Anti-inflammatory steroidal sapogenins and a conjugated chalcone-stilbene from Dracaena usambarensis Engl.

Four new steroidal sapogenins, dracaenogenins CF (1-4), a new conjugated chalcone-stilbene, 3''-methoxycochinchinenene H (5) together with eight known compounds namely, (25S)-spirosta-1,4-dien-3-one (6), trans-resveratrol (7), 4,4'-dihydroxy-3'-methoxychalcone (8), N-trans-coumaroyltyramine (9), N-trans-p-coumaroyloctopamine (10), N-trans-feruloyloctopamine (11), 7-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-N2,N3-bis(4-hydroxyphenethyl)-6-methoxy-1,2-dihydronaphthalene-2,3-dicarboxamide (12) and grossamide (13) were isolated from the stems of Dracaena usambarensis Engl. from Kenya. It is important to note that compounds 12 and 13 are being reported from this genus for the first time. Structural elucidation of the isolated compounds was done using spectroscopic (NMR, UV, IR, optical rotation) and spectrometric (HRESIMS) techniques. The absolute and relative configurations of the isolated compounds were determined by employing single crystal X-ray crystallography analysis, NOESY correlations and coupling constants. The anti-inflammatory potencies of the isolated compounds were evaluated by measuring the levels of four cytokines (IL-1ß, IL-2, GM-CSF and TNF-α) in the supernatant media of human peripheral blood mononuclear cells (PBMCs) stimulated by lipopolysaccharide (LPS). At the tested concentration of 100 µM, the new conjugated chalcone-stilbene 5, the dihydrochalcone, 8 and the lignanamide, 13 were substantially more potent than the standard drug, ibuprofen, inhibiting the release of all the cytokines, IL-1ß, IL-2, GM-CSF and TNF-α from 0.06-58.04% compared to LPS control. These compounds should therefore be considered for development into anti-inflammatory drug candidates. Compound 7 significantly decreased the release of GM-CSF (6.11% of LPS control) and TNF-α (18.35% of LPS control). The cytokine TNF-α was sensitive to all the tested compounds 1-13.

Distribution of sapogenins in morphological Medicago sativa L. parts: Comparison of various extraction techniques.

Saponins in plant extracts were indirectly determined by estimation of the content of sapogenins. The first step of determination is extraction with high efficiency. One conventional extraction technique (maceration) and two modern ones (accelerated solvent extraction and supercritical fluid extraction) were compared. Methanol and ethanol were used as solvents or co-solvents. The results were supported by statistical analysis. Saponins were extracted from leaves, roots, and sprouts of Medicago sativa. Acid hydrolysis, purification, and determination by high-performance liquid chromatography with evaporative light scattering detector were used. The content of sapogenins was the highest in the roots. Smaller amounts of sapogenins were found in sprouts and the smallest ones in leaves. The main ingredient was medicagenic acid with mean concentration of 621.8 µg/g in roots, 456.7 µg/g in sprouts, and 471.3 µg/g in leaf extract. The highest content of sapogenins in extract was obtained after maceration with methanol; however, this method is nonselective in relation to biologically active compounds. Due to the possibility of using the obtained extracts with sapogenins in the cosmetic or pharmaceutical industry, the selection of extraction techniques and solvents is a very important aspect. Additionally, the chosen technique should be considered eco-friendly and consistent with the assumptions of "green chemistry."

Cycloastragenol ameliorates experimental heart damage in rats by promoting myocardial autophagy via inhibition of AKT1-RPS6KB1 signaling.

