Extracts of Cynomorium songaricum protect human neuroblastoma cells from beta-amyloid25-35 and superoxide anion induced injury.

In Traditional Chinese Medicine a number of herbs are used to alleviate age-related diseases including memory impairment and dementia, among them stems of Cynomorium songaricum, Cynomoriaceae. In this study, we evaluated the protective effect of different extracts of aerial parts of C. songaricum on amyloid-beta peptide (Abeta) and hypoxanthine/xanthine oxidase induced cell death in SK-N-SH neuroblastoma cells. Abeta (20 microM) as well as superoxide anions generated by the hypoxanthine/xanthine oxidase system both reduced cell viability to about 60%. The methanolic extract of C. songaricum attenuated Abeta induced cell death at concentrations of 100 and 10 microg/ml, an even stronger effect was observed for the ethyl acetate fraction obtained from the crude methanolic extract. On the other hand, the dichloromethane as well as water fractions showed no protective effects. In order to further analyze the protective mode of action, the ability of extracts to protect against superoxide anions induced cell death was also evaluated. In this system, cell viability could again be restored by methanol and ethyl acetate extracts, the latter showingsignificant protective effects even at concentrations as low as 0.1 microg/ml.

A cell-based high-throughput assay system reveals modulation of oxidative and nonoxidative glucose metabolism due to commonly used organic solvents.

A 96-well format screening system was generated to quantify changes in nonoxidative glucose metabolism and oxidative pyruvate metabolism. D-Glucose uptake from the supernatant media was quantified by the glucose oxidase method, and L-lactate production of cells was quantified by the lactate dehydrogenase method applied on supernatant media. Mitochondrial membrane potential was quantified using tetramethylrhodamine methyl ester (TMRM) fluorescence, and reactive oxygen species (ROS) formation was determined by quantification of dihydrodichlorofluorescein fluorescence. Adenosine triphosphate (ATP) content of myocytes was determined using the luciferin reaction, and cellular respiration was quantified using commercially available, precoated microtiter plates. These six assays were used to determine the putative influence of organic solvents, namely dimethyl sulfoxide (DMSO), ethanol, methanol, and N-methylpyrrolidone (NMP) at concentrations of 0.01, 0.1, 1.0, and 5.0% (vol/vol), respectively, on glucose and pyruvate metabolism after 4 and 24 hours. In summary, all solvents induced significant changes in regard to one or several of the parameters evaluated, affecting cellular glucose uptake, glycolysis, mitochondrial metabolism, or oxidative phosphorylation. Accordingly, this comprehensive HTS evaluation should enable researchers to choose specific organic solvents on a rational basis to avoid nonspecific effects in cultured cells and tissue culture based experimental setups.

Phenolic glycosides from Exostema mexicanum leaves.

Phytochemical investigation of the leaves of Exostema mexicanum led to the isolation of two novel acylated flavonol glycosides 6, 7 and three glycosides 1-4 structurally belonging to the group of 4-phenylcoumarins. One of them, 5-O-beta-D-glucopyranosyl-4'-hydroxy-7-methoxy-4-phenylcoumarin (2), turned out to be new. Furthermore, the 4-phenylcoumarin aglycone 3'-hydroxy-4',5,7-trimethoxy-4-phenylcoumarin (5) was obtained. The in vitro cytotoxicity of 3-5 against the cell line ECV-304 was evaluated; the aglycone 5 was highly cytotoxic, whereas the glycosidic compounds 3 and 4 were inactive.

New prenylated benzoic acid derivatives of Piper hispidum.

Three new 4-hydroxy-benzoic acid derivatives, 4-methoxy-3,5-bis-(3-hydroxy-3-methyl-1-butenyl)benzoate, 3-hydroxy-2-(1-hydroxy-1-methylethyl)-2,3-dihydrobenzofuran-5-carboxylic acid, and 3-hydroxy-2-(1-hydroxy-1-methylethyl)-2,3-dihydrobenzofuran-5-carboxylic acid methyl ester together with eight known compounds, have been isolated from the stems of Piper hispidum. Their structures were elucidated by a detailed spectroscopic analysis. In addition, the cytotoxicity of seven isolated compounds has been evaluated, revealing a moderate activity for three derivatives of dillapiole.

Cytotoxicity of plants used in traditional medicine in Yemen.

Twenty-five extracts obtained from 14 plant species used in the traditional medicine in Yemen have been screened for cytotoxic activity against human ECV-304 cells. Extracts of Dracaena cinnabari, Eucalyptus camaldulensis, Euclea divinorum, Euphorbia cactus, Pulicaria crispa, and Withania somnifera displayed a remarkable activity.

Merrekentrones A-D, ipomeamarone-like furanosesquiterpenes from Merremia kentrocaulos.

Four new furanosesquiterpenes, merrekentrones A (1), B (2), C (3), and D (4), were isolated from the roots and rootstocks of Merremia kentrocaulos. Their structures were elucidated on the basis of spectroscopic data interpretation. In contrast to ipomeamarone (5), the well-known phytoalexin of Ipomoea batatas, 1-4 seem to be normal plant constituents. Merrekentrone A (1) was also detected in the roots of M. guerrichii and M. aurea.

