Stilbene Oligomers Research Articles

Targeting of Antifungal Metabolites from Grapevine Byproducts by UPLC-HRMS/MS Approaches Using Bioactivity-Based Molecular Networking.

This study focuses on countering Fusarium graminearum, a harmful fungal pathogen impacting cereal crops and human health through mycotoxin production. These mycotoxins, categorized as type B trichothecenes, pose significant health risks. Research explores natural alternatives to synthetic fungicides, particularly investigating phenolics in grapevine byproducts. Thirteen eco-extracts from five French grape varieties (Merlot, Cabernet Sauvignon, Sauvignon blanc, Tannat, and Artaban) exhibited substantial antifungal properties, with ten extracts displaying remarkable effects. Extracts from grapevine stems and roots notably reduced fungal growth by over 91% after five days. Through UHPLC-HRMS/MS analysis and metabolomics, the study identified potent antifungal compounds such as ampelopsin A and cyphostemmin B, among other oligomeric stilbenes. Interestingly, this approach showed that flavan-3-ols have been identified as markers for extracts that induce fungal growth. Root extracts from rootstocks, rich in oligostilbenes, demonstrated the highest antifungal activity. This research underscores grapevine byproducts' potential both as a sustainable approach to control F. graminearum and mycotoxin contamination in cereal crops and the presence of different metabolites from the cultivars of grapevine, suggesting different activities.

Temperature affects organic acid, terpene and stilbene metabolisms in wine grapes during postharvest dehydration.

The partial dehydration of grapes after harvest is a traditional practice in several winegrowing regions that leads to the production of high quality wines. Postharvest dehydration (also known as withering) has a significant impact on the overall metabolism and physiology of the berry, yielding a final product that is richer in sugars, solutes, and aroma compounds. These changes are, at least in part, the result of a stress response, which is controlled at transcriptional level, and are highly dependent on the grape water loss kinetics and the environmental parameters of the facility where grapes are stored to wither. However, it is difficult to separate the effects driven by each single environmental factor from those of the dehydration rate, especially discerning the effect of temperature that greatly affects the water loss kinetics. To define the temperature influence on grape physiology and composition during postharvest dehydration, the withering of the red-skin grape cultivar Corvina (Vitis vinifera) was studied in two conditioned rooms set at distinct temperatures and at varying relative humidity to maintain an equal grape water loss rate. The effect of temperature was also studied by withering the grapes in two unconditioned facilities located in geographic areas with divergent climates. Technological, LC-MS and GC-MS analyses revealed higher levels of organic acids, flavonols, terpenes and cis- and trans-resveratrol in the grapes withered at lower temperature conditions, whereas higher concentrations of oligomeric stilbenes were found in the grapes stored at higher temperatures. Lower expression of the malate dehydrogenase and laccase, while higher expression of the phenylalanine ammonia-lyase, stilbene synthase and terpene synthase genes were detected in the grapes withered at lower temperatures. Our findings provide insights into the importance of the temperature in postharvest withering and its effect on the metabolism of the grapes and on the quality of the derived wines.

Production of high molecular-ordered stilbene oligomers for the study of their biological activity: total synthesis, bio-catalyzed synthesis and production by plant systems.

Though the iconic stilbene resveratrol and its related dimers constitute a top storyline in the field of natural product research, resveratrol oligomers (condensation >2) have been left aside despite their higher biological activity compared to that of the monomers. This situation largely results from the difficulty of getting them in sufficient quantities to enable evaluation of their biological properties in vivo. We present here a synthetic and critical analysis of the methods used for the production of high molecular-ordered stilbene oligomers of potential biomedical interest, gathering the most salient data regarding the approaches employed to prepare them by total synthesis, use of biomimetic approaches or through plant systems.

