[Separation and structural elucidation of C25 epimers of furostanol saponin from aerial parts of Paris polyphylla var. chinensis].

Paris polyphylla var. chinensis(PPC) is used as one of the origin plants of Paridis Rhizoma described in the Chinese Pharmacopoeia(2020 edition). Its resources shortage makes the planting scale gradually expand, and plenty of aerial parts are abandoned because of not being effectively used. On the basis of previous research, this study separated steroidal saponins to further clarify the chemical composition of the aerial parts of PPC. As a result, three pairs of 25R or 25S epimers of furostanol saponins were obtained by various column chromatography techniques. Their structures were identified as neosolanigroside Y6(1), solanigroside Y6(2), neoprotogracillin(3), protogracillin(4), neoprotodioscin(5) and protodioscin(6) by spectral data combining with chemical transformation. Compound 1 is a new compound, and compounds 2, 3 and 5 are isolated from Paris plants for the first time. Compounds 4 and 6 are isolated from this plant for the first time. Previously, only several spirostanol glycosides with 25S configuration were isolated from Paris plants. Guided by mass spectrometry, the present study isolated the furostanol saponins with 25S configuration from this genus for the first time, which further enriches the chemical information of Paris genus and provides a reference for the isolation of similar compounds.

Analysis of Chemical Variations between Crude and Salt-Processed Anemarrhenae rhizoma Using Ultra-High-Performance Liquid Chromatography-Mass Spectrometry Methods.

The present study was designed to systematically investigate the chemical profile differences between crude Anemarrhenae rhizoma (CAR) and salt-processed Anemarrhenae rhizoma (SAR). Ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS), coupled with multivariate statistical analysis was used for the discrimination of chemical profiles and the identification of the differentiation of the chemical constitutions of CAR and SAR. In addition, seven main constituents of CAR and SAR were simultaneously determined by ultra-high-performance liquid chromatography-quadrupole mass spectrometry (UHPLC-MS) for analyzing the content variations. A total of 24 components were found to be the main contributors to the significant difference between CAR and SAR. The structures of the marker compounds were identified based on their chromatographic behaviors, intact precursor ions, and characteristic MS fragmentation patterns. The potential structural transformation mechanism of furostanol saponins during salt processing was explored. The results may provide a scientific foundation for deeply elucidating the processing mechanism of Anemarrhenae rhizoma.

Application of high-performance countercurrent chromatography for the isolation of steroidal saponins from Liriope plathyphylla.

High-performance countercurrent chromatography (HPCCC) with electrospray light-scattering detection was applied for the first time to isolate a spirostanol and a novel furostanol saponin from Liriope platyphylla. Due to the large differences in KD values between the two compounds, a two-step HPCCC method was applied in this study. The primary HPCCC employed methylene chloride/methanol/isopropanol/water (9:6:1:4 v/v, 4 mL/min, normal-phase mode) conditions to yield a spirostanol saponin (1). After the primary HPCCC run, the solute retained in the stationary phase (SP extract) in HPCCC column was recovered and subjected to the second HPCCC on the n-hexane/n-butanol/water system (1:9:10 v/v, 5 mL/min, reversed-phase mode) to yield a novel furostanol saponin (2). The isolated spirostanol saponin was determined to be 25(S)-ruscogenin 1-O-ß-D-glucopyranosyl (1→2)-[ß-D-xylopyranosyl (1→3)]-ß-D-fucopyranoside (spicatoside A), and the novel furostanol saponin was elucidated to be 26-O-ß-D-glucopyranosyl-25(S)-furost-5(6)-ene-1ß-3ß-22α-26-tetraol-1-O-ß-D-glucopyranosyl (1→2)-[ß-D-xylopyranosyl-(1→3)]-ß-D-fucopyranoside (spicatoside D).

New steroidal glycosides from Hosta plantaginea (Lam.) Aschers.

