Phenethyl Alcohol Glycosides Research Articles

UPLC-Q-TOF-MS based investigation into the bioactive compounds and molecular mechanisms of Lamiophlomis Herba against hepatic fibrosis

BackgroundLamiophlomis Herba (LH) is a valuable traditional medicinal plant found on the Qinghai-Tibetan Plateau that promotes blood circulation, removes blood stasis, and has antibacterial and anti-inflammatory properties. The main components of LH are iridoid glycosides, phenethyl alcohol glycosides, flavonoids, and polysaccharides. PurposeTo investigate the mechanism of the anti-liver fibrosis effects of LH and screen for its bioactive compounds. Study designScreening LH marker components and validating the LH anti-liver fibrosis mechanism. MethodsThe active ingredients of LH were identified using UPLC-Q-TOF-MS, and HotMap combined with principal components analysis (PCA) was used to screen for marker components. Network pharmacology and molecular docking techniques were used to predict the potential anti-fibrotic targets of LH. Immunofluorescence, enzyme-linked immunosorbent assay (ELISA), real-time PCR (RT-PCR), and western blotting were used for experimental validation and mechanistic studies. ResultsFifteen compounds that actively contributed to the cluster were identified as marker compounds. Acteoside, 8-O-acetyl shanzhiside methyl ester (8-O-ASME), Luteolin, Shanzhiside Methyl ester (SME), Loganin, Loganate were the main active components. Network pharmacology and molecular docking studies have shown that LH might improve liver fibrosis, inflammation, and oxidative stress, which might be related to key targets such as PTGS2, MAPK, EGFR, AKT1, SRC, Fn1, Col3a1, Col1a1, and PC-III. The results of ELISA, RT-PCR and western blot experiments showed that Acteoside, 8-O-ASME, Luteolin, SME, Loganin, Loganate, and the LH group could reduce the levels of fibronectin, Col1a1, Col3a1, α-SMA, Col-Ⅳ, LN, and PC-Ⅲ. ConclusionLH improves liver fibrosis induced by HSC-T6 cells and inhibits the deposition of extracellular matrix (ECM) in hepatocytes, resulting in a decrease in the degree of liver fibrosis and a good anti-liver fibrosis effect.

A previously undescribed phenylethanoid glycoside from Callicarpa kwangtungensis Chun acts as an agonist of the Na/K-ATPase signal transduction pathway

The new concept that Na/K-ATPase acts as a receptor prompted us to look for new ligands from Callicarpa kwangtungensis Chun. Using column chromatography, an undescribed phenethyl alcohol glycoside, callicarpanoside A, and an undescribed benzyl alcohol glycoside, callicarpanoside B, along with twelve known polyphenols were isolated from Callicarpa kwangtungensis Chun. All the isolated compounds were evaluated for their Na/K-ATPase (NKA) inhibitory activities. Using our NKA technology platform-based screening assay protocols, callicarpanoside B was identified as an undescribed Na/K-ATPase agonist. In particular, the newly identified benzyl alcohol glycoside was found to bind NKA and activate the receptor NKA/Src complex, resulting in the activation of protein kinase cascades. These cascades included extracellular signal-regulated kinases and protein kinase C epsilon, as well as NKA α1 endocytosis at nanomolar concentrations. Unlike the class of cardiotonic steroids, callicarpanoside B showed less inhibition of NKA activity and caused less cellular toxicity. Moreover, callicarpanoside B was found to bind NKA at a different site other than the cardiotonic steroids binding site. Thus, we have identified an undescribed NKA α1 agonist that may be used to enhance the physiological processes of NKA α1 signaling.

Bioactive constituents of Lindernia crustacea and its anti-EBV effect via Rta expression inhibition in the viral lytic cycle.