Identification and structural elucidation of bioactive compounds from Scirpoides holoschoenus

Phytochemical investigations of dichloromethane and methanol extracts of roots and rhizomes of Scirpoides holoschoenus afforded 21 stilbenes, six flavonoids, six ferulic acid derivatives and four diterpenes. Among these constituents, six stilbenes, one flavonoid, one diterpene and two ferulic acid derivatives, represent previously unreported natural products. Structure elucidation was performed by HRESI-MS, NMR, GC-MS, and ECD data evaluation. The monoprenylated flavonoid (sophoraflavanone B) and all isolated stilbene oligomers (trans-scirpusin B, scirpusin A, cassigarol E, cyperusphenol B, cyperusphenol D, passiflorinol A, cyperusphenol A and mesocyperusphenol A) showed strong inhibitory activities on spore germination of two Botrytis cinerea strains isolated from field-infected grape berries and apple fruits compared to the reference controls resveratrol, piceid, and fenhexamid at a test concentration of 2.0 mM. For sophoraflavanone B and cyperusphenol A, the EC50 values were determined by concentration response curves and resulted in values of 0.35 mM and 0.53 mM, respectively. The data suggest that stilbene oligomers but also prenylated flavonoids should be examined further to gain more information on their antimicrobial activity and might be a suitable addition to chemical fungicides on the market to combat gray mold.

High-value compounds obtained from grape canes (Vitis vinifera L.) by steam pressure alkali extraction

The main by-product generated by the wine industry is the grape canes derived from the pruning process. The aim of the current work was the valorisation of this highly polyphenolic lignocellulosic material, by alkali extraction methodology avoiding alcoholic solvents use.In lignocellulosic by-products valorisation, it is important to take into account the variability of the raw material. In the case of vine pruning this may be due to the different varieties used, the geographical area, the climate, or the processing. It is, however, important to develop a global strategy for valorisation, from a biorefinery and circular economy point of view. In this work, the grape canes were characterised, before and after the extraction process, by Fourier transform infrared spectroscopy – attenuated total reflectance (FTIR-ATR) spectroscopy. Changes in cellulose crystallinity, hemicellulose content and structural conformation of proteins, due to the alkali thermal extraction process were identified and evaluated. For the first time, a steam pressure alkali extraction method was used on this material. The products were characterised by FTIR-ATR and Matrix Assisted Laser Desorption Ionisation-Time of Flight (MALDI-TOF) mass spectrometry. The presence of high-value compounds such as stilbene oligomers and condensed tannins was demonstrated in the extracts. The work shows the viability of the steam pressure alkali extraction, a simple and environmentally friendly method, for grape cane valorisation.

Stilbene oligomers from the stems of Parthenocissus tricuspidata and their potential anti-neuroinflammatory and neuroprotective activity

A new stilbene trimer, carasiphenol E (1), together with 13 known stilbenes (2–14) were isolated from the stems of Parthenocissus tricuspidata. The chemical structure of 1 was elucidated by spectroscopic and spectrometric data analyses, and by applying computational methods. All of the isolated compounds (1–14) were assessed for their effect to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated murine microglia BV-2 cells and produce nerve growth factor (NGF) production effect in C6 glioma cells. Compounds 2, 5, 6, and 10–12 inhibited NO production with IC50 values ranging from 14.27 to 37.76 μM. Compounds 4 and 7 induced NGF secretion in C6 glioma cells (132.02 ± 7.55% and 128.15 ± 2.85%, respectively).

Mammalian Arginase Inhibitory Activity of Methanolic Extracts and Isolated Compounds from Cyperus Species

Polyphenolic enriched extracts from two species of Cyperus, Cyperus glomeratus and Cyperus thunbergii, possess mammalian arginase inhibitory capacities, with the percentage inhibition ranging from 80% to 95% at 100 µg/mL and 40% to 64% at 10 µg/mL. Phytochemical investigation of these species led to the isolation and identification of two new natural stilbene oligomers named thunbergin A-B (1–2), together with three other stilbenes, trans-resveratrol (3), trans-scirpusin A (4), trans-cyperusphenol A (6), and two flavonoids, aureusidin (5) and luteolin (7), which were isolated for the first time from C. thunbergii and C. glomeratus. Structures were established on the basis of the spectroscopic data from MS and NMR experiments. The arginase inhibitory activity of compounds 1–7 was evaluated through an in vitro arginase inhibitory assay using purified liver bovine arginase. As a result, five compounds (1, 4–7) showed significant inhibition of arginase, with IC50 values between 17.6 and 60.6 µM, in the range of those of the natural arginase inhibitor piceatannol (12.6 µM). In addition, methanolic extract from Cyperus thunbergii exhibited an endothelium and NO-dependent vasorelaxant effect on thoracic aortic rings from rats and improved endothelial dysfunction in an adjuvant-induced arthritis rat model.

