Two new cadinane-type sesquiterpenoid glycosides from Dryopteris fragrans with anti-inflammatory activities.

Two new cadinane-type sesquiterpenoid glycosides, dryopteristerpeneA (1) and dryopteristerpeneB (2), were isolated from the aqueous extract of Dryopterisfragrans. Their structures were determined by spectroscopic data analysis and electronic circular dichroism (ECD) calculations. Compounds 1 and 2 exhibited inhibition on nitric oxide production in lipopolysaccharide induced RAW 264.7 macrophages with their IC50 values of 60.5 and 59.8 µM, respectively.

[A new phloroglucinol compound from Dryopteris fragrans].

Two phloroglucinol compounds(1-2) were isolated and purified from 95% ethanol extract of Dryopteris fragrans through various column chromatographies on silica gel, Sephadex LH-20, medium pressure column chromatography, and preparative HPLC. Their structures were elucidated as 2',4',6'-trihydroxy-5'-methyl acetate-3'-methyl-1'-butyrophenone(1) and aspidinol B(2) based on their chemical and physicochemical methods and spectroscopic data. Compound 1 is a new phloroglucinol compound named "dryofraginol".

Anti-Influenza Virus (H5N1) Activity Screening on the Phloroglucinols from Rhizomes of Dryopteris crassirhizoma.

For screening the active phloroglucinols on influenza virus (H5N1) from Dryopteris crassirhizoma NaKai, a database was established including twenty-three phloroglucinols that had been isolated from Dryopteris crassirhizoma. Their inhibitory effect on the neuraminidase (NA) of influenza virus H5N1 was screened by molecular docking. As a result, three candidates were selected. The rhizomes of D. crassirhizoma were subjected to isolation and purification processes to obtain the inhibitor candidates. Thirteen phloroglucinols were obtained, including three selected candidates and two new phloroglucinols. The five phloroglucinols were investigated for their inhibitory activity on NA in vitro. The results showed that dryocrassin ABBA and filixic acid ABA exhibited inhibitory effects on NA with IC50 as 18.59 ± 4.53 and 29.57 ± 2.48 µM, respectively, and the other three phloroglucinols showed moderate inhibitory activity. Moreover, the anti-influenza virus (H5N1) activity and cytotoxicity of dryocrassin ABBA and filixic acid ABA were tested on Madin-Darby canine kidney (MDCK) cells with the cell counting kit-8 (CCK8) method. The results confirmed that dryocrassin ABBA exhibited an inhibitory activity with low cytotoxicity (TC50 > 400 µM) against influenza virus (H5N1) which will have to be investigated in further detail. In conclusion, phloroglucinols from D. crassirhizoma were shown to have anti-influenza virus activity, and especially dryocrassin ABBA, one of the phloroglucinols, may have the potential to control influenza virus (H5N1) infection.

Two new glycosides from Dryopteris fragrans with anti-inflammatory activities.

Phytochemical investigation on the aqueous extract from Dryopteris fragrans led to the isolation of one new chromone glycoside, frachromone C (1), and one new coumarin glycoside, dryofracoulin A (2), together with one known undulatoside A (3). Their structures were elucidated by a combination of 1D and 2D NMR, HRMS, and chemical analysis. Compounds 1-3 exhibited inhibition on nitric oxide production in lipopolysaccharide induced RAW 264.7 macrophages with their IC50 values of 45.8, 65.8, and 49.8 µM, respectively.

[Analysis of digital gene expression profiles of the cultivated Polygala tenuifolia in different phenological phases].

The content changes of chemical components in different phenological phase of the cultivated Polygala tenuifolia is one of the important factors for determination of the best harvest time in the production practice. In this study, the digital gene expression (DGE) profiles of the cultivated P. tenuifolia were analyzed in different phenological phase (flowering fruit bearing stage, wilting stage, dormancy stage). The differentially expressed genes were found in the biosynthesis of chemical composition in P. tenuifolia, and the representational ones were validated by RT-q PCR. Then, the key enzymes(CYP450s and UGTs) involved in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia were predicted through the correlation analysis of gene expression. The number of down-regulated genes was more than that of up-regulated in P. tenuifolia from flowering fruit bearing stage to dormancy stage. Six differentially expressed genes (HMGS, PMK, FPPS, SQS, SE, ß-AS) and five (PAL, C4 H, 4CL, CAD, peroxidase) were annotated to the triterpenoid saponins and phenylpropanoid biosynthesis pathway in P. tenuifolia, respectively. Compared to wilting and dormancy stages, the saponins, xanthones, and lignins were largely synthesized at the flowering fruit bearing stage of P. tenuifolia. Furthermore, UGT83A1, CYP716B1, CYP98A3, CYP86B1, and CYP94A1 may be the part of key enzymes in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia. This study provides evidence to support the correctness of traditional harvest time of P. tenuifolia at the level of transcription, and lays the scientific foundation for gene cloning and functional verification of CYP450 s and UGTs in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia in the future.

[UPLC/Q-TOF MS and NMR plant metabolomics approach in studying the effect of growth year on the quality of Polygala tenuifolia].