Cycloastragenol, a naturally occurring compound in Astragali Radix, has been demonstrated to possess various pharmacological actions including anti-aging, anti-inflammation, anti-fibrosis, antibacterial, liver and endothelium protection. However, whether cycloastragenol ameliorates heart failure remains unclear. Isoproterenol administration to rats triggered classic cardiac damage, as demonstrated by objective parameters of cardiac dysfunction. The treatment of cycloastragenol improved deranged cardiac parameters in the isoproterenol-induced heart damage model in a dose-dependent manner. At the same time, cycloastragenol markedly ameliorated cardiac histological changes and down-regulated serum levels of various neuroendocrine factors including norepinephrine, aldosterone, brain natriuretic peptide, endothelin 1, angiotensin II and so on. Moreover, the expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 in rat heart were also inhibited by cycloastragenol. Mechanistically, augmenting autophagy of myocardial cells via the inhibition of AKT1-RPS6KB1 signaling contributed to the improvement of isoproterenol-induced rat heart failure by cycloastragenol. These results suggest that cycloastragenol ameliorates cardiac dysfunction and remodeling through promoting autophagy in myocardial cells and suppressing MMP-2 and MMP-9 expressions, indicating that it could be a drug candidate for patients with congestive heart failure.

Bupleurum chinense Roots: a Bioactivity-Guided Approach toward Saponin-Type NF-κB Inhibitors.

The roots of Bupleurum chinense have a long history in traditional medicine to treat infectious diseases and inflammatory disorders. Two major compounds, saikosaponins A and D, were reported to exert potent anti-inflammatory activity by inhibiting NF-κB. In the present study, we isolated new saikosaponin analogues from the roots of B. chinese interfering with NF-κB activity in vitro. The methanol-soluble fraction of the dichloromethane extract of Radix Bupleuri was subjected to activity-guided isolation yielding 18 compounds, including triterpenoids and polyacetylenes. Their structures were determined by spectroscopic methods as saikogenin D (1), prosaikogenin D (2), saikosaponins B2 (3), W (4), B1 (5), Y (6), D (7), A (8), E (9), B4 (10), B3 (11), and T (12), saikodiyne A (13), D (14), E (15) and F (16), falcarindiol (17), and 1-linoleoyl-sn-glycero-3-phosphorylcholine (18). Among them, 4, 15, and 16 are new compounds, whereas 6, previously described as a semi-synthetic compound, is isolated from a natural source for the first time, and 13-17 are the first reports of polyacetylenes from this plant. Nine saponins/triterpenoids were tested for inhibition of NF-κB signaling in a cell-based NF-κB-dependent luciferase reporter gene model in vitro. Five of them (1, 2, 4, 6, and 8) showed strong (> 50%, at 30 µM) NF-κB inhibition, but also varying degrees of cytotoxicity, with compounds 1 and 4 (showing no significant cytotoxicity) presenting IC50 values of 14.0 µM and 14.1 µM in the cell-based assay, respectively.

Synthesis and neuroprotective effects of the complex nanoparticles of iron and sapogenin isolated from the defatted seeds of Camellia oleifera.

CONTEXT: The defatted seeds of Camellia oleifera var. monosperma Hung T. Chang (Theaceae) are currently discarded without effective utilization. However, sapogenin has been isolated and shows antioxidative, anti-inflammatory and analgesic activities suggestive of its neuroprotective function. OBJECTIVE: In order to improve the activities of sapogenin, the nanoparticles of iron-sapogenin have been synthesized, and the neuroprotective effects are evaluated. MATERIALS AND METHODS: Structural characters of the nanoparticles were analyzed, and the antioxidant effect was assessed by DPPH method, and the neuroprotective effect was evaluated by rotenone-induced neurodegeneration in Kunming mice injected subcutaneously into the back of neck with rotenone (50 mg/kg/day) for 6 weeks and then treated by tail intravenous injection with the iron-sapogenin at the dose of 25, 50 and 100 mg/kg for 7 days. Mice behaviour and neurotransmitters were tested. RESULTS: The product had an average size of 162 nm with spherical shape, and scavenged more than 90% DPPH radicals at 0.8 mg/mL concentration. It decreased behavioural disorder and malondialdehyde content in mice brain, and increased superoxide dismutase activity, tyrosine hydroxylase expression, dopamine and acetylcholine levels in brain in dose dependence, and their maximum changes were respectively up to 60.83%, 25.17%, 22.13%, 105.26%, 42.17% and 22.89% as compared to vehicle group. Iron-sapogenin nanoparticle shows significantly better effects than the sapogenin. DISCUSSION AND CONCLUSION: Iron-sapogenin alleviates neurodegeneration of mice injured by neurotoxicity of rotenone, it is a superior candidate of drugs for neuroprotection.

