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Screening of bioactive components from traditional Chinese medicine by immobilized beta2 adrenergic receptor coupled with high performance liquid chromatography/mass spectrometry

Traditional Chinese medicine (TCM) represents a valuable resource for lead compounds discovery. Given the complexity of TCM components, analytical methods play a key role in novel drug development. In our study, we established a high specific and reliable bio-active components screen system, where beta2 adrenergic receptor (beta2-AR) was immobilized on silica by non-covalent bonds and packed into a stainless steel column (4.6 ¡Á 50 mm, 7 mum) to form beta2-AR chromatography column. The column was further coupled with high performance liquid chromatography-time of flight tandem mass spectrometry (TOF-MS/MS). By utilizing this strategy, we successfully identified four beta2-AR-targeting compounds: tetrahydroberberine, tetrahydrocolumbamine, fumarine and corydaline from Corydalis Rhizome. The association constants between beta2-AR and tetrahydroberberine (9.04 ¡Á 104/M) as well as fumarine (4.30 ¡Á 104/M) were determined by frontal chromatography. We also found that these two compounds shared the identical binding site on immobilized beta2-AR with corresponding concentrations of 6.67 ¡Á 10?4 M and 5.88 ¡Á 10?4 M, respectively. The newly established method represents an efficient tool to identify the target specific natural compounds.

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Two new protopines argemexicaines A and B and the anti-HIV alkaloid 6-acetonyldihydrochelerythrine from Formosan Argemone mexicana

Two new protopine-type alkaloids, argemexicaine A (1) and argemexicaine B (2), along with thirteen known alkaloids, were isolated from MeOH extracts of Formosan Argernone mexicana L. (Papaveraceae). Physical and spectral analyses, particularly IR and thermo-modulated 1D and 2D NMR, were used to determine the transannular conformations of the isolated protopine-type alkaloids. The known benzo[c]phenanthridine (¡À)-6-acetonyldihydrochelerythrine (5) exhibited significant anti-HIV activity in H9 lymphocytes with EC50 and TI (Therapeutic Index) values of 1.77 mug/mL and 14.6, respectively.

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Berberis lycium Royle fruit extract mitigates oxi-inflammatory stress by suppressing NF-kappaB/MAPK signalling cascade in activated macrophages and Treg proliferation in splenic lymphocytes

Abstract: Although Berberis plant species have been advocated as immune modulators, information regarding their mechanism(s) of action is limited. Therefore, in the present study we assessed the efficacy of Berberis lycium Royle fruit extract (BLFE) in the attenuation of lipopolysaccharide (LPS)-induced oxi-inflammatory aggravation and concanavalin A (Con-A)-induced proliferation in murine peritoneal macrophages and lymphocytes, respectively. BLFE strongly suppressed production of the oxidative and inflammatory effector molecules nitric oxide (NO), reactive oxygen species (ROS), inducible nitric oxide synthase (iNOS), inflammatory cytokines (TNF-alpha/IL-6/IL-1beta/IFN-gamma) as well as chemokines (MCP-1 and RANTES), with a concomitant enhancement in heme oxygenase-1 (HO-1) and IL-10 levels. Subsequent mechanistic analysis revealed that BLFE strongly inhibited the phosphorylation of IkappaBalpha as well as MAPKs such as extracellular signal-regulated kinase (ERK), p38 MAPK, and c-Jun NH2-terminal kinase (JNK), thereby directly resulting in the suppression of nuclear factor-kappaB (NF-?B) and c-Jun activation, ultimately culminating in the observed attenuation of inflammatory molecules. Additionally, BLFE appeared to mitigate Con-A-induced proliferation of Tregs (CD3+ CD4+ CD25+) thereby suggesting its modulatory effects on adaptive immune cells. UPLC?DAD?ESI-QTOF-MS/MS of BLFE revealed the presence of major bioactive phenolics and alkaloids including chlorogenic acid, rutin, catechin and quercetin 3-d-galactoside, berberine and magnoflorine, which could have synergistically contributed to the above findings. Overall, this work provides evidence that BLFE may be effective in the mitigation of inflammatory disorders, especially those associated with NF-kappaB/MAPK activation. Graphical abstract: [Figure not available: see fulltext.].

