Band 58, Heft 7-8

Thirty-six alkaloids were identified in the organs of Datura ceratocaula by GC/MS. Thirtythree of them have not been previously reported for the species. Furthermore, a new tropane ester was tentatively identified as 3-(3′-formyloxytropoyloxy)tropane on basis of its mass spectral fragmentation. Hyoscyamine was the main alkaloid in the plant organs.

The composition of essential oils of Leutea glaucopruinosa (Rech.f.) Akhani & Salimian comb, nov., and Zeravschania (Boiss. & Hausskn.) Salimian & Akhani comb. nov. were analysed by GC-MS. 49 compounds are identified in the former and 33 compounds in the latter, comprising a total of 76 compounds in both species. Both species were originally described under Peucedanum, which are transferred in this paper into Leutea and Zeravschania, respectively. The chemical compounds of the essential oils show that there are only seven common compounds between two species. The major compounds of L. glaucopruinosa are mostly monoterpene hydrocarbons and oxygenated monoterpenes, in which α-pinene (31.5%), sabinene (9.7%), β-pinene (9.2%), exo-fenchyl acetate (4.5%) are dominant. In Z. pastinacifolia sesquiterpene hydrocarbons and phenylpropanoids dominate with β-bisabolene (37.3%), 3,1-butyl-1.2-dimethoxy benzene (14.9%), 10,11-dimethylbicyclo[6.3.0]undec-(8)-en-9-one (12.9%), 4-t-butyl-1,2-dimethoxy benzene (6.8%), (E)-asarone (5.1%) and elemicine (4.1%) as major compounds.

Cuticular waxes of Viscum album subspecies and of V. cruciatum have been examined for their micromorphology and chemical composition. Wax crystalloids occur preferably as irregular platelets and rodlets, while deviant structures are found in small areas. Among the triterpenoids forming the wax layer, oleanolic acid is prevailing with some 80%. The quantitative composition of the long-chain aliphatics, which comprise several classes, is rather variable. Flavonoid aglycones, occurring as very minor components of the cuticular waxes, comprise the flavonols kaempferol and quercetin and a series of their methyl derivatives, in some taxa also the flavanone naringenin. Neither the crystalloid structures nor the chemical composition of the wax allow to discriminate the 2 species, or male and female plants, or plants grown on conifers or on dicotyledoneous hosts.

A new phenylethanoid tetraglycoside, physocalycoside (2), was isolated from the aerial parts of Phlomis physocalyx. Its structure was identified as 3-hydroxy-4-methoxy-β-phenyl-ethoxy-O-[α-ʟ-rhamnopyranosyl-(1→2)-α-ʟ-rhamnopyranosyl-(1→3)]-4-O-feruloyl-[β-ᴅ- glucopyranosyl-(1→6)]-|β-ᴅ-glucopyranoside, on the basis of spectroscopic evidence. In addition, one known iridoid glucoside, lamiide (1) and five known phenylethanoid glycosides, wiedemannioside C (3), verbascoside (= acteoside) (4). leucosceptoside A (5), martynoside (6), and forsythoside B (7) were also characterized. Compounds 2-7 demonstrated radical scavenging properties towards the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical.

Salsolidine and 2-methyl,1,2,3,4-tetrahydro-β-carboline were isolated from Haloxylon articulatum (growing in Egypt) besides the known carnegine and N-methylisosalsoline. 2- methyl,1,2,3,4-tetrahydro-β-carboline was recorded for the first time in the genus Haloxylon. The alkaloids were identified by spectroscopic methods (MS, 1D and 2D NMR).

A new highly oxygenated iridoid glucoside, urphoside B (1) was isolated from the Veronica pectinata var. glandulosa together with seven known iridoid glucosides, aucubin, catalpol, veronicoside, catalposide, verproside, amphicoside and 6-O-veratroyl catalpol. The planar as well as the stereo structures of the isolated compounds were determined by means of extensive 1D- and 2D-NMR spectroscopy and confirmed by HR-Mass.

Four new triterpenoid saponins, 3-O-{[β-ᴅ-glucopyranosyl-(1→2)]-β-ᴅ-galactopyranosyl}- 11α-methoxy-23-hydroxylongispinogenin, 3-O-{[β-ᴅ-glucopyranosyl-(1→2)]-β-ᴅ-galactopyranosyl}- 11α-methoxy-23,29-dihydroxylongispinogenin, 3-O-{[β-ᴅ-glucopyranosyl-(1→2)]-β-ᴅ-galactopyranosyl}- 29-hydroxysaikogenin F and 3-O-{[β-ᴅ-glucopyranosyl-(1→2)]-β-ᴅ-galactopyranosyl}- saikogenin F, have been isolated from Atriplex semibaccata. The structures were determined primarily by NMR spectroscopy. The assignment of NMR signals was performed by means of 1 H- 1 H COSY, ROESY, HMQC and HMBC experiments.

