A new ursane-type triterpene oxoglucopyranoside from Crossopteryx febrifuga.

A new saponin, 3-O-ß-d-3-oxo-glucopyranosyl-ursa-12,20(30)-diene-27,28-dioic acid (1), was isolated from the methanol extract of stem bark of Crossopteryx febrifuga together with the known 3ß-d-glucopyranosyl-ursa-12,20(30)-diene-27,28-dioic acid (2), shanzhiside methyl ester (3), shanzhiside (4), ß-sitosterol (5), ß-sitosterol-3-O-ß-d-glucopyranoside (6), ursa-12,20(30)-diene-27,28-dioic acid (7), hederagenin (8), and oleanolic acid (9). The structures were established by comprehensive interpretation of their spectral data 1D- (1H and 13C), 2D-NMR (1H-1H COSY, HMQC, HMBC), spectroscopic, and electrospray ionisation time-of-flight mass spectrometry analysis. The isolated compounds and extracts were screened for their antibacterial properties. Although the EtOAc and n-BuOH extracts exhibited considerable antibacterial activity against Pseudomonas aeruginosa with minimum inhibitory concentration (MIC) value of 32 µg/mL, compounds 2 and 8 showed moderate activity against Enterococcus faecalis with MIC values of 256 and 128 µg/mL, respectively. The new compound (1) exhibited a moderate antibacterial activity against Staphylococcus aureus with an MIC value of 512 µg/mL.

A dihydrochalcone derivative and further steroidal saponins from Sansevieria trifasciata Prain.

Phytochemical investigation of the aerial parts of Sansevieria trifasciata, one of the most common Dracaenaceae plants, has resulted in the isolation of a new dihydrochalcone derivative named trifasciatine C (1), four previously unreported steroidal saponins as two pairs of inseparable regioisomers: trifasciatosides K/L (2/3), M/N (4/5), together with the known 1,2-(dipalmitoyl)-3-O-ß-D-galactopyranosylglycerol (6), aconitic acid (7), and 1-methyl aconitic acid (8). Their structures were elucidated mainly by extensive spectroscopic analysis (1D and 2D nuclear magnetic resonance) and high-resolution electronspray ionization-mass spectrometry, as well as chemical methods and comparison of their spectral data with those of related compounds. Compounds 2/3 and 4/5 were evaluated for their antiproliferative activity on Hela cells, and no significant effect was observed.

Effects of extracts and compounds from Tricholoma populinum Lange on degranulation and IL-2/IL-8 secretion of immune cells.

Tricholoma populinum Lange is an edible basidiomycete from the family Tricholomataceae. Extracts, fractions, and different metabolites isolated from the fruiting bodies of this mushroom were tested for degranulation-inhibiting activities on RBL-2H3 cells (rat basophils). Dichloromethane extracts decreased degranulation significantly, as did a fraction after column chromatography. In addition, the extract decreased the IL-2 release from Jurkat T cells and the release of IL-8 from HMC-1 human mast cells. The results show the significant effects of extracts of T. populinum on cells of the innate (basophils and mast cells) and adaptive (T cells) immune system and indicate the influence of the mushroom on different immunological processes. As one fraction showed activity, it seems to be possible that it includes an active principle. The compounds responsible for this effect, however, could not be identified as the contents oleic acid (1), ergosterol peroxide (2), and 9,11-dehydroergosterol peroxide (3) showed no effects. Nevertheless, the mushroom could be used for supporting allergy treatment in future studies.

Flavonol Glycosides and Cytotoxic Steroidal Saponins from Furcraea tuberosa (Agavaceae).

Phytochemical analysis of the mature fruits of Furcraea tuberosa (Agavaceae) resulted in the isolation of a new bisdesmosidic spirostanol saponin (1), along with eight known steroidal glycosides (2-9), one known phenolic carboxylic acid ester (10) and three known flavonol glycosides (11-13). The structures of these compounds were assigned using a combination of ID and 2D NMR techniques including ¹H, ¹³C, COSY, TOCSY, HSQC and HMBC NMR, and confirmed by mass spectrometry. Thus the new saponin was elucidated as (25R)-6α-(ß-D-glucopyranosyloxy)-5α-spirostane-3ß-Ο-[(6-Ο-hexadecanoyl)-ß-D- glucopyranoside]. The literature survey revealed that most of the steroidal saponins isolated have shown potent cytotoxic effects against various human cancer cell lines and the results are herein reviewed.