Lindernia crustacea (L.) F.Muell. (Scrophulariaceae) was selected for phytochemical investigation owing to its traditional use against human herpes virus infection and its anti-Epstein-Barr virus (EBV) effect. The present study focused on the phytochemical investigation of L. crustacea including the isolation and structure determination of its biologically active compounds. Compounds with anti-EBV effects were also investigated. The EtOH extract of L. crustacea was subsequently partitioned using different solvents. The EtOAc fraction was subjected to several chromatographic methods to obtain pure compounds. The structures of all isolates were established by spectroscopic analysis and compared with previously reported physical data. The anti-EBV effect was evaluated in an EBV-containing Burkitt's lymphoma cell line (P3HR1) to study the expression of EBV lytic proteins. Thirty-three compounds, including one diterpene (1), four anthraquinones (2-5), two ionones (6 and 7), fourteen phenylpropanoid glycosides (8-21), five flavonoids (22-26), one lignan glycoside (27), one phenethyl alcohol glycoside (28), one phenylpropene glycoside (29), one glucosyl glycerol derivative (30), one furanone (31), and two cinnamic acid derivatives (32 and 33), were isolated from the ethanolic extract of the plant. All isolated compounds were obtained for the first time from Lindernia sp. The evaluation of the anti-EBV activity of L. crustacea crude extract, partitioned fractions, and constituents was performed for the first time. Phytol (1), aloe-emodin (2), byzantionoside B (7), a mixture of trans-martynoside (8) and cis-martynoside (9), a mixture of trans-isomartynoside (10) and cis-isomartynoside (11), luteolin-7-O-β-D-glucopyranoside (24), and apigenin-7-O-[β-D-apiofuranosyl (1→6)-β-D-glucopyranoside] (25) exhibited significant inhibitory effects on the EBV lytic cycle at 20μg/mL in the immunoblot analysis. On the other hand, (6R,7E,9R)-3-oxo-α-ionol-β-D-glucopyranoside (6) and a mixture of trans-dolichandroside A (12) and cis-dolichandroside A (13) showed moderate anti-EBV activity at 20μg/mL. L. crustacea and its active isolates could be developed as potential candidates against EBV. Our findings provide scientific evidence for the traditional use of L. crustacea for its antiviral effects.

Chemomics-based marker compounds mining and mimetic processing for exploring chemical mechanisms in traditional processing of herbal medicines, a continuous study on Rehmanniae Radix

Exploring processing chemistry, in particular the chemical transformation mechanisms involved, is a key step to elucidate the scientific basis in traditional processing of herbal medicines. Previously, taking Rehmanniae Radix (RR) as a case study, the holistic chemome (secondary metabolome and glycome) difference between raw and processed RR was revealed by integrating hyphenated chromatographic techniques-based targeted glycomics and untargeted metabolomics. Nevertheless, the complex chemical transformation mechanisms underpinning the holistic chemome variation in RR processing remain to be extensively clarified. As a continuous study, here a novel strategy by combining chemomics-based marker compounds mining and mimetic processing is proposed for further exploring the chemical mechanisms involved in herbal processing. First, the differential marker compounds between raw and processed herbs were rapidly discovered by untargeted chemomics-based mining approach through multivariate statistical analysis of the chemome data obtained by integrated metabolomics and glycomics analysis. Second, the marker compounds were mimetically processed under the simulated physicochemical conditions as in the herb processing, and the final reaction products were chemically characterized by targeted chemomics-based mining approach. Third, the main chemical transformation mechanisms involved were clarified by linking up the original marker compounds and their mimetic processing products. Using this strategy, a set of differential marker compounds including saccharides, glycosides and furfurals in raw and processed RR was rapidly found, and the major chemical mechanisms involved in RR processing were elucidated as stepwise transformations of saccharides (polysaccharides, oligosaccharides and monosaccharides) and glycosides (iridoid glycosides and phenethylalcohol glycosides) into furfurals (glycosylated/non-glycosylated hydroxymethylfurfurals) by deglycosylation and/or dehydration. The research deliverables indicated that the proposed strategy could advance the understanding of RR processing chemistry, and therefore may be considered a promising approach for delving into the scientific basis in traditional processing of herbal medicines.