Four new furostanol glycosides were isolated from the flowers of Hosta plantaginea (Lam.) Aschers. On the basis of spectroscopic methods including 1D and 2D NMR experiments, their structures were elucidated as 26-O-ß-d-glucopyranosyl-(25R)-22-O-methyl-5α-furostan-2α,3ß,22ξ,26-tetrol 3-O-α-l-rhamnopyranosyl-(1 â†’ 4)-O-ß-d-xylopyranosyl-(1 â†’ 3)-[O-ß-d-glucopyranosyl-(1 â†’ 2)]-O-ß-d-glucopyranosyl-(1 â†’ 4)-ß-d-galactopyranoside (hostaplantagineoside A, 1), 26-O-ß-d-glucopyranosyl-(25R)-5α-furostan-20(22)-ene-2α,3ß,26-triol-3-O-ß-d-glucopyranosyl-(1 â†’ 2)-[O-ß-d-xylopyranosyl-(1 â†’ 3)]-O-ß-d-glucopyranosyl-(1 â†’ 4)-ß-d-galactopyranoside (hostaplantagineoside B, 2), 26-O-ß-d-glucopyranosyl-(25R)-5α-furostan-22(23)-ene-2α,3ß,20α,26-tetraol-3-O-ß-d-glucopyranosyl-(1 â†’ 2)-[O-ß-d-xylopyranosyl-(1 â†’ 3)]-O-ß-d-glucopyranosyl-(1 â†’ 4)-O-ß-d-galactopyranoside (hostaplantagineoside C, 3), 26-O-ß-d-glucopyranosyl-(25R)-5α-furostan-20(22)-ene-2α,3ß,26-triol-3-O-α-l-rhamnopyranosyl-(1 â†’ 4)-O-ß-d-xylopyranosyl-(1 â†’ 3)-[O-ß-d-glucopyranosyl-(1 â†’ 2)]-O-ß-d-glucopyranosyl-(1 â†’ 4)-ß-d-galactopyranoside (hostaplantagineoside D, 4).

Steroidal glycosides with antiproliferative activities from Digitalis trojana.

The phytochemical investigation of Digitalis trojana led to the isolation of two cardiac glycosides (1, 2), one pregnane glycoside (3), three furostanol type saponins (4-6), along with three cleroindicins (7-9), four phenylethanoid glycosides (10-13), two flavonoids (14, 15) and two phenolic acid derivatives (16, 17). The structure elucidation of the isolates was carried out by NMR experiments as well as ESI-MS. The cytotoxic activity of compounds 1-13 against a small panel of cancer cell lines, namely MCF-7, T98G, HT-29, PC-3, A375 and SH-SY5Y, was investigated. Compounds 1-6 showed antiproliferative activity against human breast MCF-7 and colon HT-29 cancer cell lines with IC50 values ranging from 8.3 to 50 µM. In order to understand the mechanism involved in the cell death, the active compounds were tested as pro-apoptotic agents using propidium iodide staining by flow cytometry method. No significant increase was observed in the apoptosis of the MCF-7 and HT-29 cancer cells. Moreover, the effects of the active compounds on cell proliferation were assessed on the same cancer cell lines by cell cycle analysis of DNA content using flow cytometry. No significative changes were observed in the cell cycle of MCF-7, while significant changes in G2 /M cell cycle phase of HT-29 cells were observed after treatment with digitalin (1), cariensoside (3) and 22-O-methylparvispinoside B (6) at 10 µM.

Cytotoxic steroidal glycosides from the rhizomes of Paris polyphylla var. yunnanensis.

Paris polyphylla var. yunnanensis (Franch.) Hand.-Mazz. (Melanthiaceae), an important specie of the genus Paris, has long been in a traditional Chinese medicine (TCM) for a long time. This study aimed to isolate and identify the structures of bioactive saponins from the rhizomes of P. polyphylla var. yunnanensis and evaluate their cytotoxicity against BxPC-3, HepG2, U373 and SGC-7901 carcinoma cell lines. Seven previously undescribed and seven known saponins were identified, and Paris saponins VII (PSVII) showed significant cytotoxicity against the BxPC-3 cell line with IC50 values of 3.59 µM. Furthermore, flow cytometry, transmission electron microscopy and western-bolt analysis revealed that PSVII inhibited the proliferation of BxPC-3 cells and might be involved in inducing apoptosis and pyroptosis by activating caspase-3, -7 and caspase-1, respectively.

Parisvaniosides A-E, five new steroidal saponins from Paris vaniotii.

The species of Paris genus is a prolific source of structurally diverse steroidal saponins responsible for multivarious biological properties. The first phytochemical investigation on the steroidal saponin constituents from the rhizomes of Paris vaniotii Lévl. led to the discovery and structural characterization of four new spirostanol saponins, named parisvaniosides A-D (1-4), and one new furostanol glycoside, named parisvanioside E (5), along with eleven known analogues (6-16). Their structures were unambiguously established on the basis of extensive spectroscopic analysis and comparison with the reported spectroscopic data. Compound 1 is a rare spirostanol saponin sharing with a C-9/C-11 double bond and a peroxy group located between C-5 and C-8 of the aglycone, whereas 3 and 4 are unusual C-27 steroidal sapoins with hydroxyl/methoxyl at both C-5 and C-6. Furthermore, 5 is the first furostanol saponin with a unique aglycone featuring two trisubstituted double bonds in ring B. All isolated saponins were evaluated for their anti-inflammatory effects on a lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production model in RAW 264.7 macrophages.