Polyphenolic Characterization of Merlot, Tannat and Syrah Skin Extracts at Different Degrees of Maturity and Anti-Inflammatory Potential in RAW 264.7 Cells.

(1) Background: Both sensory quality and healthy attributes of Vitis vinifera grapes used for winemaking are closely related with the polyphenolic composition of their skins. (2) Methods: In this study, the polyphenolic characterization (flavan-3-ols, procyanidins, flavonols, stilbenes, anthocyanins) was investigated by ultra performance liquid chromatography coupled to a triple quadrupole mass spectrometer (UPLC-QqQ-MS). Skins from Vitis vinifera Merlot, Tannat, and Syrah red grape varieties cultivated in the south of France at different stages of ripening in 2018 were used. The anti-inflammatory and the antioxidant potential of the extracts were evaluated by the measure of nitric oxide (NO) and the intracellular reactive oxygen species production (ROS) in lipopolysaccharide (LPS)-stimulated macrophages. (3) Results: 41 polyphenols were quantified in all samples. Generally, the flavan-3-ol and procyanidin content decreased during ripening whereas the anthocyanins and stilbenes increased. In addition, as a novelty of this work, a wide identification and characterization of monomeric and oligomeric stilbenes was assessed by using authentic standards isolated in our laboratory, some of them (parthenocissin A and miyabenol C) reported for the first time in Merlot, Tannat and Syrah cultivars. The before-veraison skin extracts of all studied varieties, exhibited higher NO and ROS productions inhibition (>50%) proving both antioxidant and anti-inflammatory properties.

Chemical Constituents from Roots of Sophora davidii (Franch.) Skeels and Their Glucose Transporter 4 Translocation Activities.

Sophora davidii (Franch.) Skeels is a multi-purpose traditional medicine that has long been used for the treatment of various diseases. To discover the potential bioactive composition of S. davidii, a chemical investigation was thus performed. In this research, two new stilbene oligomers, Davidiol E–F (1–2), one new 4-aryl-substituted isoflavan Davidinin A (3), and one new 2-arylbenzofuran dimer, Shandougenine C (4), as well as six known compounds (5–10) were obtained from the ethyl acetate fraction of Sophora davidii (Franch.) Skeels. The structures of new compounds were established by extensive 1D and 2D nuclear magnetic resonance (NMR) spectra with mass spectroscopy data. The absolute configuration of 1–3 was assigned by comparing its experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 1–10 promoted glucose transporter 4 (GLUT-4) translocations by the range of 1.28–2.60 folds, respectively. Compound 9 showed the most potent glucose transporter 4 translocations with 1.60 fold enhancement. The result attained in this study indicated that the separation and characterization of these compounds plays an important role in the research and development of new anti-diabetic drugs and pharmaceutical industry.

Characterization and Optimization of the Tyrosinase Inhibitory Activity of Vitis amurensis Root Using LC-Q-TOF-MS Coupled with a Bioassay and Response Surface Methodology.