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Efficient and general protocol for Sonogashira cross-coupling reactions of maleimides

Maleimides are involved for the first time in Sonogashira cross-coupling reactions. Sonogashira conditions have been developed to allow subsequent coupling of various terminal alkynes at room temperature in moderate to excellent yields. These results open the way to innovative synthesis strategies for various difunctionalized maleimide derivatives. The reactivity of maleimides in Sonogashira cross-coupling reactions was studied. The important role of the base is demonstrated, and optimized experimental conditions are described with a large variety of alkynes. The reactivity of the alkynes is also discussed. This reaction provides rapid access to a wide range of new maleimide derivatives in moderate to excellent yields. Copyright

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Selective inhibitor JAK3 (by machine translation)

The present invention JAK3 relates to. a compound of formula I, I/II or a pharmaceutically acceptable salt, thereof, according to the present invention. The expression of formula # I, STR3 ,Rh, Rg, Rf, m, Re, Rd, Ra, Rb # wherein Rc, as defined in the present specification, STR3 wherein, R is as defined in the present specification. The present II. invention also relates to a pharmaceutical composition comprising a I/II compound of the above formula, or a pharmaceutically acceptable salt thereof, and, as I/II described above or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for .the treatment of inflammation, such as rheumatoid arthritis. (by machine translation)

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Determination of selected isoquinoline alkaloids from mahonia aquifolia; Meconopsis cambrica; Corydalis lutea; Dicentra spectabilis; Fumaria officinalis; Macleaya cordata Extracts by HPLC-DAD and Comparison of Their Cytotoxic Activity

Alkaloids have protective functions for plants and can play an important role in living organisms. Alkaloids may have a wide range of pharmacological activities. Many of them have cytotoxic activity. Nowadays, cancer has become a serious public health problem. Searching for effective drugs with anticancer activity is one of the most significant challenges of modern scientific research. The aim of this study was the investigation of cytotoxic activity of extracts obtained from Corydalis lutea root and herb, Dicentra spectabilis root and herb, Fumaria officinalis, Macleaya cordata leaves and herb, Mahonia aquifolia leaves and cortex, Meconopsis cambrica root and herb on FaDu, SCC-25, MCF-7, and MDA-MB-231 cancer cell lines. The cytotoxic activity of these extracts has not been previously tested for these cell lines. The aim was also to quantify selected alkaloids in the investigated extracts by High Performance Liquid Chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). Cytotoxic effect of the tested plant extracts and respective alkaloid standards were examined using human pharyngeal squamous carcinoma cells (FaDu), human tongue squamous carcinoma cells (SCC-25), human breast adenocarcinoma cell line (MCF-7), human triple-negative breast adenocarcinoma cell line (MDA-MB-231). All investigated plant extracts possess cytotoxic activity against tested cancer cell lines: FaDu, SCC-25, MCF-7, and MDA-MB-231. The highest cytotoxic activity against FaDu, SCC-25, and MCF-7 cell lines was estimated for Macleaya cordata leaf extract, while the highest cytotoxic activity against MDA-MB-231 cell line was obtained for Macleaya cordata herb extract. Differences in cytotoxic activity were observed for extracts obtained from various parts of investigated plants. In almost all cases the cytotoxic activity of investigated plant extracts, especially at the highest concentration against tested cell lines was significantly higher than the activity of anticancer drug etoposide. Our investigations exhibit that these plant extracts can be recommended for further in vivo experiments to confirm their anticancer activity.

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Novel inhibitors of urease from Corydalis govaniana Wall.