Five new polyhydroxylated furostanol saponins from the rhizomes of Tupistra chinensis.

Five new polyhydroxylated furostanol saponins were isolated from the roots and rhizomes of Tupistra chinensis, and their structures were determined as tupistrosides J-N (1-5), together with four known furostanol saponins (6-9), on the basis of physico-chemical properties and spectral analysis. Among them, compounds 3 and 5 showed cytotoxicity against human cancer cell lines SW620 with IC50 values of 72.5 ± 2.4 and 77.3 ± 2.5 µmol·L-1, respectively. Compound 4 showed cytotoxicity against human cancer cell line HepG2 with IC50 value of 88.6 ± 2.1 µmol·L-1.

Furostanol saponins from Chinese onion induce G2/M cell-cycle arrest and apoptosis through mitochondria-mediate pathway in HepG2 cells.

Phytochemical investigations on the bulbs of Chinese onion led to the isolation of three new furostanol saponins (1, 2, 5) together with seven known furostanol saponins (3, 4, 6-10). Their chemical structures were elucidated on the basis of spectroscopic and chemical methods, including IR, MS, NMR, and GC analyses. The anti-proliferative and anti-inflammatory activities of the isolates were evaluated. Compounds 7-10 showed potential anti-proliferative activities against human cancer cell lines (HepG2, A549, SPC-A-1, MGC80-3, MDA-MB-231, SW620 and CNE-1) with IC50 values below 30 µM. Compounds 4 and 7 could induce G2/M cell-cycle arrest and apoptosis through mitochondria-mediate pathway in HepG2 cells. Compounds 7 and 10 showed strong inhibitory effects against LPS induced NO production in RAW264.7 cells with IC50 values of 2.01 ±â€¯1.40 µM and 2.49 ±â€¯1.54 µM, respectively.

New furostanol saponins with anti-inflammatory and cytotoxic activities from the rhizomes of Smilax davidiana.

Seven new furostanol saponins have been isolated from the rhizomes of Smilax davidiana. Their structures were established by 2D NMR spectroscopic techniques (1H,1H-COSY, NOESY, HSQC and HMBC), mass spectrometry and comparison with the literature. The isolated compounds were subjected to evaluate anti-inflammatory and cytotoxic activities in vitro. Compounds 3, 5 and 7 were found to have modest anti-inflammatory effects through suppression of IL-1ß production and promote the expression of IL-10 in LPS-stimulated RAW 264.7 cells. Davidianoside F (6) showed activity against MCF-7 and HELA cell lines at the concentration of 10.2µM and 4.3µM, respectively.

Antiproliferative and anti-inflammatory furostanol saponins from the rhizomes of Tupistra chinensis.

Phytochemical investigations of the rhizome of Tupistra chinensis led to the isolation of ten new furostanol saponins along with fourteen known spirostanols. Their chemical structures were elucidated on the basis of spectroscopic and chemical methods, including IR, NMR, MS, and GC analyses. The antiproliferative effects against FaDu and Detroit 562 cell lines and inhibitory activities on nitric oxide (NO) production induced by lipopolysaccharide (LPS) in a macrophage cell line RAW 264.7 were assayed for all the isolated compounds. Compound 14 exhibited significant antiproliferative effects against FaDu and Detroit 562 cells with IC50 values of 1.1±0.1 and 1.2±0.1µM, respectively. Compounds 1, 2, 6, 13, 16, 19 and 24 exhibited inhibitory effects on NO production with IC50 values ranging from 15.7 to 46.2µM.

Furostanol saponins from the seeds of Allium cepa L.

Allium cepa L. is one of the most widely cultivated and used plants. In addition to its bulb (onion), which is used as food in many cultures, the seeds of A. cepa L. are used as a traditional herbal medicine by the Uygur nationality in China to treat diarrhea and promote blood flow. In a bioactivity-screening, the ethanol extract of seeds of A. cepa L. showed inhibitory effects on protein tyrosine phosphatase 1B (PTP1B) enzyme, with 81.1% inhibition. Phytochemical investigation of the ethanol extract of red onion (Allium cepa L.) seeds led to the isolation of eight new furostanol saponins, named ceparosides E-L (1-8). Their structures were established using 1D and 2D NMR spectroscopy, mass spectrometry and chemical methods. Compounds 1-8 were screened for inhibitory effects on the PTP1B enzyme and cytotoxic activity against five human cells, including HCT-8, Bel-7402, BGC-823, A549 and A2780, but all were found to be inactive.