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Aim:WSS25 is a sulfated polysaccharide extracted from the rhizome of Gastrodia elata BI, which has been found to bind to bone morphogenetic protein 2 (BMP-2) in hepatocellular cancer cells. Since BMP-2 may regulate both osteoclasts and osteoblasts, here we investigated the effects of WSS25 on osteoclastogenesis in vitro and bone loss in ovariectomized mice.Methods:RAW264.7 cells or mouse bone marrow macrophages (BMMs) were treated with RANKL to induce osteoclastogenesis, which was assessed using TRAP staining, actin ring formation and pit formation assays, as well as bone resorption assay. Cell viability was detected with MTT assay. The mRNA levels of osteoclastogenesis-related genetic markers (TRAP, NFATc1, MMP-9 and cathepsin K) were detected using RT-PCR, while the protein levels of p-Smad1/5/8 and Id1 were measure with Western blotting. WSS25 was administered to ovariectomized mice (100 mg·kg−1·d−1, po) for 3 months. After the mice were euthanized, total bone mineral density and cortical bone density were measured.Results:In RAW264.7 cells and BMMs, WSS25 (2.5, 5, 10 μg/mL) did not affect the cell viability, but dose-dependently inhibited RANKL-induced osteoclastogenesis. Furthermore, WSS25 potently suppressed RANKL-induced expression of TRAP, NFATc1, MMP-9 and cathepsin K in RAW264.7 cells. Treatment of RAW264.7 cells with RANKL increased BMP-2 expression, Smad1/5/8 phosphorylation and Id1 expression, which triggered osteoclast differentiation, whereas co-treatment with WSS25 or the endogenous BMP-2 antagonist noggin suppressed the BMP-2/Smad/Id1 signaling pathway. In RAW264.7 cells, knockdown of Id1 attenuated RANKL-induced osteoclast differentiation, which was partially rescued by Id1 overexpression. In conformity to the in vitro experiments, chronic administration of WSS25 significantly reduced the bone loss in ovariectomized mice.Conclusion:WSS25 inhibits RANKL-induced osteoclast formation in RAW264.7 cells and BMMs by blocking the BMP-2/Smad/Id1 signaling pathway. WSS25 administration reduces bone loss in ovariectomized mice, suggesting that it may be a promising therapeutic agent for osteoporosis. [ABSTRACT FROM AUTHOR]

Abstract Sulfated derivatives of the galactomannan family have multiple biological activities. In the present study, sulfated galactomannan from fenugreek gum (SFG) were prepared by chlorosulfuric acid/pyridine (CSA/Py) method. In order to acquire the derivatives with the highest degree of substitution (DS), the experiment optimum conditions of the sulfation were determined by Box-Behnken design (BBD). The quadratic regression model analysis confirmed that reaction time was the most significant parameter for the impact of DS. Under the selected conditions, maximum DS was obtained as 0.490. Results of FT-IR and X-ray photoelectron spectroscopy (XPS) indicated the present of SO 3 − group. In 13C NMR spectroscopy, the original C-6 peaks did not disappear completely and the newly peaks appeared at δ 63.2 and 64.0 illustrating the incomplete substitution were predominantly on C-6 position. After sulfation, size exclusion chromatography combined with multi-angle laser light scattering (SEC-MALLS) detected all the weight average molecular mass (M W) of sulfated derivatives decrease rapidly. The introduction of the electrostatic interaction of negatively charged SO 3 − groups and the decrease in M W could have a significant effect on its biological activity. Highlights • Sulfated galactomannan of fenugreek gum is synthesized and characterized in the present work. • Sulfation conditions are optimized by response surface methodology with a maximum degree of substitution 0.490. • Structure characterization illustrated the introduction of SO 3 − and the substitution at C-6. • All sulfated fenugreek gum derivatives showed a rapidly decrease in weight average molar mass. [ABSTRACT FROM AUTHOR]