Naghibione; A Novel Sesquiterpenoid with Antiplasmodial Effect from Dorema hyrcanum Koso-Pol. Root, a Plant Used in Traditional Medicine.

Some Dorema species are used in Persian traditional medicine. In the present study the total extract from the roots of Dorema hyrcanum Koso-Pol. was investigated for its in-vitro (pLDH assay) and in-vivo (Peters' 4-days suppressive test) antiplasmodial effects and assessed for cytotoxicity against the normal cell line MDBK (MTT test). The IC50 values for a chloroquine- sensitive (3D7) and a chloroquine- resistant (K1) strain of Plasmodium falciparum were 28.64 and 9.79 µg/mL, respectively. The inhibition percentage of the rodent parasite, Plasmodium berghei, on day 4 in mice was 77.9% and IC50 value on Madin-Darby bovine kidney cells (MDBK cells) was 59.84 µg/mL. The total extract was subjected to a bioassay-guided fractionation protocol based on the in-vivo model which resulted in the isolation of an acetophenon (compound 1), one new sesquiterpenoid; naghibione (compound 2) and two known sesquiterpenoid derivatives (compounds 3, 4). Their structures were elucidated by spectroscopic analysis, including 1D and 2D NMR experiments and ESI-MS. All compounds were evaluated for in-vivo antiplasmodial effect and the results revealed that naghibione showed good suppression activity, inhibiting 68.1 % of the parasite growth.

Cytotoxic saponins from the seeds of Pittosporum angustifolium.

Three new acylated R1-barrigenol triterpene glycosides, 1-3, were isolated from the seeds of Pittosporum angustifolium Lodd. together with four known glycosides, 4-7, containing R1- and A1-barrigenol backbones. On the basis of spectroscopic, spectrometric, and chemical analyses the novel compounds were named pittangretosides N-P and established as 21beta-acetoxy-22alpha-angeloyloxy- (1), 21beta-acetoxy-22alpha-(2-acetoxy-2-methylbutyroyloxy)- (2), and 21beta-(2-methylbutyroyloxy)-22alpha-acetoxy-3beta-[beta-D-glucopyranosyl- (1 --> 2)]-[alpha-L-arabinopyranosyl-(1 --> 3)]-[alpha-L-arabinofuranosyl-(1 --> 4)]-beta-D-glucuronopyranosyloxyolean-12-ene-15alpha, 6alpha, 28-triol (3). Evaluation of the in vitro cytotoxicity against three tumour cell lines and one non-tumourigenic cell line revealed antiproliferative effects with IC50 values in a range of 1.74-34.1 microM.

Cytotoxicity of gypsogenic acid isolated from Gypsophila trichotoma.

BACKGROUND: Gypsophila trichotoma Wend. (Caryophyllaceae) is a medicinal plant which is protected in Bulgaria by the Biodiversity Law. Previous studies have showed the presence of triterpene saponins, sterols, flavonoids, triterpens, etc. OBJECTIVE: Gypsogenic acid, isolated from Gypsophila trichotoma roots, was evaluated for cytotoxic activity. MATERIALS AND METHODS: The structure of the compound was elucidated by spectral methods. The cell survival fraction was determined by the MTT dye reduction assay, performed with some modifications. RESULTS: Gypsogenic acid was tested in a panel of human tumor cell lines and was found to inhibit the proliferation of malignant cells. It was active against leukemic cells with lymphoid (SKW-3 and BV-173) or myeloid phenotype (HL-60, K-562, and LAMA-84), as well as against the EJ bladder carcinoma cell line. Bcr-Abl expressing myeloid cells (LAMA-84 and especially K-562) displayed lower sensitivity. HL-60/Dox cells were less sensitive to gypsogenic acid than the parent cell line, which shows that gypsogenic acid is probably a substrate of MRP-1.

Triterpene glycosides from the leaves of Pittosporum angustifolium.

Phytochemical investigation of the leaves of Pittosporum angustifolium resulted in the isolation and structural elucidation of nine new triterpene saponins, named pittangretosides A-I (1-9), together with a known compound (10). Mainly by NMR and HRESIMS experiments, eight compounds were identified as A1-barrigenol glycosides (1-7, 10), whereas two compounds exhibited an unusual 17,22-seco-backbone of oleanolic acid (8, 9). All compounds were evaluated for their in vitro cytotoxicities against human urinary bladder carcinoma cells (5637). Only compounds with an angeloyl-residue at C-22 of the aglycone (1-4 and 10) showed antiproliferative effects with IC50 values of 4.1, 5.2, 2.1, 17.9, and 2.4 µM, respectively.