Forsythoside A exerts an anti-endotoxin effect by blocking the LPS/TLR4 signaling pathway and inhibiting Tregs in vitro.

Endotoxins, also referred to as lipopolysaccharides(LPS), are powerful immunostimulators involved in a number of severe diseases. ForsythosideA (FTA), a monomer of phenethyl alcohol glycosides extracted from Forsythia suspensa, has been shown to possess anti-bacterial and immunomodulatory properties. However, it is currently not known whether FTA can counter the adverse effects of endotoxins. We investigated the effect of FTA on LPS-stimulated RAW264.7 cells and primary lymphocytes to determine its molecular mechanism of action. RAW264.7 cells and primary lymphocytes were incubated with or without LPS (100ng/ml) in the presence or absence of FTA or polymyxinB. We found that FTA increased the viability of LPS-treated RAW264.7 cells and primary lymphocytes suggesting that FTA effectively counters the adverse effects of endotoxins. FTA decreased the percentage of regulatory Tcells (Tregs) and inhibited the TLR4/MyD88/NF-κB signaling pathway, downregulating Foxp3, IL-10 and TGF-β1, molecules involved in the immunosuppressive function of Tregs. These findings elucidate the molecular mechanism underlying the anti-endotoxin effects of FTA and suggest its use as a new treatment for LPS-induced diseases.

A new phenethyl alcohol glycoside from Orobanche coerulescens

The constituents of the whole plant of Orobanche coerulescens were isolated and purified by using various column chromatographic techniques including D101, silica gel and ODS. The structures were identified by spectroscopic analyses including NMR and MS. A new phenylethanol glycoside was isolated from the whole plant of O. coerulescens, and was identified as 2-(3-methoxy-4-hydroxyphenyl)-ethanol-1-O- [(1→3)-O-α-L-rhamnopyranosyl-4, 6-O-di-feruloyl]-β-D-glucopyranoside, named as orobancheoside B. Through the antibacterial activity test, orobancheoside B was proved to have certain antibacterial activity, and be one of the main active components of O. coerulescens. The research result will laid a foundation for the medicinal materials and quality control research. Activity screening, broomrape orobancheoside B has certain antibacterial activity, as one of the main active components of O. coerulescens, and to constantly improve the quality of the medicinal materials laid a foundation.

Ultra-high performance liquid chromatography coupled with photo-diode array and quadrupole/time-of-flight mass spectrometry based chemical profiling approach to evaluate the influence of preparation methods on the holistic quality of Qiong-Yu-Gao, a traditional complex herbal medicine

Qiong-Yu-Gao (QYG), consisting of Rehmanniae Radix (RR), Poriae (PO) and Ginseng Radix (GR), is a commonly used tonic traditional complex herbal medicine (CHM). So far, three different methods have been documented for preparation of QYG, i.e. method 1 (M1): mixing powders of GR and PO with decoction of RR; method 2 (M2): combining the decoction of RR and PO with the decoction of GR; method 3 (M3): decocting the mixture of RR, GR and PO. In present study, an ultra-high performance liquid chromatography coupled with photo-diode array and quadrupole/time-of-flight mass spectrometry (UHPLC-PDA-QTOF–MS/MS) based chemical profiling approach was developed to investigate the influence of the three preparation methods on the holistic quality of QYG. All detected peaks were unambiguously identified by comparing UV spectra, accurate mass data/characteristic mass fragments and retention times with those of reference compounds, and/or tentatively assigned by matching empirical molecular formula with that of known compounds, and/or elucidating quasi-molecular ions and fragment ions referring to information available in literature. A total of 103 components, mainly belonging to ginsenosides, phenethylalcohol glycosides, iridoid glycosides and triterpenoid acids, were identified, of which 5 degraded ginsenosides were putatively determined to be newly generated during preparation procedures of QYG samples. Triterpenoid acids and malonyl-ginsenosides were detected only in M1 samples, while degraded ginsenosides were merely detectable in M2/M3 samples. The possible reasons for the difference among chemical profiles of QYG samples prepared with three methods were also discussed. It could be concluded that preparation method do significantly affect the holistic quality of QYG. The influence of the altered chemical profiles on the bioactivity of QYG needs further investigation. The present study demonstrated that UHPLC-PDA-QTOF–MS/MS based chemical profiling approach is efficient and reliable for evaluating the holistic quality of traditional CHM.

