Studies on flavonoids from Cirsium setosum / 中国中药杂志

To investigate the chemical constituents of ethyl acetate from Cirsium setosum, fifteen flavonoids were obtained by column chromatography on silica gel, MCI, Sephadex LH-20, and preparative HPLC. Their structures were identified as 4',5,6-trihydroxy-7-methoxyflavone(1), 4',5-dihydroxy-7,8-dimethoxyflavone(2), sorbifolin-6-O-β-glucopyranoside(3), kaempferol-7-O-α-L-rhamnoside(4), kaempferol(5), quercetin-3-O-β-D-glucosyl-7-O-α-L-rhamnoside(6), myricetin(7), myricetin-3-O-β-D-glucoside(8), 5,7- dihydroxy -3',4'- dimethoxyflavone(9), 3',4',5- trihydroxy-3,7-dimethoxyflavone(10), 3',3,4',5-tetrahydroxy-7-methoxyflavone(11), 3'-hydroxy-4',5,7-trimethoxyflavone(12), 7-hydroxy-3',4',5-trimethoxyflavone(13), 4',5-dihydroxy-2',3',7,8-tetramethoxylflavone(14), and 5-hydroxy-2',3',7,8-tetramethoxylflavone(15) by spectroscopic data analysis. All compounds were isolated from this plant for the first time.Compounds(1-15) were evaluated for their hypoglycemic activities by PTP1B enzyme model. Among them, compounds 2, 12, and 14 showed significant PTP1B inhibitory activities with IC₅₀ values of 2.54, 1.85, 2.11 μmol•L⁻¹, respectively.

In vitro anti-inflammatory and free radical scavenging activities of flavans from Ilex centrochinensis / 中国中药杂志

This study was carried out to evaluate the anti-inflammatory and free radical scavenging activities of flavans from flex centrochinensis S. Y. Hu in vitro and their structure-activity relationship. LPS-stimulated RAW 264.7 macrophage was used as inflammatory model. MTT assay for cell availability, Griess reaction for nitric oxide (NO) production, the content of TNF-alpha, IL-1beta, IL-6 and PGE, were detected with ELISA kits; DPPH, superoxide anion and hydroxyl free radicals scavenging activities were also investigated. According to the result, all flavans tested exhibited anti-inflammatory effect in different levels. Among them, compounds 1, 3, 4 and 6 showed potent anti-inflammatory effect through the inhibition of NO, TNF-alpha, IL-lp and IL-6, of which 1 was the most effective inhibitor, however, 2 and 5 were relatively weak or inactive. The order of free radical scavenging activities was similar to that of anti-inflammatory activities. Therefore, these results suggest that 3, 4 and 6, especially of 1, were,in part responsible for the anti-inflammatory and free radical scavenging activity of Ilex centrochinensis. Hydroxyl group at 4'-position of B-ring plays an important role in the anti-inflammatory and free radical scavenging capacities.

Chemical constituents in ethyl acetate extract from Rabdosia flexicaulis / 中草药

Objective: To investigate the chemical constituents from the aerial part of Rabdosia flexicaulis. Methods: Various column chromatography techniques were used to isolate and purify the compounds and their structures were identified by the spectral data. Results: Fifteen compounds were isolated and identified as rotundic acid (1), tormentic acid (2), corsolic acid (3), 23-hydroxyursolic acid (4), maslinic acid (5), teuclatriol (6), 3,6-dihydroxy-1-menthen (7), 1- hydroxypinoresinol (8), epipinoresinol (9), lariciresinol (10), caffeic acid (11), rosmarinic acid (12), 3'-O-methyl-rosmarinic acid (13), methyl 4'-O-methyl-rosmarinate (14), and methyl 3-dehydroxyl-rosmarinate (15). Conclusion: All the compounds are isolated from the species for the first time, among which compounds 1, 6, 7, 9, 13, and 14 are isolated from the plants in genus Rabdosia (Bl.) Hassk. for the first time.

Construction and functional analysis of a bispecific antibody that targets TNF-α and ED-B / 药学学报

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.

Chemical constituents in leaves of Ilex ficoidea / 中草药

Objective: To investigate the chemical constituents in leaves of Ilex ficoidea. Methods: The various chromatographic techniques were used to isolate and purify the compounds and their structures were identified by the spectral data and physicochemical properties. Results: Sixteen compounds were isolated from 95% ethanol extract and identified as friedelan-3-one (1), 4-epifriedelin (2), lupeol (3), oleanolic acid (4), 3-acetoxy-oleanoic acid (5), 3β-acetoxy-6α, 13β-dihydroxyolean-7-one (6), 2-(4-hydroxybenzyl) malic acid (7), quercetin-3-O-α-L-arabinopyranoside (8), hyperoside (9), cis-syringin (10), ethyl-O-β-L-arabinopyranoside (11), emodin (12), chrysophanol (13), physcion (14), α-spinasterol (15), and β-sitosterol (16). Conclusion: All the compounds are isolated from the species for the first time, among which compounds 1, 2, 6-8, 12, and 13 are isolated from the plants in genus Ilex L. for the first time.

Chemical constituents and their antitumor cytotoxic activity in leaves of Ilex centrochinensis / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents in leaves of Ilex centrochinensis and their antitumor bioactivity.</p><p><b>METHOD</b>Various chromatography techniques such as column chromatography on silica gel, Sephadex LH-20 and preparative HPLC were used to isolate and purify the compounds and their structures were identified by spectral data and physicochemical properties. Their antitumor effect was tested by MTT method.</p><p><b>RESULT</b>Ten compounds were isolated and identified as 1,4-benzenediol (1), (2S)-5,4'-dihydroxy-7,3'-dimethoxyflavan(2), (2S)-5,4'-dihydroxy-7-methoxyflavan (3), kaempferol (4), quercetin (5), naringenin (6), ursolic acid (7), uvaol (8), oleanolic acid (9) and beta-sitosterols (10).</p><p><b>CONCLUSION</b>Compounds 1-5, 7, 8 were isolated from the species for the first time, among which compounds 1-3 were isolated from the Ilex genus for the first time. Compounds 2 and 3 showed strong cytotoxic activity against Huh7 cell lines with IC50 values of 8.98, 13.04 mg x L(-1), respectively. Compounds 7-9 exhibited weak cytotoxic activity against Caco-2 cell lines with IC50 values of 28.52, 38.28, 33.04 mg x L(-1), respectively.</p>

Simultaneous determination of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill by RP-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>A new method for the simultaneous quantitative determination of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill using reversed phase high performance liquid chromatographic method had been developed.</p><p><b>METHOD</b>The optimum chromatographic conditions were as follows: Agilent Zorbax SB - C18 column (4.6 mm 250 mm, 5 m), acetonitrile-H2O (containing 6 mmol L(-1) KH2PO4, pH 4.6) gradient elution; as a detection wavelength of 343 nm.</p><p><b>RESULT</b>The calibration curves of geniposide, baicalin and berberine were linear at the ranges of 4.50-110.00, 5.00-153.00, 6.40-191.00 mg L(-1), respectively. The limits of detection of the method were 0.77 ng for geniposide, 1.53 ng for baicalin and 1.43 ng for berberine hydrochloride. The recoveries of the method were 104.44% (RSD 1.79% ) for geniposide, 96.98% (RSD 1.76%) for baicalin, 101.08% (RSD 3.1%) for berberine hydrochloride.</p><p><b>CONCLUSION</b>This method had been successfully applied to determine the content of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill.</p>