Vitis amurensis roots have been reported to have the potential for skin whitening through the evaluation of melanogenesis and tyrosinase inhibitory activities. In this study, V. amurensis roots were utilized to quickly select whitening ingredients using LC-Q-TOF-MS coupled with tyrosinase inhibitory assay, and to optimize the extraction process for use as a skin whitening functional material by response surface methodology. Results showed that V. amurensis roots exhibited tyrosinase inhibitory effects by two stilbene oligomers, ε-viniferin (1) and vitisin B (2), as predicted by LC-Q-TOF-MS coupled with bioassay. The optimal extraction conditions (methanol concentration 66%, solvent volume 140 mL, and extraction time 100 min) for skin whitening ingredients were established with the yields 6.20%, and tyrosinase inhibitory activity was 87.27%. The relationship between each factor and its corresponding response was confirmed by Pearson’s correlation analysis. The solvent volume showed clear linear relationship with yields, and methanol concentration had a strong linear relationship with tyrosinase inhibitory activity for compounds 1 and 2, as well as their combination. Overall, LC-Q-TOF-MS coupled with bioassay was proved to have the potential to effectively find new active constituents, as well as known active constituents; vitisin B can be proposed as a new natural potential whitening agent.

Chemical process to improve natural grapevine-cane extract effectivity against powdery mildew and grey mould

Grapevine canes are vine growing byproducts studied for their antimicrobial activities. These properties are directly connected to the stilbene content; oligomeric stilbenes being the most active. In this study, we propose a chemical process, based on oxidative coupling, using metals to increase the oligostilbene rate and the biological effectivity of cane extract against grapevine pathogens. A total of ten compounds were obtained and identified by combining LCMS and NMR spectroscopies, including four newly reported compounds: trans-oxistilbenin C, trans-oxistilbenin D, and cis- and trans-oxistilbenin E. The extract and the main stilbene formed were evaluated for their preventive effects on Plasmopara viticola and Botrytis cinerea growth. The processed extract was highly effective against both pathogens.

Combined MS/MS-NMR Annotation Guided Discovery of Iris lactea var. chinensis Seed as a Source of Viral Neuraminidase Inhibitory Polyphenols.

In this study, the chemical diversity of polyphenols in Iris lactea var. chinensis seeds was identified by combined MS/MS-NMR analysis. Based on the annotated chemical profile, the isolation of stilbene oligomers was conducted, and consequently, stilbene oligomers (1-10) were characterized. Of these, compounds 1 and 2 are previously undescribed stilbene dimer glycoside (1) and tetramer glycoside (2), respectively. Besides, to evaluate this plant seed as a rich source of stilbene oligomers, we quantified three stilbene oligomers of I. lactea var. chinensis seeds. The contents of three major stilbene oligomers—trans-ε-viniferin (3), vitisin A (6), and vitisin B (9)—in I. lactea var. chinensis seeds were quantified as 2.32 (3), 4.95 (6), and 1.64 (9) mg/g dry weight (DW). All the isolated compounds were tested for their inhibitory activities against influenza neuraminidase. Compound 10 was found to be active with the half maximal inhibitory concentration (IC50) values at 4.76 μM. Taken together, it is concluded that I. lactea var. chinensis seed is a valuable source of stilbene oligomers with a human health benefit.

Screening of Natural Stilbene Oligomers from Vitis vinifera for Anticancer Activity on Human Hepatocellular Carcinoma Cells.

The characterization of bioactive resveratrol oligomers extracted from Vitis vinifera canes has been recently reported. Here, we screened six of these compounds (ampelopsin A, trans-ε-viniferin, hopeaphenol, isohopeaphenol, R2-viniferin, and R-viniferin) for their cytotoxic activity to human hepatocellular carcinoma (HCC) cell lines p53 wild-type HepG2 and p53-null Hep3B. The cytotoxic efficacy depended on the cell line. R2-viniferin was the most toxic stilbene in HepG2, with inhibitory concentration 50 (IC50) of 9.7 ± 0.4 µM at 72 h, 3-fold lower than for resveratrol, while Hep3B was less sensitive (IC50 of 47.8 ± 2.8 µM). By contrast, hopeaphenol (IC50 of 13.1 ± 4.1 µM) and isohopeaphenol (IC50 of 26.0 ± 3.0 µM) were more toxic to Hep3B. Due to these results, and because it did not exert a large cytotoxicity in HH4 non-transformed hepatocytes, R2-viniferin was selected to investigate its mechanism of action in HepG2. The stilbene tended to arrest cell cycle at G2/M, and it also increased intracellular reactive oxygen species (ROS), caspase 3 activity, and the ratio of Bax/Bcl-2 proteins, indicative of apoptosis. The distinctive toxicity of R2-viniferin on HepG2 encourages research into the underlying mechanism to develop the oligostilbene as a therapeutic agent against HCC with a particular genetic background.