Use of microwave radiation is one of the most potential techniques in polysaccharide derivatives synthesis due to its advantages such as higher yields, milder reaction conditions and shorter reaction times. This study was aimed at producing sulfated polysaccharides by microwave irradiation using polar reagents and solvent. Six sulfated Artemisia sphaerocephala polysaccharide derivatives (SPAS) were obtained by using chlorosulfuric acid/pyridine method in the reaction duration range of 15–300 min at a fixed microwave power of 100W. Synthesis of SPAS under microwave filed was highly effective and characterized by time saving and high degrees of substitution (DS). The chemical structure of SPAS was confirmed by FT-IR and 13 C NMR that sulfation had occurred and the substitution position was mainly at C-6. Size-exclusion chromatograph combined with multi-angle laser photometer (SEC-MALLS) data indicated that short reaction time minimized acid-catalyzed degradation of polysaccharide chains. AFM observation demonstrated that low- M W derivative exhibited aggregation of polysaccharide chain as irregular spherical lumps. In anti-tumor activity assays, sulfation of PAS significantly improved the capacity to inhibit tumor cells growth by arresting the cell cycle progress in specific phases. It could be concluded that M W and molecular mass distribution had much greater influence on anti-tumor activities of SPAS against human non-small cell lung cancer A549 cells, human hepatocellular carcinoma HepG2 cells and human cervical cancer Hela cells. Microwave-assisted synthesis might possess widespread application in preparation of new polysaccharide derivatives with rich variety of structural features and bioactivities. [ABSTRACT FROM AUTHOR]

In this study, a sulfated Artemisia sphaerocephala polysaccharide (ASPs) was prepared and its antitumor activity was evaluated in tumor cells and Hepatoma 22 (H22) tumor-bearing mice. In vitro experiments, ASPs significantly inhibited the growth of HepG2 and Hela cells with the IC 50 values of 172.03 and 161.42 μg/mL, respectively. Moreover, no direct cytotoxicity against mouse fibroblast L929 normal cells was observed in vitro. After oral administration for 12 days, the tumor growth was significantly suppressed by ASPs at the doses of 200 mg/kg (inhibition rate of 60.85%). Results of tumor histological morphology and cell cycle analysis showed that ASPs could arrest H22 cells at S phase and promote cell apoptosis. Additionally, immunohistochemical analysis demonstrated that ASPs caused the down-regulation of mutant p53 protein expression in a dose-dependent manner. Therefore, these findings proposed new insight into antitumor properties of sulfated polysaccharide as a promising agent in cancer treatment. [ABSTRACT FROM AUTHOR]

In the present study, sulfated derivatives of Artemisia sphaerocephala polysaccharide (SASP) with high degree of substitution (DS) were synthesized by using 4-dimethylaminopyridine (DMAP)/dimethylcyclohexylcarbodiimide (DCC) as catalyst in homogeneous conditions. It was found that DMAP/DCC showed marked improvement in DS of sulfated samples. Compared to sulfated derivatives without catalyst, the DS of SASP increased from 0.91 to 1.28 with an increment in dosage of DMAP from 0 to 10 mg. The influence of DMAP/DCC on the DS of sulfated derivatives was depended on the content of DMAP. The effect of DMAP might be due to its strong coordination to the hydroxy group. The results of FT-IR and X-ray photoelectron spectroscopy (XPS) indicated that SO 3 − group (S 6+ , binding energy of 172.3 eV) was widely present in sulfated polysaccharide molecules. 13 C NMR results indicated that C-6 substitution was predominant for sulfated polysaccharide when compared with other positions. In the sulfation reaction, a sharp decrease in M W was observed. DMAP/DCC was an effective catalyst system in sulfated modification of polysaccharide. [ABSTRACT FROM AUTHOR]

Sulfated derivatives of galactomannan from locust bean gum (LBG) with the degree of substitution (DS) of 0.34–1.07 were synthesized using chlorosulfonic acid/pyridine (CSA/Py) method. Box–Behnken design (BBD) of response surface methodology (RSM) was employed to optimize the reaction conditions. Results of FT-IR and X-ray photoelectron spectroscopy (XPS) indicated that SO 3 H groups were widely present in sulfated LBG (SLBG). 13 C NMR result revealed that sulfation had occurred and C-6 substitution was predominant in SLBG. All sulfated samples showed a decrease in M w and more broad molar mass distribution in size exclusion chromatography combined with laser light scattering (SEC-LLS) analysis. Results of M W – 〈 S 2 〉 z 1 / 2 showed a decrease in fractal dimension ( d f ) value. Laser light scattering results also showed a conformation transition from a compact chain conformation of branched clusters to a random coil conformation of SLBG. Compared to LBG and SLBG with low DS and molecular weight, SLBG 2 exhibited an internal structure of random coil with a DS of 1.07. DS and molecular weight had great influence on its conformation in aqueous solution. Our results confirmed that the degradation of polysaccharide and SO 3 H groups improved significantly the stiffness of the chains due to the electrostatic effect. [ABSTRACT FROM AUTHOR]