Polygala paniculata L. yielded the xanthones 1-hydroxy-5-methoxy-2,3-methylenedioxyxanthone (1) and 1,5-dihydroxy-2,3-dimethoxyxanthone (2), together with coumarin murragatin and flavonol rutin. Their structures were established by chemical and spectroscopic methods (EIMS, IR, 1 H and 13 C NMR, NOE difference spectroscopy). By posterior analysis of an apolar crude extract using high resolution gas chromatography coupled to mass spectrometry (HRGC-MS) it was possible to characterize two sterol (spinasterol and Δ 25 -spinasterol) and the minor 1-hydroxy-2,3,5-trimethoxyxanthone (3). Thus, the xanthone 3 was confirmed through of co-injection HRGC-MS of the respective extract with a certified standard obtained by methylation of 2 with diazomethane.

The Paraguayan crude drug “tayï pytá” is used to treat cancer, wounds and inflammation. It consist of the bark and trunkwood of Tabebuia heptaphylla (Bignoniaceae). A phytochemical study of the crude drug gave, in addition to previously described naphthoquinones and the known lignans cycloolivil and secoisolariciresinol, three new lapachenol (lapachonone)-, two naphthofuran-, a chromone and a naphthalene derivative. The structures were elucidated by means of high field NMR spectroscopy. The biological activity of the main compound lapachol and the related α-lapachone as well as the lignans cycloolivil and secoisolariciresinol can explain, at least in part, the effect atributed to the crude drug in Paraguayan folk medicine.

The chemical composition of fatty acid methyl esters (FAMEs) from seeds of S. thymbra and S. cuneifolia were analyzed by GC/MS. 7 FAMEs were identified from the seeds of S. thymbra mainly as 9-octadecenoic acid methyl ester (43.9%), hexadecanoic acid methyl ester (11.4%), 9,12,15-octadecatrienoic acid methyl ester (Z,Z,Z) (30.2%), and octadecanoic acid methyl ester (14.1%), while from the seed of S. cuneifolia 10 FAMEs were obtained with the main components, similar to S. thymbra. These were identified as 9-octadecenoic acid methyl ester (10.1%), hexadecanoic acid methyl ester (methyl palmitate, 34.6%), 9,12,15-octadecatrienoic acid methyl ester (Z,Z,Z) (6.3%) and octadecanoic acid methyl ester (1.8%).

The immunomodulatory bioassay-guided fractionation of the oleogum resin of frankincense (Boswellia carterii Bird wood) resulted in the isolation and identification of 9 compounds; palmitic acid and eight triterpenoids belonging to lupane, ursane, oleanane, and tirucallane skeleta were isolated form the resin. These triterpenoids are lupeol, β-boswellic acid, 11-keto-β-boswellic acid, acetyl β-boswellic acid, acetyl 11-keto-β-boswellic acid, acetyl- α-boswellic acid, 3-oxo-tirucallic acid, and 3-hydroxy-tirucallic acid. The structures of the isolated compounds were deduced based on spectroscopic evidences. The lymphocyte transformation assay of the isolated compounds proved that the total extract retained more activity than that of any of the purified compounds.

A new prenylated naphthoquinone dimer named microphyllaquinone (1), a mixture of 6-methoxy- and 7-methoxy-naphtho[2,3-b]-furan-4,9-quinones (2a/2b) and tecomaquinone I (3), were isolated from roots of Lippia microphylla. The structures were elucidated by spectroscopic methods, including detailed 1D and 2D (COSY, NOESY, HMQC, HMBC) NMR data. Unpublished 13 C NMR data of 2a and 2b are reported. The in vitro cytotoxic activity of the isolated compounds was tested against five types of tumor cells.