Structural Studies on Stilbene Oligomers Isolated from the Seeds of Melinjo (Gnetum gnemon L.).

This paper describes the isolation and structural determination of a new stilbene dimer, named 7a-epi-gnetin C, from melinjo (Gnetum gnemon L.) seed extract. The relative structure was elucidated based on NMR spectroscopic evidence, while the absolute configuration was assigned by a combination of NMR and electronic circular dichroism spectroscopic analysis and chemical conversion. 7a-epi-Gnetin C was evaluated as an antioxidant and was shown to have a comparable activity to the known stilbene oligomers. In addition, the structural revision of gnetin L, a known stilbene dimer, was also discussed.

A dimeric stilbene extract produced by oxidative coupling of resveratrol active against <em>Plasmopara viticola</em> and <em>Botrytis cinerea</em> for vine treatments

Aim: Stilbenes are a group of polyphenols that contribute to the defense mechanisms of grapevine under stress conditions. The main active compounds are oligomeric stilbenes. The sourcing of such compounds is limited by plant raw material availability. In order to overcome this limitation, we proceeded to hemisynthesis of oligomeric stilbenes from natural resveratrol by oxidative coupling using metals. The reaction mixture and the obtained pure compounds were tested for their antimicrobial activity in vitro against P. viticola and B. cinerea, and compared with that of resveratrol and grapevine cane extract.Methods and results: Hemisynthesis was achieved in methanol using silver acetate (AgOAc) as oxidative coupling reagent. Four main compounds were obtained in the reaction mixture: δ-viniferin, parthenostilbenin B, oxistilbenins A and B. Results demonstrate the ability of oxidative coupling reaction to produce potent inhibitors of P. viticola and B. cinerea. The reaction mixture is enriched in antimicrobial compounds with high potency as it is at least eight times more active than grapevine cane extract against P. viticola. In addition, this reaction mixture reveals a high fungitoxicity against B. cinerea in contrast to the grapevine extract.Conclusion: The present findings indicate that metals could be used for producing highly antimicrobial compounds from natural resveratrol.Significance and impact of the study: In this study, a method to produce active dimeric stilbenes to protect grapevine was developed based on botanical ingredients. The combination of natural extracts, rich in resveratrol, and metals such as copper may be used as alternatives to Bordeaux mixture and chemical fungicides to protect crops.

Resveratrol transformation in red wine after heat treatment

Resveratrol is a well-known wine constituent. Its concentration can vary according to the cultivar choice and the winemaking process. Due to its phenolic structure, resveratrol could be transformed under high temperature or oxidative conditions, leading to the formation of various derivatives including oligomers. Hence, the goal of this study is to investigate the presence of these derivatives in wine. In the first stage, hemisynthesis of oligomeric stilbenes was achieved from resveratrol in ethanol by oxidative coupling using metals. Four de novo synthetized resveratrol derivatives were identified by MS and NMR spectroscopy including two new molecules, oxistilbenin F and oxistilbenin G. In the second stage, analysis of red wine after heat treatment by LC-MS confirmed the presence of some of these compounds in wine. Finally, the anti-inflammatory effects of the compounds were evaluated by studying their ability to prevent lipopolysaccharide (LPS)-induced upregulation of nitric oxide (NO) and reactive oxygen species (ROS) production in RAW 264.7 macrophage cell line.