Steroidal saponins from the flowers of Dioscorea bulbifera var. sativa.

Eleven steroidal saponins, dioscoreanosides A-K, along with five known congeners, were isolated from the flowers of Dioscorea bulbifera var. sativa. Their structures were established by extensive NMR experiments in conjunction with mass spectrometry. The isolated compounds were tested for cytotoxicity against urinary bladder carcinoma cells (ECV-304 cells). Our results revealed a moderate activity for spiroconazol A (15), pennogenin 3-O-α-l-rhamnopyranosyl-(1→4)-α-l-rhamnopyranosyl-(1→4)-[α-l-rhamnopyranosyl-(1→2)]-ß-d-glucopyranoside (12), and 26-O-ß-d-glucopyranosyl-(25R)-5-en-furost-3ß,17α,22α,26-tetraol-3-O-α-l-rhamnopyranosyl-(1→4)-α-l-rhamnopyranosyl-(1→4)-[α-l-rhamnopyranosyl-(1→2)]-ß-d-glucopyranoside (13).

Ipvelutine, 7ß-Acetoxy-2α-(tigloyloxy)tropane, an Unusual Tropane Alkaloid from Ipomoea velutina R. Br. (Convolvulaceae).

Convolvulaceae provide a rich source of tropane alkaloids, however, 2-substituted tropanes have been described for only few species of this taxon. In this note, 2,7-diesters such as ipvelutine [7ß-acetoxy-2α-(tigloyloxy)tropane] isolated from the vegetative parts of the Australian Ipomoea velutina R. BR. are described as a new group of tropane diesters.

Stereochemistry of Consabatine from Convolvulus sabatius Viv. (Convolvulaceae).

The stereochemistry of consabatine, which was isolated from the roots of Convolvulus sabatius Viv. as a novel natural compound, has now been determined by the synthesis of its Mosher esters. Consabatine was found to be 1'R-configurated.

Major constituents and cytotoxic effects of Ajuga chamaecistus ssp. tomentella.

The n-butanolic fraction of a methanolic extract (80%) from aerial parts of Ajuga chamaecistus ssp. tomentella was analysed using different chromatographic methods. Column (CC) and high-performance liquid chromatography (HPLC) were used for isolation and purification. 13C, H NMR, H-H COSY, HSQC, HMBC, and ESI-MS were employed for identification of the compounds isolated from this fraction. The structures of the compounds were determined to be cis-melilotoside (1), trans-melilotoside (2), lavandulifolioside (3), 20-hydroxyecdysone (4), leonoside B (5), martynoside (6), ajugalactone (7), makisterone A (8), and 24-dehydroprecyasterone (9). This is the first report on the presence of cis- and trans-melilotoside in Ajuga species. Cytotoxic evaluation of the n-butanolic fraction, cis- and trans-melilotoside against cancer (T47D, HT-29, and Caco-2) and normal (NIH 3T3) cell lines by the mitochondrial tetrazolium test (MTT) showed no cytotoxic effects up to 400 microg/mL. The results of this study suggest that melilotoside, phenylethyl glycosides, and phytoecdysteroids are the main constituents of the n-butanolic fraction of Ajuga chamaecistus ssp. tomentella.

The toxin component of targeted anti-tumor toxins determines their efficacy increase by saponins.

Tumor-targeting protein toxins are composed of a toxic enzyme coupled to a specific cell binding domain that targets cancer-associated antigens. The anti-tumor treatment by targeted toxins is accompanied by dose-limiting side effects. The future prospects of targeted toxins for therapeutic use in humans will be determined by reduce side effects. Certain plant secondary metabolites (saponins) were shown to increase the efficacy of a particular epidermal growth factor receptor (EGFR)-targeted toxin, paralleled by a tremendous decrease of side effects. This study was conducted in order to investigate the effects of substituting different toxin moieties fused to an EGF ligand binding domain on the augmentative ability of saponins for each against therapeutic potential of the saponin-mediated efficacy increase for different anti-tumor toxins targeting the EGFR. We designed several EGFR-targeted toxins varying in the toxic moiety. Each targeted toxin was used in combination with a purified saponin (SA1641), isolated from the ornamental plant Gypsophila paniculata L. SA1641 was characterized and the SA1641-mediated efficacy increase was investigated on EGFR-transfected NIH-3T3 cells. We observed a high dependency of the SA1641-mediated efficacy increase on the nature of toxin used for the construction of the targeted toxin, indicating high specificity. Structural alignments revealed a high homology between saporin and dianthin-30, the two toxic moieties that benefit most from the combination with SA1641. We further demonstrate that SA1641 did not influence the plasma membrane permeability, indicating an intracellular interaction of SA1641 and the toxin components of targeted toxins. Surface plasmon resonance measurements point to a transient binding of SA1641 to the toxin components of targeted toxins.