Rapid Identification of Chemical Constituents in Traditional Chinese Medicine Fufang Preparation Xianling Gubao Capsule by LC-linear Ion Trap/Orbitrap Mass Spectrometry

The traditional Chinese medicine fufang preparation "Xian-Ling-Gu-Bao" capsule (XLGB), which is composed of six herbal medicines, is popularly used for the treatment of osteoporosis. A reliable and effective method using LC-linear ion trap (LTQ)/Orbitrap mass spectrometry for rapid screening and identification of chemical constituents in "Xian-Ling-Gu-Bao" capsule is described in this paper. Based on the UV spectrum, mass spectrum, and the chemical components isolated from the original plants of XLGB, 118 compounds were identified or tentatively characterized, including 58 flavonoid glycosides, six prenylated flavonones, five prenylated isoflavones, six prenylated chalcones, four xanthone C-glycosides, 13 saponins, eight phenolic acids, five coumarins, three lignans, three iridoids, five phenethyl alcohol glycosides, one tanshinone and one alkaloid. This work might be helpful for the quality control and further pharmacokinetic studies of XLGB, and provided a good example for the rapid identification of chemical constituents in traditional Chinese medicine fufang preparation. Moreover, the identification strategy for the linkages of sugar residues in flavonol O-glycosides was summarized in the study. The diagnostic fragment ions at m/z 185 [C12H9O2] and 157 [C11H9O], which distinguish C-6 and C-8 prenylated flavonoids, were reported for the first time.

Simultaneous determination of iridoid glycosides, phenethylalcohol glycosides and furfural derivatives in Rehmanniae Radix by high performance liquid chromatography coupled with triple-quadrupole mass spectrometry

In this study, a sensitive and selective method for simultaneously quantifying eight major components (four iridoid glycosides, three phenethylalcohol glycosides and one furfural derivative) of Rehmanniae Radix by high performance liquid chromatography coupled with triple-quadrupole mass spectrometry (HPLC–TQ-MS) was developed. The sample preparation was executed using an optimised ultrasonic method with complete extraction efficiencies of eight analytes. For mass spectrometry, selected ion recording (SIR) scan mode was used to improve the sensitivity and selectivity. The established method was validated in terms of linearity, sensitivity, precision, accuracy and stability, and successfully applied to determine the contents of the eight analytes in different batches of raw and processed Rehmanniae Radix, which confirmed that the established method was reliable and useful for “holistic” quality evaluation of Rehmanniae Radix. The quantitative results indicated that the quality of commercial raw or processed Rehmanniae Radix was remarkably inconsistent.

Bumaldosides A, B and C from the Leaves of Staphylea bumalda

Two new aliphatic diglycosides and a phenolic glucoside (4, 5 and 7) have been isolated from leaves of Staphylea bumalda DC., together with three known compounds, benzyl and phenethyl alcohol glycosides (1 and 2), and zingerone β-D-glucopyranoside (6). 2-Ethyl-3-methylmaleimide N-glucopyranoside (3) was first isolated as a free form. Their structures were determined on the basis of spectroscopic analysis.

New phenethyl alcohol glycosides from Stachys parviflora

Phytochemical investigations of the whole plant of Stachys parviflora (Lamiaceae) resulted in the isolation of two new phenethyl alcohol glycosides. The structures of the new compounds named parviflorosides A and B were established as 2-(3,4-dihydroxyphenyl)-ethyl-O-α-l-rhamnopyranosyl-(1 → 2)-4-O-E-caffeoyl-β-d-glucopyranoside (1) and 2-(3,4-dihydroxyphenyl)-ethyl-O-α-l-rhamnopyranosyl-(1 → 2)-6-O-E-caffeoyl-β-d-glucopyranoside (2), respectively. The structure elucidation of the new compounds was based primarily on 1D and 2D NMR analysis, including COSY, HMBC and HMQC correlations.

