydroxyphenethyl tetradecanoate (6), have been isolated from the leaves and twigs of Callicarpa pilosissima, together with 14 known compounds (7-20). The structures of these new compounds were determined through analyses of physical data. 12-Deoxy-11,12-dihydro-seco-hinokiol methyl ester (2), callicarpic acid B (5), and alpha-tocopherol trimer B (15) exhibit antitubercular activities (MICs

The aim of this study was to identify and elucidate the vasorelaxant activity of echinacoside, a phenylethanoid glycoside isolated from the medicinal herb Cistanche tubulosa, and its possible underlying mechanism on isolated rat thoracic aortic rings pre-contracted with phenylephrine (PE, 1 microM) and KCl (60 mM). Echinacoside (30-300 microM) exhibited an acute relaxation in endothelium-intact rings in a concentration-dependent manner, while this relaxation was significantly inhibited in endothelium-denuded condition and in the presence of the endothelial nitric oxide synthase (eNOS) inhibitor, N(W)-nitro-L-arginine methyl ester (L-NNA, 100 microM), an unselective soluble guanylate cyclase blocker, methylene blue (10 microM), the selective sGC inhibitor 1 H-[1, 2, 4]oxadiazolo[4,3- A]quinoxalin-1-one (ODQ, 1 microM); in addition, atropine (1 microM), a selective muscarinic receptor antagonist, partially affected the relaxation. However, the cyclooxygenase inhibitor indomethacin (5 microM) had no influence on the action. Echinacoside enhanced the cyclic guanosine monophosphate (cGMP) production in aortic rings contracted with PE. These results indicate for the first time that echinacoside mediates the endothelium-dependent vasodilator action in rat thoracic aortic rings through nitric oxide (NO)-cGMP pathway.

… Soc Medicinal Plant Res

Plantaginis lanceolatae folium (Plantago lanceolata L., Plantaginaceae) and Plantaginis majoris folium (P. major L.) are well known herbal drugs that have been used for treating inflammation of the mouth and throat. Chemical investigations have detected various constituents. Acteoside as phenylethanoid is a major phenolic constituent and reportedly has antiphlogistic, antihepatotoxic, antioxidant, spasmolytic, antimetastatic and antiviral activity. According to the requirement of Ph Eur Plantaginis lanceolatae folium should contain not less than 1.5% of total ortho-dihydroxycinnamic acid derivatives expressed as acteoside calculated with reference to the dried drug. Leaves of the species P. lanceolata and P. major were collected at the different locations in Croatia and investigated in comparison with Plantaginis lanceolatae folium from commercial origin. The content of acteoside was determined spectrophotometrically according to Ph Eur and ranged from 2.28 to 7.61%. Leaves of P. lanceolata contained a larger quantity of acteoside compared to the leaves of P. major. The results of this investigation indicated that all samples meet the standards of Ph Eur. The quantity of acteoside depended on the various factors, such as plant species investigated, the type of soil, microclimatic conditions and geographic position.

Five new phenylethanoid glycosides, savasides A–E (1–5), along with 6 known ones were isolated from the whole plant of Monochasma savatieri Franch. The structures of 1–5 were elucidated on the basis of spectroscopic data analysis. Moreover, all isolated compounds were evaluated for anticomplement activity through the classical pathway.

The well-known medicinal plant Plantago lanceolata L. (ribwort plantain) was effectively propagated by direct organogenesis from segments of leaves and roots using MS medium supplemented with IAA (11.42 microM), kinetin (9.29 microM) for multiplication and IAA (5.71 microM) for rooting. The plantlets were successfully hardened (80 %) and transferred to field cultivation (100 %). Two lines of callus tissue, derived from leaves and roots, were obtained on MS medium without NH (4)NO (3) and supplemented with 2,4-D (4.52 microM) and kinetin ( 0.46 microM). From plant materials--leaf rosettes from in vitro, leaves from plants in field cultivation obtained by micropropagation, root-derived callus and leaf-derived callus--sixteen phenylethanoid glucosides representing nine different structures were isolated and identified by spectral methods (1D and 2D NMR) as known for the species: lavandulifolioside ( 1), plantamajoside ( 2,) acteoside ( 3); new for the species: leucosceptoside A ( 4), martynoside ( 5), desrhamnosylisoacteoside ( 6), plantainoside D ( 7), desrhamnosylacteoside ( 8) and - 2-(4-hydroxyphenyl)ethyl beta- D-glucopyranosyl-(1-->3)-4- O- trans- and cis- p-coumaroyl-beta- D-glucopyranoside ( 9)--the latter also being found for the first time in nature and named lancetoside. Only plantamajoside ( 2) and acteoside ( 3) were common to all plant materials, the former was the main constituent of calli (1.19 - 2.84 % of dry weight), while the latter was the main constituent of the leaves (1.78 - 10.43 % of dry weight). Flavonoids were present only in plants of field cultivation.