Chemical composition and biological activity of Paris quadrifolia L.

A study of the components of Paris quadrifolia was undertaken to identify compounds with potential influence on cardiac cells, since previous reports suggested a cardiotoxic risk of this plant. Compounds isolated and identified included one new steroidal saponin, (23S,24S)-spirosta-5,25(27)-diene-1beta,3beta,21,23,24-pentol-1-O-beta-D-apiofuranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-xylopyranosyl-(1-->3)]-beta-D-glucopyranoside 21-O-beta-D-apiofuranoside 24-O-beta-D-fucopyranoside (1), demonstrating quite unusual structural features, as well as the known compounds 26-O-beta-D-glucopyranosyl-(25R)-5-en-furost-3beta,17alpha,22alpha,26-tetraol-3-O-alpha-L-rhamnopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->4)-[alpha-L-rhamnopyranosyl--(1-->2)]-beta-D-glucopyranoside (2), pennogenin 3-O-alpha-L-rhamnopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->4)-[alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (3), 7-O-beta-D-glucopyranosyl-kaempferol-3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-galactopyranoside (4), kaempferol-3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-galactopyranoside (5), 5-hydroxyecdysterone (6), and 20-hydroxyecdysone (7). The pennogenin derivative 3 showed strong cardiotoxic effects in an in vitro cellular model system, whereas the respective furostanol derivative 2 was inactive.

A convenient method for saponin isolation in tumour therapy.

Saponinum album (Merck), which is a crude mixture of saponins from Gypsophila paniculata L., was shown to improve the anti cancer therapy when used in vivo in combination with saporin-based targeted toxins. Unfortunately saponinum album cannot be used for further development since Merck has ceased its production in the 1990s. As pure saponins are mandatory for use in medical purposes we developed a convenient method for saponin isolation directly from the roots of Gypsophila paniculata L. The developed method is rapid, cheap and scaling up is also possible. By combining dialysis and HPLC three saponins were isolated in a one-step procedure. Chemical structures of the purified saponins were characterized by extensive one and two-dimensional NMR-spectroscopy and by using ESI-TOF-MS. The biological activities of the purified saponins were also investigated. The method presented herein enabled a rapid and cheap isolation of saponins for tumour therapy.

New cucurbitane-type triterpenoids from Bryonia aspera.

Phytochemical investigation of the chloroform extract of BRYONIA ASPERA roots resulted in the isolation and structure elucidation of two new cucurbitacins (1, 2) together with eight known cucurbitane derivatives (6-13), the pentacyclic triterpene bryonolic acid (5) and two hydroxybenzoic acid amides (3, 4). Their structures were elucidated by spectroscopic analysis, including 2D NMR techniques.

New phenolic compounds of Acmella ciliata.

The phytochemical analysis of a butanolic fraction of Acmella ciliata Kunth. afforded a total of 15 hydrophilic compounds. Seven flavonol 3-O-glycosides were isolated, including two new natural products, quercetin-3-O-(3-O-acetyl-beta-D-glucuronopyranoside) and quercetin-3-O-(2- O-acetyl-alpha-L-rhamnopyranosyl-(1--> 6)-beta-D-glucopyranoside). Furthermore, a variety of caffeoylquinic acid derivatives could be identified using ESI-MS and NMR-spectroscopy.

Extracts of Cynomorium songaricum protect SK-N-SH human neuroblastoma cells against staurosporine-induced apoptosis potentially through their radical scavenging activity.

In traditional Chinese medicine a number of herbs are used to alleviate the symptoms of aging, among them the stems of Cynomorium songaricum, Cynomoriaceae. This study evaluated the protective effect of different extracts of C. songaricum on staurosporine-induced apoptotic cell death in SK-N-SH neuroblastoma cells. Staurosporine (100 nm) reduced cell viability to about 55%. The ethyl acetate fraction of C. songaricum significantly attenuated staurosporine-induced cell death at concentrations of 100 and 10 microg/mL. On the other hand, the dichloromethane as well as water fractions showed no protective effects. In order to further analyse the protective mode of action, the superoxide anion scavenging activity of two extracts was evaluated in a xanthine/xanthine oxidase system. In this system, the EtOAc extract showed a good scavenging activity (IC(50) value 2.9 microg/mL) without inhibition of xanthine oxidase. In conclusion, the results prove the neuroprotective activity of C. songaricum extracts in vitro, thus supporting its traditional use.