Cross Interaction Between Ilyonectria mors-panacis Isolates Infecting Korean Ginseng and Ginseng Saponins in Correlation with Their Pathogenicity.

Ilyonectria mors-panacis belongs to I. radicicola species complex and causes root rot and replant failure of ginseng in Asia and North America. The aims of this work were to identify I. mors-panacis that infect Korean ginseng using molecular approaches and to investigate whether their aggressiveness depends on their ability to metabolize ginseng saponins (ginsenosides) by their ß-glucosidases, in comparison with other identified Ilyonectria species. Fourteen isolates were collected from culture collections or directly isolated from infected roots and mainly identified based on histone H3 (HIS H3) sequence. Among them, six isolates were identified as I. mors-panacis while others were identified as I. robusta and I. leucospermi. The pathogenicity tests confirmed that the isolates of I. mors-panacis were significantly more aggressive than I. robusta and I. leucospermi. The major ginsenosides in I. mors-panacis-infected roots were significantly reduced while significantly increased in those infected with other species. In vitro, the isolates were tested for their sensitivity and ability to metabolize the total major ginsenosides (Total MaG), protopanaxadiol-type major ginsenosides (PPD-type MaG), and protopanaxatriol-type major ginsenosides (PPT-type MaG). Unexpectedly, the growth rate and metabolic ability of I. mors-panacis isolates were significantly low on the three different ginsenoside fractions while those of I. robusta and I. leucospermi were significantly reduced on PPT-type MaG and Total MaG fractions and not affected on PPD-type MaG fraction. Our results indicate that major ginsenosides, especially PPT-type, have an antifungal effect and may intervene in ginseng defense during Ilyonectria species invasion, in particular the weak species. Also, the pathogenicity of I. mors-panacis may rely on its ability to reduce saponin content; however, whether this reduction is caused by detoxification or another method remains unclear.

Fermented Ginseng Contains an Agonist of Peroxisome Proliferator Activated Receptors α and γ.

Peroxisome proliferator activated receptor (PPAR) is a nuclear receptor that is one of the transcription factors regulating lipid and glucose metabolism. Fermented ginseng (FG) is a ginseng fermented by Lactobacillus paracasei A221 containing minor ginsenosides and metabolites of fermentation. DNA microarray analysis of rat liver treated with FG indicated that FG affects on lipid metabolism are mediated by PPAR-α. To identify a PPAR-α agonist in FG, PPAR-α transcription reporter assay-guided fractionation was performed. The fraction obtained from the MeOH extract of FG, which showed potent transcription activity of PPAR-α, was fractionated by silica gel column chromatography into 16 subfractions, and further separation and crystallization gave compound 1 together with four known constituents of ginseng, including 20(R)- and 20(S)-protopanaxadiol, and 20(R)- and 20(S)-ginsenoside Rh1. The structure of compound 1 was identified as 10-hydroxy-octadecanoic acid by (1)H- and (13)C-NMR spectra and by EI-MS analysis of the methyl ester of 1. Compound 1 demonstrated much higher transcription activity of PPAR-α than the other isolated compounds. In addition, compound 1 also showed 5.5-fold higher transcription activity of PPAR-γ than vehicle at the dose of 20 µg/mL. In the present study, we identified 10-hydroxy-octadecanoic acid as a dual PPAR-α/γ agonist in FG. Our study suggested that metabolites of fermentation, in addition to ginsenosides, contribute to the health benefits of FG.

Tartaric acid induced conversion of protopanaxadiol to ginsenosides Rg3 and Rg5 and their in situ recoveries by integrated expanded bed adsorption chromatography.