From the aerial parts of Verbascum wiedemannianum Fisch. & Mey. four new phenylethanoid glycosides (wiedemanniosides B - E) were isolated along with four known compounds; 6- O-acetylmartynoside (= wiedemanniosides A), verbascoside, martynoside, echinacoside and leucosceptoside B. The structures of the new compounds were elucidated by employing spectroscopic techniques (NMR, HR-ESIMS, IR, and UV).

Ten compounds, harpagoside (1), 8- p-coumaroylharpagide (2), 8-feruloylharpagide (3), 8-cinnamoylmyoporoside (4), pagoside (5), acteoside (6), isoacteoside (7), 6'- O-acetylacteoside (8), cinnamic acid (9) and caffeic acid (10) were isolated from the storage roots of Harpagophytum procumbens, Pedaliaceae. Compounds 1, 2, 6, 7 and 9 are known for H. procumbens; 3 and 10 were isolated the first time from H. procumbens; compounds 4, 5 and 8 are new natural products. Their structures were elucidated using spectroscopic data (NMR, with NOE, COSY and HMBC experiments, UV, [alpha]). The inhibitory activity of aqueous extracts of the roots of H. procumbens and H. zeyheri as well as the main compounds isolated from H. procumbens was tested on human neutrophile elastase. Although inhibition was comparatively weak a dose-dependence was observed. An IC (50) of 542 microg/mL was determined for the aqueous extract of H. procumbens, but 1012 microg/mL for that of H. zeyheri. 6'- O-Acetylacteoside (8), that is not present in H. zeyheri, inhibited the enzyme with an IC (50) of 47 microg/mL (70 microM), compound 7 with 179 microg/mL (286 microM), 2 with 179 microg/mL (331 microM), 5 with 154 microg/mL (260 microM) and 10, which was also used as reference compound, with an IC (50) of 86 microg/mL (475 microM). The IC (50) values of acteoside, harpagoside, cinnamic acid and stachyose were higher than 300 microg/mL and thus not further determined.

By means of HIV gp41 binding affinity directed chromatographic fractionation, three phenylethanoid glycosides; calceolarioside A ( 1), calceolarioside B ( 2) and acteoside ( 3), along with four hydroxycoumarins; esculin ( 4), fraxin ( 5), fraxetin ( 6) and esculetin ( 7), and one lignan, (-)-olivil ( 8) were isolated from the n-butyl alcohol fraction of Fraxinus sieboldiana var. angustata. Among the isolated compounds, calceolarioside B ( 2) and esculetin ( 7) showed moderate binding affinity on HIV gp41 with IC 50 values of 0.1 mg/ml and 0.5 mg/ml, respectively. Calceolarioside A ( 1), calceolarioside B ( 2), acteoside ( 3), and (-)-olivil ( 8) were isolated for the first time from this plant.

Xanthine oxidase is a key enzyme associated with the incidence of hyperuricemia-related disorders. Repeated chromatography of the enzyme inhibitory part of the water extract of the twigs and leaves of Brandisia hancei (Scrophulariaceae) gave a flavone luteolin, an iridoid glycoside mussaenoside, two beta-sitosterol glycosides daucosterol and beta-sitosterol gentiobioside, and five phenylethanoids arenarioside, brandioside, acteoside, 2'-O-acetylacteoside and isoacteoside. Luteolin and isoacteoside inhibited the xanthine oxidase (XO, EC 1.2.3.2) with the IC50 values at 7.83 and 45.48 microM, respectively. Isoacteoside was found to be the first phenylethanoid that decreased substantially the formation of uric acid by inhibiting competitively xanthine oxidase (Ki value: 10.08 microM). Furthermore, the study suggested that the caffeoylation of the 6'-hydroxyl group of the phenylethanoids was essential for the enzyme inhibitory action.

The five phenylethanoids, acteoside (1), cistanoside F (2), lavandulifolioside (3), plantamajoside (4) and isoacteoside (5) were isolated from the herb of Plantago lanceolata L. (Plantaginaceae). Compounds 1, the major phenylethanoid in the herb of P. lanceolata L., and 4, the major phenylethanoid in the herb of P. asiatica L., showed inhibitory effects on arachidonic acid-induced mouse ear edema.