Antileishmanial activities of aphidicolin and its semisynthetic derivatives.

Aphidicolin and a series of semisynthetic aphidicolan derivatives have been identified in in vitro tests as novel drugs with antiparasitic potential. All compounds have been tested against extracellular promastigotes of Leishmania donovani, L. infantum, L. enriettii, and L. major and against intracellular amastigotes of L. donovani in murine macrophages. The compounds showed antileishmanial activity at concentrations in the microgram range (50% effective concentration [EC(50)] = 0.02 to 1.83 microg/ml). The most active derivative (aphidicolin-17-glycinate hydrochloride) had EC(50)s of 0. 2 microg/ml against extracellular and 0.02 microg/ml against intracellular L. donovani parasites. To validate the pharmacological potential of tested drugs, pharmacological safety was determined by testing all compounds against two neoplastic cell lines (squamous carcinoma [KB] and melanoma [SK-Mel]) and against murine bone marrow-derived macrophages as host cells. With minor exceptions only for macrophages, tested aphidicolans did not show significant cytotoxicity (EC(50) > 25.0 microg/ml). Structure-activity relationships of these aphidicolan derivatives are discussed.

New trichothecenes isolated from Holarrhena floribunda.

Bioassay-guided fractionation of an extract of Holarrhena floribunda stem, has led to the isolation of the new trichothecenes, 8-dihydrotrichothecinol A (1), loukacinol A (2), and loukacinol B (3), and the known compounds, trichothecolone (4), trichothecin (5), trichothecinol A (6), rosenonolactone (7), 6beta-hydroxyrosenonolactone (8), and rosololactone (9). The structures were determined by spectral and chemical methods, and absolute configurations were established by a modified Horeau's method using HPLC. Compounds 1 and 6 exhibited significant cytotoxicity against several human tumor cell lines, whereas compound 8 showed moderate and weak antileishmanial activity toward extracellular and intracellular Leishmania donovani, respectively.

Antimicrobial, antitumor and antileishmania screening of medicinal plants from Guinea-Bissau.

Following an ethnobotanical search carried out in Guinea-Bissau, eighteen extracts derived from sixteen medicinal species were screened for antimicrobial, antitumor and antileishmania activity. Significant antitumor activity was found for Holarrhena floribunda against KB (squamous carcinoma), SK-Mel 28 (melanoma), A 549 (lung carcinoma) and MDA-MB 231 (mamma carcinoma) cell lines, with corresponding IC50 values of 7.9, 9.0, 3.4 and 9.9 micrograms/ml. Khaya senegalensis and Anthostema senegalense exhibited a significant activity against Leishmania donovani with IC50 values of 9.8 and 9.1 micrograms/ml, respectively. Most of the extracts showed week or moderate antibacterial and antifungal activity, with MIC values in the range 0.25-1.0 mg/ml. Active extracts were submitted to bioassay-guided fractionation, and the IC50 and MIC of the active fractions were determined.

Aphidicolin glycinate inhibits human neuroblastoma cell growth in vivo.

Aphidicolin is a fungal derived tetracyclic diterpene antibiotic. It is selectively toxic for neuroblastoma (NB) cells in vitro but has no significant effects on the viability of normal human cells and a variety of other tumor entities. We evaluated the antitumoral effects of the water soluble ester aphidicolin glycinate (AphiG) on established human NB xenografts from UKF-NB-3 cells in athymic (nude) mice. Furthermore, we explored the efficacy of direct intraneoplastic and systemic delivery of AphiG. Systemic administration of AphiG (60 mg/kg intraperitoneally, twice per day on 10 consecutive days) significantly suppressed tumor growth but was not able to induce any cures. In contrast, intratumoral AphiG injections (60 or 40 mg/kg/twice a day for 4 days) induced complete tumor regression. Two weeks after the end of treatment no tumor cells were microscopically detectable. Animals were free of tumor for more than 90 days. Histologic examination of inner organs and bone marrow did not reveal any apparent toxic effects of AphiG. These data strongly indicate that AphiG deserves further evaluation as a specific treatment for neuroblastoma.

Cloning and insertional inactivation of Streptomyces argillaceus genes involved in the earliest steps of biosynthesis of the sugar moieties of the antitumor polyketide mithramycin.

Two genes (mtmD and mtmE) were cloned and sequenced from the mithramycin producer Streptomyces argillaceus. Comparison with proteins in databases and enzymatic assays after expression in Escherichia coli showed that they encode a glucose-1-phosphate:TTP thymidylyl transferase and a TDP-D-glucose 4,6-dehydratase, respectively. The mtmD gene was inactivated by gene replacement, generating a nonproducing mutant that accumulates a tetracyclic compound designated premithramycinone. The identification of premithramycinone reveals new aspects of the mithramycin biosynthetic pathway and suggests that at least some glycosylations occur before breakage of the fourth ring.