Panax ginseng has been applied in traditional Chinese medicine for over 2000 years. It is still one of the most popular herbs in recent decades. The prescribed ginseng-containing medicines consist of protopanaxadiol and protopanaxatriol ginsenosides, which are the major constituents of the herb. Minor ginsenosides at low levels in the herb, such as Rg3 and Rg5 , have attracted more rising attention than the major ones. The existing approaches to prepare Rg3 and Rg5 usually rely on either steamed red ginseng as the source or chemical/enzymatic conversion of protopanaxadiol to the targets. It is still highly desirable to effectively achieve such minor components. In this paper, a method integrated extraction of protopanaxadiol and conversion of it to Rg3 and Rg5 has been proposed. Protopanaxadiol was extracted and simultaneously converted to Rg3 and Rg5 by d,l-tartaric acid. The targets were absorbed by resins on expanded bed adsorption chromatography and were then separated from other ginsenosides in different stages. Compared with conventional methods, the developed process has advantages in shortening time consumption and improving the conversion ratio of protopanaxadiol, which is promising in directly achieving Rg3 and Rg5 from P. ginseng.

Phytochemistry and Anticancer Potential of Notoginseng.

Asian ginseng, American ginseng, and notoginseng are three major species in the ginseng family. Notoginseng is a Chinese herbal medicine with a long history of use in many Oriental countries. This botanical has a distinct ginsenoside profile compared to other ginseng herbs. As a saponin-rich plant, notoginseng could be a good candidate for cancer chemoprevention. However, to date, only relatively limited anticancer studies have been conducted on notoginseng. In this paper, after reviewing its anticancer data, phytochemical isolation and analysis of notoginseng is presented in comparison with Asian ginseng and American ginseng. Over 80 dammarane saponins have been isolated and elucidated from different plant parts of notoginseng, most of them belonging to protopanaxadiol or protopanaxatriol groups. The role of the enteric microbiome in mediating notoginseng metabolism, bioavailability, and pharmacological actions are discussed. Emphasis has been placed on the identification and isolation of enteric microbiome-generated notoginseng metabolites. Future investigations should provide key insights into notoginseng's bioactive metabolites as clinically valuable anticancer compounds.

Bioactive cyclolanstane-type saponins from the stems of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao.

Twelve cyclolanstane-type saponins including six new ones, astrolanosaponins A1 (1), A2 (2), B (3), C (4), D (5), and E (6) were obtained from the stem of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao, and their structures were elucidated by chemical and spectroscopic methods. Of the known ones, cycloastragenol-3-O-ß-D-glucopyranoside (7), astraverrucin II (8), cycloaraloside E (9), huangqiyenin A (10), and huangqiyenin B (11) were isolated from the species first. Meanwhile, compounds 1-3, 5-9, and aleksandroside I (12) showed inhibitory effects on triglyceride accumulation in HepG2 cells.

Fast Centrifugal Partition Chromatography Fractionation of Concentrated Agave (Agave salmiana) Sap to Obtain Saponins with Apoptotic Effect on Colon Cancer Cells.

Separation of potentially bioactive components from foods and plant extracts is one of the main challenges for their study. Centrifugal partition chromatography has been a successful technique for the screening and identification of molecules with bioactive potential, such as steroidal saponins. Agave is a source of steroidal saponins with anticancer potential, though the activity of these compounds in concentrated agave sap has not been yet explored. In this study, fast centrifugal partition chromatography (FCPC) was used coupled with in vitro tests on HT-29 cells as a screening procedure to identify apoptotic saponins from an acetonic extract of concentrated agave sap. The three most bioactive fractions obtained by FCPC at partition coefficients between 0.23 and 0.4 contained steroidal saponins, predominantly magueyoside b. Flow cytometry analysis determined that the fraction rich in kammogenin and manogenin glycosides induced apoptosis, but when gentrogenin and hecogenin glycosides were also found in the fraction, a necrotic effect was observed. In conclusion, this study provides the evidence that steroidal saponins in concentrated agave sap were potential inductors of apoptosis and that it was possible to separate them using fast centrifugal partition chromatography.