Remophilanetriol: A New Eremophilane from the Roots of Rehmannia glutinosa

AbstractFor Abstract see ChemInform Abstract in Full Text.

Remophilanetriol: A New Eremophilane from the Roots of Rehmannia glutinosa

Radix Rehmanniae is the fresh or dry root of Rehmannia glutinosa Libosch. (Scrophulariiaceae), and is traditionally used for antipyretic and hemostatic. The roots of Rehmannia glutinosa have been also used as a tonic and sedative after processing of the fresh root such as boiling in the rice wine or steaming. A number of constituents such as iridoids, phenethyl alcohol glycosides, glycosides, cyclopentanoid monoterpenes, and norcarotenoids, have been reported from the fresh or processed roots of R. glutinosa. In the course of chemical investigation on the dried roots of R. glutinosa, a new eremophilane-type metabolite, named remophilanetriol (1), was encountered, together with aeginetic acid (2), which has been identified from the same species. Details of the isolation and structure elucidation of new compound (1) are described in the present study. The MeOH extract of Rehmannia glutinosa was concentrated, suspended in H2O, and sequentially partitioned with n-hexane, EtOAc, and BuOH. Purification of the EtOACsoluble extract by utilizing normal-phase and reversed-phase column chromatography, followed by reversed-phase HPLC, resulted in the isolation of compounds 1 and 2. Compound 1 has the molecular formula of C15H24O4, as deduced from the C NMR and HRFABMS data, which indicated four degrees of unsaturation. Analysis of H, C and DEPT NMR data (Table 1) revealed the presence two methyls, six methylenes (two oxygenated), three methines, and four quaternary carbon including carbonyl (δ 199.8) functionality. These data accounted 21 carbon-bound protons, requiring the presence of three hydroxyl groups in the molecule. In addition, carbon resonances observed at δ 199.8, δ 161.6, and δ 124.4 were strongly suggestive of the presence of an α,β-unsaturated ketone moiety. Since the above structure information accounted for all but two unsaturation equivalents, compound 1 must be bicyclic to satisfy four degrees of unsaturation required by its molecular formula. The structure of remophilanetriol was unambiguously deduced by analysis of 2D NMR data (COSY, HMQC, and HMBC). Analysis of COSY results led to the

Megastigmane, benzyl and phenethyl alcohol glycosides, and 4,4'-dimethoxy-beta-truxinic acid catalpol diester from the leaves of Premna subscandens MERR.

Extensive isolation work on the n-BuOH-soluble fraction obtained from the leaves of Premna subscandens, collected on Ishigaki island, Okinawa, afforded six compounds. Two were identified as megastigmane glucosides, 7-(3,5-dihydroxy-1,1,5-trimethylcyclohexylidene)-9-methylprop++ +-8-enyl 9-O-beta-D-glucopyranoside and 3-hydroxy-5,6-epoxy-beta-ionol 9-O-beta-D-glucopyranoside. The structures of the remaining four new compounds were elucidated to be a 2'-O-beta-D-apiofuranosyl derivative of 3-hydroxy-5,6-epoxy-beta-ionol 9-O-beta-D-glucopyranoside, named premnaionoside, benzyl alcohol beta-D-(2'-O-beta-D-xylopyranosyl)glucopyranoside, phenethyl alcohol beta-D-(2'-O-beta-D-glucopyranosyl)glucopyranoside, and 4,4'-dimethoxy-beta-truxinic acid catalpol diester by spectroscopic analyses.

Phenethyl alcohol glycosides and isopentenol glycoside from fruit of Bupleurum falcatum.