Subcritical water extraction of stilbenes from grapevine by-products: A new green chemistry approach

Currently, one of the biggest challenges in viticulture is to valorize the huge amount of biomass, especially grapevine waste. Subcritical water is a sustainable and eco-friendly method of extraction based on the use of water as an extracting agent, corresponding to the strategy called "green chemistry". Application of a high temperature and a high pressure maintains the water in its liquid form allowing polar and less polar compounds to be extracted. Stilbenes are low-polarity compounds that normally require organic solvents for their extraction. This study investigated the possibility of using a subcritical water process to extract complex stilbenes from three vine co-products: cane, wood, and root. Various temperatures (100, 130, 160 and 190 °C) and contact times (5, 15 and 30 min) were tested by accelerated solvent extraction (ASE). The highest stilbene yield was obtained at 160 °C and 5 min for cane (3.62 g/kg dry mass), wood (9.32 g/kg dry mass) and root (12.1 g/kg dry mass). ASE allowed the extraction of monomers, dimers, trimers and tetramers of stilbenes in similar amounts as with conventional organic solvents. In addition, the subcritical extraction conditions led to the formation of an uncommon tetrameric stilbene, named vitisin F. The possibility of using “green chemistry” for extracting oligomeric stilbenes is a step forward in the search for alternatives to organic solvents.

ANTIBACTERIAL AND CYTOTOXIC POTENCIES OF STILBENE OLIGOMERS FROM STEM BARKS OF BAOTI (DRYOBALANOPS LANCEOLATA) GROWING IN KENDARI, INDONESIA

Objective: The purpose of the research is to isolate and to identify compounds from stem barks of Dryobalanops lanceolata and to evaluate their activities toward bacteria and cancer cell lines.Methods: Isolation of the compounds worked with chromatography methods including thin-layer chromatography, vacuum liquid chromatography, and radial chromatography. All compound structures were determined based on the spectroscopic evidence including ultraviolet (UV), Infrared, and one-dimension and two-dimension nuclear magnetic resonance spectra and comparison the spectroscopy data with related data from references. The biological properties of compounds are evaluated toward bacteria Escherichia coli ATCC 35219 and Staphylococcus aureus ATCC 25923 and human breast cancer cell lines T-47D as cytotoxic potency.Results: Five stilbene oligomers have been isolated and identified from acetone extract of D. lanceolata stem barks namely balanocarpol (1), e-viniferin (2), a-viniferin (3), vaticanol B (4), and hopeaphenol (5). The inhibition zone value (in mm) of e-viniferin, balanocarpol, a-viniferin, hopeaphenol, and vaticanol B toward E. coli and S. aureus is 11±0.22 and 7±0.17, 9±0.17 and 13±0.12, 8±0.20 and 8±0.16, 6±0.16 and 8±0.11, and 5±0.12 and 4±0.14, respectively. Biological activity of the compounds against breast cancer cell lines T-47D (inhibitory concentration 50% in μM) for e-viniferin, balanocarpol, a-viniferin, hopeaphenol, and vaticanol B is 34.13±0.15, 98.17±0.41, 84.79±0.24, 52.04±0.26, and 119.30±0.54, respectively.Conclusions: e-viniferin is the most active compound toward E. coli and human breast cancer cell lines T-47D. Biological activity against bacteria S. aureus indicated that balanocarpol is the most potential compound.

(–)-Ampelopsin F, Dimerstilbene Compound from Dryobalanops oblongifolia and Antimalarial Activity Test

(–)-Ampelopsin F, Dimerstilbene Compound from Dryobalanops oblongifolia and Antimalarial Activity Test

Two new stilbene trimers from Cynodon dactylon

Many naturally occurring oligostilbenes have drawn considerable attention because of their intricate structures and diverse bioactivities. Two new stilbene trimers, cystibenetrimerol A (1) and cystibenetrimerol B (2) were isolated from the dried grass of Cynodon dactylon (L.) Pers. The planar structures and stereo configurations of them were elucidated by spectroscopic and spectrometric methods. The isolation and structures elucidation of two new stilbene trimers suggested the ordinary grass belonging to the family Poaceae may be a rich source of stilbene oligomers.