Chemical composition and medicinal significance of Fagonia cretica: a review.

Members of the family Zygophyllaceae are distributed in arid areas of the world and are traditionally used against various health insults ranging from skin lesions to lethal cancer. Fagonia cretica Linn. is a plant having novel compounds responsive in diseases that are still considered as incurable or are curable with serious side effects. Researchers, particularly of the Asian region elaborately studied the chemical composition and pharmacological activities of this plant. But further studies are still required to evaluate this plant in clinical trials in order to save humanity from synthetic chemical drugs yet disputed as 'friends or foe'.

Steroidal sapogenins and glycosides from the fibrous roots of Ophiopogon japonicus and Liriope spicata var. prolifera with anti-inflammatory activity.

Two new steroidal glycosides (1 and 2), together with 15 known compounds (3-17) were isolated from the fibrous roots of Ophiopogon japonicus, and three new steroidal glycosides (18-20), together with 14 known compounds (21-34) were isolated from the fibrous roots of Liriope spicata var. prolifera. The structures of the new compounds were elucidated on the basis of extensive one-dimensional (1D)- and 2D-NMR spectroscopic analyses and mass spectrometry. The isolated compounds were evaluated for their anti-inflammatory activity in vitro. Most of these steroidal glycosides showed significant inhibitory activity against neutrophil respiratory burst stimulated by phorbol myristate acetate.

TRAIL pathway is associated with inhibition of colon cancer by protopanaxadiol.

Among important components of American ginseng, protopanaxadiol (PPD) showed more active anticancer potential than other triterpenoid saponins. In this study, we determined the in vivo effects of PPD in a mouse cancer model first. Then, using human colorectal cancer cell lines, we observed significant cancer cell growth inhibition by promoting G1 cell cycle redistribution and apoptosis. Subsequently, we characterized the downstream genes targeted by PPD in HCT-116 cancer cells. Using Affymetrix high density GeneChips, we obtained the gene expression profile of the cells. Microarray data indicated that the expression levels of 76 genes were changed over two-fold after PPD, of which 52 were upregulated while the remaining 24 were downregulated. Ingenuity pathway analysis of top functions affected was carried out. Data suggested that by regulating the interactions between p53 and DR4/DR5, the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) pathway played a key role in the action of PPD, a promising colon cancer inhibitory compound.

3-Keto umbilicagenin A and B, new sapogenins from Allium umbilicatum Boiss.

Two sapogenins, named 3-keto umbilicagenin A and B (1 and 2), possessing a novel chemical structure with a 3-keto group on the spirostane skeleton, have been isolated from Allium umbilicatum Boiss. Their chemical structure has been established through a combination of extensive spectroscopic analysis, mainly nuclear magnetic resonance and mass spectrometry, and chemical methods as (25R)-3-keto-spirostan-2α,5α,6ß-triol (1) and (25R)-3-keto-spirostan-2α,5α-diol (2). The isolated compounds were tested for cytotoxic activity on J-774, murine monocyte/macrophage, and WEHI-164, murine fibrosarcoma cell lines.

A novel strategy for rapid quantification of 20(S)-protopanaxatriol and 20(S)-protopanaxadiol saponins in Panax notoginseng P. ginseng and P. quinquefolium.

A novel strategy for the qualitative and quantitative determination of 20(S)-protopanaxatriol saponins (PTS) and 20(S)-protopanaxadiol saponins (PDS) in Panax notoginseng, Panax ginseng and Panax quinquefolium, based on the overlapping peaks of main components of PTS (calibrated by ginsenoside Rg1) and PDS (calibrated by ginsenoside Rb1), was proposed. The analysis was performed by using high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD). Under specific chromatographic conditions, all samples showed two overlapping peaks containing several main ginsenosides belonging to PTS and PDS, respectively. The overlapping peaks were also identified by using HPLC-MS. Based on the sum and ratio of PTS and PDS, 60 tested Panax samples were divided into three main clusters according to their species. The findings suggested that this strategy provides a simple and rapid approach to quantify PTS and PDS in Panax herbs.