Fractionation of the chloroform extract from the aerial part of Argemone mexicana led to the isolation of two benzophenanthridine-type alkaloids, N-demethyloxysanguinarine and pancorine; three benzylisoquinoline-type alkaloids, (+)-1,2,3,4-tetrahydro-1-(2-hydroxymethyl- 3,4-dimethoxyphenylmethyl)-6,7-methylenedioxyisoquinoline, (+)-higenamine and (+)-reticuline. Among them, N-demethyloxysanguinarine is a new compound, and (+)-1,2,3,4- tetrahydro-1-(2-hydroxymethyl-3,4-dimethoxyphenylmethyl)-6,7-methylenedioxy-isoquinoline was isolated form a natural source for the first time, to which was assigned a trivial name, (+)-argenaxine. In addition, six known non-alkaloidal compounds were also isolated and identified. All compounds were characterized on the basis of their spectral data and chemical evidences. Some isolated alkaloids from this species were evaluated for their cytotoxicity to human nasopharyngeal carcinoma (HONE-1) and human gastric cancer (NUGC) cell lines. Chelerythrine was found to exhibit significant activity against NUGC cell line, while angoline inhibited both types. (+)-Argenaxine showed moderate activity against the NUGC cell line.

Two new bicyclic acylphloroglucinol derivatives, hypercalyxone A (1-[5,7-dihydroxy-2-methyl- 3-(3-methyl-but-2-enyl)-2-(4-methyl-pent-3-enyl)-chroman-8-yl]-2-methyl-propan-1-one, 1) and B (1-[5,7-dihydroxy-2-methyl-3-(3-methyl-but-2-enyl)-2-(4-methyl-pent-3-enyl)-chroman- 8-yl]-2-methyl-butan-1-one, 2), have been isolated from the petroleum ether extract of the aerial parts of Hypericum amblycalyx, together with two further compounds (1-[5,7- dihydroxy-2-methyl-2-(4-methyl-pent-3-enyl)-chroman-8-yl]-2-methyl-propan-1-one, 3 and 1-[5,7-dihydroxy-2-methyl-2-(4-methyl-pent-3-enyl)-chroman-8-yl]-2-methyl-butan-1-one, 4), which have been described only as semi-synthetic products. In addition, the known triterpene lup-20(29)-en-3-one was obtained. Structure elucidation was based on 1D and 2D NMR studies, as well as on data derived from mass spectrometry. The four acylphloroglucinol derivatives were evaluated for their cytotoxic and antibacterial activity. All compounds showed moderate cytotoxic activity against KB and Jurkat T cancer cells. Especially compounds 3 and 4 exhibited a strong antibacterial activity against different Gram-positive strains.

In addition to the known mutisicoumarin A, the aerial parts of the shrub Mutisia friesiana afforded five new methylphenones, two new 5-methylcoumarins and a new related chromone. Their structures were elucidated by spectroscopic methods. 13 C NMR data for mutisicoumarin A are reported for the first time. Mutisiphenones A and B and mutisicoumarin A showed antifungal activity against the phytopathogenic fungus Cladosporium cucumerinum.

Two new antibiotic metabolites, dacrymenone (1) and VM 3298-2 (2), were isolated from fermentations of a Dacrymyces sp., and their structures were determined by spectroscopic techniques. Dacrymenone (1) is a new eremophilane sesquiterpenoid while VM 3298-2 (2) is a bis-enol ether between methyl pyruvate and 4,6-dihydroxycyclohexan-1,3-dione. Dacrymenone (1) shows weak antibacterial and antifungal activity while VM 3298-2 (2) exhibits cytotoxic and antifungal activities.

In a search for new fungal compounds inducing apoptosis of the colon cancer derived cell line COLO-320, hormonemate (1) was purified from fermentations of an endophytic fungus isolated from living needles of a Pinus species. The producing strain was identified as Hormonema dematioides by microscopy and ITS rDNA sequence analysis. The structure of hormonemate was determined by spectroscopic methods. The compound exhibited cytotoxic effects against the human colon tumor cell lines COLO-320, DLD-1 and HT-29 as well as five other human cell lines (HL-60, JURKAT, HEP-G2, MCF-7, HeLa S3). It also induced apoptosis in COLO-320 cells as detected by a caspase-activity assay and morphological changes, and it triggered morphological and physiological differentiation of HL-60 cells into granulocytes, which subsequently died by apoptosis.

Sub-aerial parts of Chaerophyllum aureum L. yielded two polyacetylenes, falcarinol (1), falcarindiol (2), three lignans, namely nemerosin (3), deoxypodorhizone (4), deoxypodo-phyllotoxin (5), two phenylpropanoids, 1′-hydroxymyristicin (6) and its angeloyl ester (7). Compounds 6 and 7 were isolated for the first time from plant material and their structures were elucidated by means of extensive 1- and 2-dimensional NMR spectroscopy and high resolution mass spectrometry. In bioautographic tests on TLC plates the dichloromethane extract showed a significant antimicrobial activity. Falcarindiol was identified as the main active principle whereas the phenylpropanoids and lignans showed no activity.