Growth year is one of the important factors for the quality of Polygala tenufolia. In this study, primary metabolites and secondary metabolites were compared in 1, 2 and 3 years old P. tenufolia cultivated in Shaanxi Heyang. The samples were subjected to ultra-high performance liquid chromatography (UPLC)-quadrupole time-of-flight mass spectrometry (Q-TOF MS) and nuclear magnetic resonance (NMR) analysis, and the obtained data were analyzed using principal component analysis (PCA) and other statistical analysis methods. In addition, content and correlation of different metabolites were also calculated. The results showed no significance between main component contents in 2 year-old and 3 year-old P. Tenufolia, but 1 year-old was statistically different. The contents of primary metabolites, such as fructose, sucrose, and choline increased as time goes on, while glycine and raffinose decreased. The contents of secondary metabolites, such as onjisaponin Fg, polygalasaponin XXVIII, polygalasaponin XXXII increased, while polygalaxanthone III and parts of oligosaccharide multi-ester including tenuifoliose A, tenuifoliose C, tenuifoliose C2 and tenuifoliose H decreased with the extension of the growth years. Growth years has important impact on the quality of P. tenuifolia and the existing growing years of commodity P. tenuifolia have its scientific evidence. This study supplied a new method for the quality evaluation of Chinese medicinal materials.

[UPLC/Q-TOF MS-Based Metabolomics Analysis of Chemical uiterences in Different Polygala tenuifolia Varieties].

OBJECTIVE: The chemical differences of Polygala tenuifolia varieties-JinYuan 1 (JY1), FenYuan 2 (FY2) and traditional FenYang (FY) were studied, in order to provide reference for the breeding of Polygala tenuifolia. METHODS: The samples of JY1, FY2 and FY were subjected to ultra-high performance liquid chromatography (UPLC) quadrupole time-of-flight mass spectrometry (Q-TOF MS) analysis. The obtained data were analyzed using Principal Component Analysis (PCA) and other statistical analysis methods, and differential metabolites were further figured out. RESULTS: Compared with FY,sucrose esters (such as sibiricoses A5 and tenuifoliside B) and oligosaccharides (such as tenuifoliose K) in JY1 and FY2 contributed more to the separation of Polygala tenuifolia varieties in the PCA score plot. Compared with JYl, The sugar esters (such as tenuifoliside B and tenuifoliside A) and oligosaccharides( such as tenuifoliose A) in the FY2 also contributed more to the separation of Polygala tenuifolia varieties in the PCA score plot. In addition, the relative contents of sibiricaxanthone A,3,6'-disinapoly sucrose and senegin III showed significant differences among FY, JY1 and FY2. CONCLUSION: As new Polygala tenuifolia varieties, JY1 and FY2 had certain differences and respective advantages on the chemical composition compared with FY,which could provide data support for the directional breeding of Polygala tenuifolia based on the contents of some active compounds.

Preparation and physicochemical characterization of T-OA PLGA microspheres.

As the carrier of water-insoluble drugs, microspheres can play a role in increasing solubility and delaying releasing essence. The objective of this study was to improve the solubility and to delay the release of a newly discovered antitumor compound 3ß-hydroxyolea-12-en-28-oic acid-3, 5, 6-trimethylpyrazin-2-methyl ester (T-OA). Early-stage preparation discovery concept (EPDC) was employed in the present study. The preparation, physicochemical characterization, and drug release properties of PLGA microspheres were evaluated. T-OA-loaded PLGA microspheres were prepared by an oil-in-water (O/W) emulsification solvent evaporation method. Characterization and release behaviors of the T-OA PLGA microspheres were evaluated by X-ray diffract (XRD), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and high performance liquid chromatography (HPLC). The results demonstrated that T-OA-loaded PLGA microspheres could be successfully obtained through solvent evaporation method with appropriate morphologic characteristics and high encapsulation efficiency. The XRD analysis showed that T-OA would be either molecularly dispersed in the polymer or distributed in an amorphous form. The DSC and FTIR analysis proved that there were interactions between T-OA and PLGA polymer. SEM observations displayed the morphology of the microspheres was homogeneous and the majority of the spheres ranged between 50 and 150 µm. The drug release behavior of the microspheres in the phosphate buffered saline medium exhibited a sustained release and the duration of the release lasted for more than 23 days, which was fit with zero-order release pattern with r2 = 0.9947. In conclusion, TOA-loaded PLGA microspheres might hold great promise for using as a drug-delivery system in biomedical applications.

Phenolic constituents from Rheum nobile and their antioxidant activity.

Although the rhizomes of Rheum nobile Hook. f. et Thoms (Polygonaceae) are widely used in Tibetan medicine, no previous investigations regarding the biological activities and rarely chemical constituents of this plant have been reported. As part of an ongoing search for novel bioactive agents, a phytochemical investigation of R. nobile led to the isolation of two new compounds Rheumone B (1) and piceatannol-4'-O-ß-D-(6″-O-acetyl)-glucoside (2), together with 15 known compounds by gel filtration over Sephadex LH-20 and preparative HPLC. Their structures were determined by combined spectroscopic methods. Compounds 1-10 were evaluated for their ability to scavenge 2,2-diphenyl-1-picrylhydzyl (DPPH) radical and compounds 7-10 showed relatively strong scavenging abilities with IC50 values from 2.76 µM to 11.80 µM. In conclusion, naphthalene glycosides, stilbene glycosides, flavanols, especially anthraquinones are main chemical constituents of this plant. The ability to scavenge DPPH radical of compound 8 was the highest among compounds 1-10.