Studies on cytotoxic pregnane sapogenins from Cynanchum wilfordii.

To investigate their cytotoxicity, seventeen C21-steroidal pregnane sapogenins 1-17 were isolated from the hydrolytic extracts of the roots of Cynanchum wilfordii. Among them, sapogenins 1-7 are new compounds, whose structures were determined by extensive analysis of spectroscopic data and X-ray crystallographic analysis. Especially, sapogenins with salicyl or vanilloyl group, and a new aberrant pregnane skeleton with ether linkage between C-12 and C-20 were found for the first time. Compound 1 revealed significant cytotoxicities on HL-60 (IC50 6.72µM) and MCF-7 (IC50 2.89µM), and compounds 14 and 15 also revealed strong inhibitory activities against K-562 (IC50 6.72µM) or MCF-7 (IC50 2.49µM), respectively.

Natural products and chemotherapeutic agents on cancer: prevention vs. treatment.

Natural products play an important role in cancer therapeutics, and lately more attentions have been paid to the prevention of major lethal malignancies, such as colorectal cancer (CRC). After oral ingestion, botanicals' parent compounds can be converted to their metabolites by the enteric microbiome, and these metabolites may have different bioactivities and variable bioavailability. In this study, we used an active ginseng metabolite, protopanaxadiol (PPD), as an example to assess its colon cancer preventive effect by comparing its effect with the treatment effect of fluorouracil (5-FU). A xenograft tumor nude mouse model with human colon cancer cell inoculation was used. After preventive PPD or treatment 5-FU administration with the same dose (30 mg/kg), tumor growth inhibition was evaluated by both a Xenogen bioluminescence imaging technique and manual tumor size measurement. Our data showed that preventive PPD very significantly inhibited the tumor growth compared to 5-FU (p < 0.01). Our data suggest that the PPD is a promising cancer prevention agent. More studies are needed to explore the chemopreventive actions of PPD and its potential clinical utility.

Steroidal sapogenins and glycosides from the fibrous roots of Polygonatum odoratum with inhibitory effect on tissue factor (TF) procoagulant activity.

Six new spirostane glycosides (1-6), named polygodosides A-F, one new furostanol glycoside, polygodoside G (7), one new cholestane glycoside, polygodoside H (8), and one new steroidal sapogenin, polygodosin A (9), together with thirteen known compounds (10-22) were isolated from a 90% MeOH extract of the fibrous roots of Polygonatum odoratum (Mill.) Druce. The structures of new compounds were elucidated by extensive 1D and 2D NMR spectroscopic analyses and mass spectrometry. The effects on TF procoagulant activity in THP-1 cells were tested for most of the compounds.

Smith degradation, an efficient method for the preparation of cycloastragenol from astragaloside IV.

Cycloastragenol (CA) is the genuine sapogenin of astragaloside IV (ASI). This study focuses on the preparation of CA from ASI. Five hydrolysis methods were compared including H2SO4 hydrolysis, HCl hydrolysis, two-phase acid hydrolysis, mild acid hydrolysis, and Smith degradation. Seven hydrolysis products were purified, and five of them were identified as new compounds. The results indicated that Smith degradation was the most effective approach to prepare CA. In contrast, mild acid hydrolysis produced CA at a low yield, accompanied with the artificial sapogenin astragenol. The other three acid hydrolysis methods mainly produced astragenol. Furthermore, the reaction conditions for Smith degradation were optimized as follows: ASI was dissolved in 60% MeOH-H2O solution, oxidized with 5 equiv. NaIO4 for 12h, followed by reduction with 3 equiv. NaBH4 for 4h, and finally acidified with 1M H2SO4 at pH2 for 24h. Under the optimal conditions, CA could be prepared from ASI at a yield of 84.4%.