Investigation on the constituents of the fruit of Bupleurum falcatum L. resulted in the isolation of the three new glycosides, phenethyl alcohol 8-O-beta-D-glucopyranosyl-(1-->2)-O-beta-D-apiofuranosyl-(1-->6)-b eta-D- glucopyranoside, phenethyl alcohol 8-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside and isopentenol 1-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside along with five known glycosides, icariside D1, icariside F2, saikosaponin a, saikosaponin c and saikosaponin d. The structures of these compounds were elucidated on the basis of interpretation of chemical and spectral data.

New Glycosides from Ajuga decumbens

A new phenethyl alcohol glycoside, galactosylmartynoside (1), and a new abietatriene-type diterpene glycoside, ajugaside A (2), were isolated from the whole plants of Ajuga decumbens, together with known phenethyl alcohol glycosides (3 and 4) and iridoid glycosides (5-7). Chemical structures were elucidated on the basis of spectral data. Of these compounds, 8-acetylharpagide (6) exhibited the strongest inhibitory effect on Epstein-Barr virus activation induced by 12-O-tetradecanoylphorbol-13-acetate.

Two phenethyl alcohol glycosides from Scutellaria orientalis subsp. Pinnatifida

Two new phenethyl alcohol glycosides, darendoside A and B (= deacyl martynoside) were isolated from the methanolic extract of the aerial parts of Scutellaria orientalis subsp. pinnatifida, along with four known glycosides, syringin, martynoside, leucosceptoside A and verbascoside. On the basis of chemical and spectral evidence the structures of darendoside A and B were determined as β-(4-hydroxyphenyl) ethyl O-β- d-apiofuranosyl-(1→ 2)- O-β- d-glucopyranoside and β-(3-hydroxy-4-methoxyphenyl)ethyl O-α- l-rhamnopyranosyl- (1→3)- O-β- d-glucopyranoside (=deacyl martynoside), respectively.

Premnaodorosides A, B and C, iridoid glucoside diesters of an acyclic monoterpenediol from leaves of Premna odorata

The acyclic monoterpenediol diesters, premnaodorosides A, B and C, isolated from Premna odorata, were shown to be 3,7-dimethyloctan-1,8-diol esterified with iridoid acid glucosides, namely two mussaenosidic acid moieties, one mussaenosidic moiety and one 8-epiloganic acid moiety, and one mussaenosidic acid moiety and one gardoside moiety. Phenethyl alcohol glycosides, verbascoside, isoacteoside, bioside (decaffeoylverbascoside), and cistanoside F were also isolated.

Nine phenethyl alcohol glycosides from Stachys sieboldii

Three new phenethyl alcohol glycosides together with six known compounds have been isolated from the leaves of Stachys sieboldii. On the basis of chemical and spectral analyses, the structures of three new compounds named stachysosides A, B and C have been established as 2-(3,4-dihydroxyphenyl)ethyl O-α- l-arabinopyranosyl-(1→2)-α- l-rhamnopyranosyl-(1→3)-4- O- E-caffeoyl-β- d-glucopyranoside, 2-(3,4-dihydroxyphenyl)ethyl O-α- l-arabinopyranosyl-(1→2)-α- l-rhamnopyranosyl-(1→3)-4- O- E-feruloyl-β- d-glucopyranoside and 2-(3-hydroxy-4-methoxyphenyl)ethyl O-α- l-arabinopyranosyl-(1→2)-α- l-rhamnopyranosyl-(1→3)-4- O- E-feruloyl-β- d-glucopyranoside, respectively.

Iridoid diglycoside monoacyl esters from stems of Premna japonica

From a methanol extract of stems of Premna japonica, three new monoacyl 6- O-α- l-rhamnopyranosylcatalpols were isolated. They were determined to be the 2 - ̋ O-,3″- O- and 4″- O-feruloyl esters of 6- O-α- l-rhamnopyranosycatalpol. Along with these compounds, two phenethyl alcohol glycosides, acteoside and martynoside, aucubin and 6- O-α- l-(2″- O- p- methoxycinnamoyl)rhamnopyranosylcatalpol were also isolated.