A new compound govaniadine (1), along with three known tetrahydroprotoberberine-type alkaloids caseadine (2), caseamine (3), and protopine (4), were isolated from the plant Corydalis govaniana Wall. All alkaloids 1-4 exhibited a good urease enzyme inhibition with IC50 ¡À S.E.M. values of 20.2 ¡À 3.6, 38.9 ¡À 2.8, 66.7 ¡À 1.2, and 54.1 ¡À 1.2 muM, respectively, which are comparable to the standard inhibitor, acetohydroxamic acid (IC50 = 42.0 muM). None of these compounds showed inhibition against alpha-chymotrypsin. This is the first report of urease inhibiting tetrahydroprotoberberine-type alkaloids.

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ADDITIONAL ALKALOIDS FROM Glaucium squamigerum Kar. ET Kir.

A new quaternary alkaloid, (-)-beta-N-methylisocorypalminium hydroxide, was isolated from Glaucium squamigerum KAR. et KIR. in the form of iodide.In addition to the alkaloids already known to occur in this plant, i.e.allocryptopine (the main alkaloid), protopine, chelerythrine, sanguinarine, corydine, coptisine and berberine, additional alkaloids, i.e. (-)-chelidonine, (+/-)-chelidonine (diphylline) and chelirubine were also isolated from the non-quaternary fraction, while from the fraction of quaternary alkaloids (-)-beta-N-methylcanadinium iodide (the second main alkaloid), (-)-alpha-N-methylcanadinium iodide and (-)-beta-N-methylstylopinium iodide were isolated after conversion of the salts to iodides.Further the presence of small amounts of stylopine, canadine, scoulerine, corysamine, corytuberine, magnoflorine, alpha-N-methylstylopinium hydroxide and beta-N-methyltetrahydropalmatinium hydroxide were also detected.

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Aminomethylation of Michael acceptors: Complementary radical and polar approaches mediated by dialkylzincs

Phtalimidomethyl iodide and substituted maleimidomethyl iodide were used as radical precursors in dialkylzinc-mediated radical addition to diethyl fumarate. The reactions led stereoselectively to functionalized pyrrolizidines. The radical mechanism was supported by spin-trapping experiments and rationalized by theoretical calculations. Radical additions, on the one hand, and carbozincation followed by transmetalation with copper(I), on the other, were shown to be complementary methods to achieve the formal aminomethylation of activated unsaturated compounds. Copyright

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Polymeric dibromomaleimides as extremely efficient disulfide bridging bioconjugation and pegylation agents

A series of dibromomaleimides have been shown to be very efficacious at insertion into peptidic disulfide bonds. This conjugation proceeds with a stoichiometric balance of reagents in buffered solutions in less than 15 min to give discrete products while maintaining the disulfide bridge and thus peptide conformation. The insertion is initiated by disulfide reduction using a water-soluble phosphine, tris(2-carboxyethyl)phosphine (TCEP) which allows for subsequent substitution of the two maleimide bromides by the generated thiols. Reaction of salmon calcitonin (sCT) with 2,3-dibromomaleimide (1.1 excess) in the presence of TCEP (1.1 equiv) in aqueous solution at pH 6.2 gives complete production of a single conjugate which requires no workup. A linear methoxy poly(ethylene glycol) (PEG) was functionalized via a Mitsunobu reaction and used for the successful site-specific and rapid pegylation of sCT. This reaction occurs in 15 min with a small stoichiometry excess of the pegylating agent to give insertion at the disulfide with HPLC showing a single product and MALDI-ToF confirming conjugation. Attempts to use the group in a functional ATRP polymerization initiator led to polymerization inhibition. Thus, in order to prepare a range of functional polymers an indirect route was chosen via both azide and aniline functional initiators which were converted to 2,3-dibromomaleimides via appropriate reactions. For example, the azide functional polymer was reacted via a Huisgen CuAAC click reaction to an alkyne functional 2,3-dibromomaleimide. This new reagent allowed for the synthesis of conjugates of sCT with comb polymers derived from PEG methacrylic monomers which in addition gave appropriate cloud points. This reaction represents a highly efficient polymer conjugation method which circumvents problems of purification which normally arise from having to use large excesses of the conjugate. In addition, the tertiary structure of the peptide is efficiently maintained.

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