Phenylethanoid glycosides from Stachytarpheta cayennensis (Rich. ) Vahl, Verbenaceae, a traditional antimalarial medicinal plant / Glicosídeos feniletanóides de Stachytarpheta cayennensis (Rich. ) Vahl, Verbenaceae, uma planta medicinal tradicional anti-malárica

The boiled juice of Stachytarpheta cayennensis, Verbenaceae, whole plant material is used ethnomedicinally in different countries of Latin-America to treat symptoms of malaria. In the ethyl acetate extract, five phenylethanoid glycosides could be identified. Three of these phenylethanoid glycosides, leucosceptoside A, martynoside and jionoside D were detected for the first time in the genus Stachytarpheta.

O suco fervido de Stachytarpheta cayennensis, Verbenaceae, do material vegetal inteiro é utilizado etnomedicinalmente em diferentes países da América Latina para tratar os sintomas da malária. No extrato acetato de etila, cinco glicosídeos feniletanóides puderam ser identificados. Três destes glicosídeos feniletanóides, leucosceptosídeo A, martinosídeo e jionosídeo D foram detectados pela primeira vez no gênero Stachytarpheta.

Plants traditionally used against malaria: phytochemical and pharmacological investigation of Momordica foetida

Leaves from Momordica foetida traditionally used to treat symptoms of malaria in parts of East Africa were studied for in vitro antimalarial activity. Using an [³H] hypoxanthine-incorporation assay the antiplasmodial activity of hydrophilic and lipophilic extracts against the chloroquine-sensitive strain poW and the multiresistant clone Dd2 of Plasmodium falciparum was determined. The petrol ether/ethyl acetate extract showed significant activity with IC50 values of 7.3 µg/mL (poW) and 13.0 µg/mL (Dd2). Cytotoxicity was estimated on two human cell lines HepG2 and ECV-304 and a selectivity index (SI) was calculated, ranging between 12 and 15. Phytochemical analysis led to the isolation of a number of phenolic glycosides, e.g. eriodictyol-, 5,7,4-trihydroxyflavanone-, kaempferol- and 5,7-dihydroxychromone-7-O-beta-D-glucopyranoside, not previously known from M. foetida. In order to prove one possible mode of action the influence of the crude extracts as well as the isolated compounds on the heme degradation with chloroquine as reference compound was evaluated. The ethyl acetate extract showed about 88 percent inhibition of heme degradation, quite similar to chloroquine (84 percent), and also eriodictyol-7-O-beta-D-glucopyranoside (2) inhibited heme degradation by 86 percent.

Folhas de Momordica foetida tradicionalmente usadas para tratar sintomas de malária em partes da Africa Oriental foram estudadas in vitro com respeito à sua atividade antimalárica. Utilizando-se o teste de incorporação de hipoxantina [³H] a atividade antiplasmódica dos extratos hidrofílico e lipofílico contra a cepa sensíveis a cloroquina poW e o clone multiresistente Dd2 de Plasmodium falciparum foi determinada. O extrato éter de petróleo/acetato de etila mostrou atividade significativa com valores de CI50 de 7,3 µg/mL (poW) e 13,0 µg/mL (Dd2). A citotoxicidade foi estimada em duas linhas de células humanas HepG2 e ECV-304 e o índice de seletividade (IS) foi calculado, variando entre 12 e 15. A análise fitoquímica levou ao isolamento de um número de glicosídeos fenólicos, por exemplo eriodictiol-, 5,7,4-triidroxiflavanona, kaempferol e 5,7-diidroxicromona-7-O-beta-D-glicopiranosideo, descritos pela primeira vez de M. foetida. Na tentativa de provar um possível modo de ação foi avaliada a influência dos extratos brutos bem como dos compostos isolados sobre a degradação da heme usando-se cloroquina como composto de referência. O extrato acetato de etila mostrou inibição da degradação da heme em torno de 88 por cento, bastante similar a cloroquina (84 por cento) e também o eriodictiol-7-O-beta-D-glicopiranosideo (2) inibiu a degradação da heme em 86 por cento.