The ability of synthetic protein fragments to survive the degradative action of aminopeptidases and serum proteolytic enzymes can be remarkably enhanced by slight modifications at their N-terminal alpha-amino group. This can be achieved by addition of beta-alanine or amino acids of the d-configuration, amino acids which are seldom found in a living organism. These modifications do scarcely modify the chemical and physical properties of the peptides, and should be preferrred, especially for in vivo tests, to drastic alterations of peptides as produced by dinitrophenylation or dansylation of the amino groups.

Two strains were selected - the lactoso-negative yeast Rhodotorula rubra GED2 and the homofermentative Lactobacillus casei subsp. casei Ha1 for co-cultivation in cheese whey ultrafiltrate (WU) and active synthesis of carotenoids. Under conditions of intensive aeration (1.0 l/1 min, 220 rpm), a temperature of 30 °C, WU with 55.0 g lactose/l, initial pH = 5.5, the carotenoid content in the cells reached a maximum, when the growth of the cultures had come to an end, i.e. in the stationary phase of the yeast. The maxima for dry cell accumulation (27.0 g/l) and carotenoid formation (12.1 mg/l culture medium) did not coincide on the 5 th and 6 th day, respectively. A peculiarity of the carotenoid-synthesizing Rh. rubra GED2 strain, co-cultivated with L. casei Ha1, was the production of carotenoids with high β-carotene content (46.6% of total carotenoids) and 10.7% and 36.9% for torulene and torularhodin, respectively.

The intertidal sponge Hymeniacidon heliophila which survives under intense sunlight contains the antioxidant amino acid l-5-hydroxytryptophan (ʟ-5-HTP) as major constituent. The content of ʟ-5-HTP was determined as (0.45 ± 0.23)% of the dry weight by quantitative NMR-spectroscopical analysis with an internal standard. Other known antioxidants such as flavonoids, carotenoids or tocopherol derivatives were absent. Both the oxidation potential and the concentration of ʟ-5-HTP in H. heliophila correspond to the values observed for flavonoids being major antioxidants in plants. It was shown that ʟ-5-HTP suppresses UVinduced apoptosis in human monocytes at the same concentrations as it occurs in the sponge tissue.

Plagiodera versicolora, a willow beetle living on S. sachalinensis, is found on S. integra during early June in Hokkaido Island, Japan. This insect selects several species of willows (Salix), including S. integra as host plant in Honshu Island of Japan. To determine the reasons for the limited distribution of this beetle on the willows of Hokkaido, the feeding preference of the insect to leaves of S. integra and its constituents was performed. Feeding-bioassay guided fractionation of an 80% aqueous acetone extract of fresh leaves of Salix integra to Plagiodera versicolora resulted in isolation of feeding stimulant and antifeeding constituents. Chlorogenic acid (1) and 3,5-dicaffeoyl quinic acid (2) were identified as antifeedants and 1,2-di[(9Z,12Z,15Z)-octadeca-9,12,15-trienoyl]-3-β-D-galactopyranosyl-sn-glycerol (MGDG, 3) as feeding stimulants. The feeding test was performed by an agar disk method. The treated agar disks contained sucrose and test sample in different doses. The antifeeding activities of 1 and 2 and stimulant activity of 3 may be one of the reasons for the limited presence of P. versicolora on S. integra in Hokkaido.

Propolis, the resinous product collected by honey bees from plants, is used as folk medicine since ancient time. Recently, immunoregulatory and anti-inflammatory properties of propolis have been published. The detailed mechanisms of actions of propolis and its components on immune cells, however, are still unknown. Therefore, we studied the effects of different propolis extracts, of the flavonoids hesperidin and quercetin as well as of caffeic acid phenethyl ester (CAPE) on basic human immune cell functions. In detail, we measured the effects on DNA synthesis and production of different types of cytokines, namely IL-1β, IL-12, IL-2, IL-4, IL-10 and TGF-β1, of mitogen-activated peripheral blood mononuclear cells (PBMC) as well as of purified T lymphocytes. Our data clearly show that propolis as well as its constituents studied are capable of dosedependently suppressing phythemagglutinin (PHA)-induced DNA synthesis of PBMC and T cells. Moreover, cytokines produced by monocytes/macrophages (IL-1β, IL-12), by Th1 type (IL-2) as well as Th2 type (IL-4) lymphocytes were found to be also suppressed, whereas the production of TGF-β1 by T regulatory cells was ascertained to be increased. These data convincingly demonstrate that propolis has a direct regulatory effect on basic functional properties of immune cells which may be mediated by the Erk2 MAP-kinase signal pathway. Thus, the bee product propolis can be considered as a powerful natural anti-inflammatory medicine influencing different types of immune-responses probably via immunoregulatory T cells.