Abstract: We reported here a new synthesis of C-2 and C-3 sulfated guar gum (SRSGG) with low degree of substitution (DSS) of 0.58, employing triphenylchloromethane (TrCl) as a protected precursor. The yield and DSTr (calculated from the weight of triphenylmethanol) of triphenylmethylated GG (GGTr) was 165.6% and 0.71, respectively. In addition, low ratio (1:4) of chlorosulfuric acid to pyridine (1:4) was chosen in sulfation reaction since the protecting group was slightly sensitive to acid. Results of FT-IR and 13C NMR spectroscopy indicated that C-2 and C-3 substitution was predominant but not fully sulfated in SRSGG. Size-exclusion chromatograph combined with multi-angle laser photometer (SEC-LLS) showed a decrease in molecular weight in the reaction. This might be due to the degradation in sulfation and deprotection process. Finally, we investigated the effect of structure features on the antioxidant activities in vitro. Vitro antioxidant experiments revealed that the regioselective sulfation at C-2 and C-3 and low molecular weight afforded strong antioxidant activities showing a much higher scavenging abilities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical than that by the known C-6-sulfated derivative. The antioxidant activities of sulfated polysaccharides were not a function of a single factor but a combination of molecular weight, DSS and substitution positions. [Copyright &y& Elsevier]

Abstract: It was essential to understand the chemical structure of polysaccharides for further research and biochemical or medical application of this natural biopolymer. In the present study, sulfated derivatives of guar gum with high degree of sulfation (DS) were synthesized using 4-dimethylaminopyridine (DMAP)/dimethylcyclohexylcarbodiimide (DCC) as catalyst in homogeneous conditions. The effects of the ratio of chlorosulfuric acid to pyridine, the content of catalyst and reaction temperature were investigated. Results of FT-IR, 1H and 13C NMR indicated that C-6 substitution was predominant in sulfated polysaccharide. In the sulfation reaction, a sharp decrease in M W was observed. The enhanced antioxidant activities of sulfated polysaccharides were not a function of a single factor but a combination of high DS and low molecule weight. [Copyright &y& Elsevier]

Abstract: Sulfated polysaccharides exerted potent biological property which was relative to degree of sulfation (DS), M w, substitution position and chain conformation. In the present study, sulfated derivatives of Artemisia sphaerocephala polysaccharide (ASP) with different DS were synthesized by chlorosulfuric acid/pyridine method. FT-IR and 13C NMR analysis indicated that C-6 substitution was predominant in sulfated ASP (SASP) compared with other positions at C-2. In the sulfation reaction, a sharp decrease in M w was observed in SASP. The d f values from 1.96 to 2.77 indicated that the –SO3H groups leading to the relatively expanded conformation of SASP. Antioxidant assays showed that SASP had better antioxidant activities. The data obtained in vitro models indicated that high DS and moderate M w showed the best antioxidant capacities. [Copyright &y& Elsevier]

Abstract: Sulfated polysaccharides exerted potential biological property which was relative to degree of sulfation (DS), M w, substitution position and chain conformation. In the present study, commercial guar gum was purified and its sulfated derivates with different DS and M w were synthesized. FT-IR and 13C NMR analysis indicated that C-6 substitution was predominant in sulfated samples compared with other positions. In the sulfation reaction, a sharp decrease in M w was observed. The d f values from 1.92 to 2.85 indicated that the –SO3H groups led to the relatively expanded conformation of sulfated polysaccharides. Antioxidant assays showed that sulfated polysaccharides had better antioxidant activities. The data obtained in in vitro models indicated that high DS and low M w showed the best antioxidant capacities. [Copyright &y& Elsevier]