The quinic acid derivatives (including 4-feruoyl quinic and 5-ferruoyl quinic acids characterized for first time in propolis samples) and other phenolic compounds were quantified in thirteen Brazilian propolis samples by HPLC analysis. For chemometrical analysis, the distribution of quinic acid derivatives and other phenolic compounds were considered. The results suggest that the Brazilian propolis with floral origin from Citrus sp. have the highest concentration of the quinic acid derivatives (between 11.0 to 58.4 mg/mg of the dried crude hydroalcoholic extract) and therefore would probably show a more effective hepatoprotective activity.

A membrane-bound sialidase was isolated from blood stream (BS) Trypanosoma evansi partially purified and characterized. The enzyme is a glycosyl phosphatidyl inositol (GPI) membrane anchored protein. It was solubilized from T.evansi cells recovered from infected camel blood by detergent treatment with Triton CF 54 and partially purified by a series of chromatography steps. The enzyme was optimally active at pH 5.5 and 37 °C. It had a Kᴍ and V max values of 4.8 x 10 -6 ᴍ and 3.75 x 10 -6 mol/min.mg protein with Neu5Acα2, 3lac as substrate respectively. The Kᴍ and V max values with fetuin (4-nitrophenyl-oxamic acid) as substrate were 2.9 x 10 -2 ᴍ and 4.2 \ 10 -3 mol/min.mg protein in the same respect. Kinetic analysis with methly umbelliferyl sialate (MU-Neu5Ac) gave Kᴍ and V max values of 0.17 mᴍ and 0.84 mmol/min.mg protein respectively. The T. evansi SD could hydrolyse internally linked sialic acid residues of the ganglioside GM 2 , but was inactive towards colomic acid, and Neu5Ac2, 6. lac. When ghost red blood cell (RBC) was used as substrate, it desialylated the RBC in the following order of efficiency; mouse, rat, camel, goat, and dog. Similarly, cerebral cells isolated from BalbC mouse was desialylated by the T. evansi SD. Inhibition studies using 2-deoxy-2, 3 didehydro-N-acetyl neuraminic acid (NeuAc2, 3en) against MU-Neu5Ac revealed a competitive inhibition pattern with K i of 5.8 μm. The enzyme was also inhibited non-competitively by parahydroxy oxamic acid (pHOA), and competitively by N-ethylmaleimide and N-bromosuccinate with K i values of 25, 42, and 53 μm, respectively. It was activated by Mg 2+ ion and inhibited by Cu 2+ and Zn 2+ .

Astragalus membranaceus (Fisch.) Bunge has been was successfully acclimated in Central Europe. We report the content of isoflavones and some other polyphenolic compounds in roots and aerial parts that have been analyzed by means of TLC and HPLC. The total amount of isoflavones in leaves, was 0.55 mg g -1 dry weight, and of the flavonols - up to 3.54 mg g -1 . In the roots isoflavonoid content was extremely variable, but reached 3.04 mg g -1 , whereas flavonols content was 0.49 mg g -1 .

In cell suspension cultures of Taxus chinensis, 40 mg/l fungal elicitor from Aspergillus niger and 20 μm HgCl 2 elicited 5.7 and 3.6 mg/l taxol, which was a 9-fold and 5- fold increase vs. compared with the control, respectively. The fungal elicitor induced hydrogen peroxide (H 2 O 2 ) accumulation but HgCl 2 did not, indicating that H 2 O 2 was not necessary for enhancement of taxol induced by elicitor. Compared with the treatment with fungal elicitor alone, exogenous catalase, ascorbic acid, diphenylene iodonium and superoxide dismutase induced a 0.45, 0.4, 0.7 and 1.4-fold H 2 O 2 , but elicited taxol production, which was 0.98, 1.2, 1.1 and 0.9-fold, respectively, vs. non-treated cells. Elicitor-induced taxol production was not accorded with the amount of H 2 O 2 production.