2013 Vol. 44 No. 5
2013, 44(5): 385-389.
DOI: 10.11665/j.issn.1000-5048.20130501
Abstract：
According to the binding mode of rivaroxaban in complex with human factor Xa and the chemical structure of rivaroxaban, we have designed and synthesized the oxazolidinone ether compounds, which have not be reported in literatures. The structures of all the rivaroxaban derivatives synthesized were identified by IR, 1H NMR and MS. The anti-factor Xa activity of the synthesized compounds was tested. The results showed that all of the tested compounds exhibited some activity, yet were less potent than rivaroxaban.
According to the binding mode of rivaroxaban in complex with human factor Xa and the chemical structure of rivaroxaban, we have designed and synthesized the oxazolidinone ether compounds, which have not be reported in literatures. The structures of all the rivaroxaban derivatives synthesized were identified by IR, 1H NMR and MS. The anti-factor Xa activity of the synthesized compounds was tested. The results showed that all of the tested compounds exhibited some activity, yet were less potent than rivaroxaban.
2013, 44(5): 390-393.
DOI: 10.11665/j.issn.1000-5048.20130502
Abstract：
A novel route to the total synthesis of lycopene was described. The synthesis was based on the following three steps: (i)the aldol condensation between 4, 8-dimethylnona-3, 7-dienal( 2 )and propionaldehyde( 3 )led to the key intermediate, 2, 6, 10-trimethylundeca-2, 5, 9-trienal( 4 ); (ii)3, 7, 11-trimethyldodeca-1, 3, 6, 10-tetraenylphosphonate( 6 )could be prepared via the modified Wittig-Horner reaction between (4) and methylenebisphosphonic acid tetraethyl ester( 5 ); (iii)another modified Wittig-Horner reaction between( 6 )and C10-triene dialdehyde( 7 )could produce all-E-lycopene( 1 ). The overall yield was 58. 7%. This approach offers many advantages including less steps, simple operations, high-yielding compared with the conventional synthetic routes, and exhibits a potential for large-scale production.
A novel route to the total synthesis of lycopene was described. The synthesis was based on the following three steps: (i)the aldol condensation between 4, 8-dimethylnona-3, 7-dienal( 2 )and propionaldehyde( 3 )led to the key intermediate, 2, 6, 10-trimethylundeca-2, 5, 9-trienal( 4 ); (ii)3, 7, 11-trimethyldodeca-1, 3, 6, 10-tetraenylphosphonate( 6 )could be prepared via the modified Wittig-Horner reaction between (4) and methylenebisphosphonic acid tetraethyl ester( 5 ); (iii)another modified Wittig-Horner reaction between( 6 )and C10-triene dialdehyde( 7 )could produce all-E-lycopene( 1 ). The overall yield was 58. 7%. This approach offers many advantages including less steps, simple operations, high-yielding compared with the conventional synthetic routes, and exhibits a potential for large-scale production.
2013, 44(5): 394-400.
DOI: 10.11665/j.issn.1000-5048.20130503
Abstract：
Gambogic acid(GA)was treated with corresponding dibromoalkanes to form brominated compounds, which were coupled with various amines to generate the target compounds 3a - 3t . Their structures were characterized by IR, MS and 1H NMR. The effects of the target compounds on the proliferation of human HCC Bel-7402, SMMC-7721, Bel-7404, QGY-7701 and HepG2 cells were evaluated in vitro by MTT assay using GA and taxol as positive controls. Most of the GA derivatives( 3c , 3g , 3h , 3p , 3t )displayed more potent activity against liver cancer cells than GA, and the activities of 3g and 3h were even more potent than that of taxol.
Gambogic acid(GA)was treated with corresponding dibromoalkanes to form brominated compounds, which were coupled with various amines to generate the target compounds 3a - 3t . Their structures were characterized by IR, MS and 1H NMR. The effects of the target compounds on the proliferation of human HCC Bel-7402, SMMC-7721, Bel-7404, QGY-7701 and HepG2 cells were evaluated in vitro by MTT assay using GA and taxol as positive controls. Most of the GA derivatives( 3c , 3g , 3h , 3p , 3t )displayed more potent activity against liver cancer cells than GA, and the activities of 3g and 3h were even more potent than that of taxol.
Abstract：
CYP1A2 enzyme plays a crucial role in drug metabolism, and its inhibition may cause low metabolic rates and increased plasma concentrations of the drugs metabolized by CYP1A2, thus leading to drug toxicity. Therefore, distinction between CYP1A2 inhibitors and non-inhibitors becomes important topic on the early selection of new drug candidates and drug safety assessment. In this study, a CYP1A2 inhibitor-ligand library was built with 674 compounds known to possess CYP1A2 inhibitory activity. From the point of receptor-based and ligand-based view, we have built an automatic screening protocol with Pipeline Pilot, using molecular docking and pharmacophore modeling methods, so as to predict inhibitors from CYP1A2 inhibitor library quickly and accurately. The final model predicted 16 target compounds from the library, and 14 of which were CYP1A2 inhibitors. At last, we used the final model to screen the American Medicine Database, and four drugs were found to possess CYP1A2 inhibitory activity. The combination of prediction models can improve the efficiency of CYP1A2 inhibitor discovery significantly.
CYP1A2 enzyme plays a crucial role in drug metabolism, and its inhibition may cause low metabolic rates and increased plasma concentrations of the drugs metabolized by CYP1A2, thus leading to drug toxicity. Therefore, distinction between CYP1A2 inhibitors and non-inhibitors becomes important topic on the early selection of new drug candidates and drug safety assessment. In this study, a CYP1A2 inhibitor-ligand library was built with 674 compounds known to possess CYP1A2 inhibitory activity. From the point of receptor-based and ligand-based view, we have built an automatic screening protocol with Pipeline Pilot, using molecular docking and pharmacophore modeling methods, so as to predict inhibitors from CYP1A2 inhibitor library quickly and accurately. The final model predicted 16 target compounds from the library, and 14 of which were CYP1A2 inhibitors. At last, we used the final model to screen the American Medicine Database, and four drugs were found to possess CYP1A2 inhibitory activity. The combination of prediction models can improve the efficiency of CYP1A2 inhibitor discovery significantly.
2013, 44(5): 410-415.
DOI: 10.11665/j.issn.1000-5048.20130505
Abstract：
Mucoinert nanoparticles(mPEG-PLGA-NPs)was prepared to overcome the adhesion of mucus in this study and the cellular uptake of nanoparticles with different physio-chemical properties by human gastric cancer HGC-27 cells was investigated. The muco-inert activity of nanoparticles was evaluated by pig gastric mucin(PM)binding experiments. Cellular uptake of nanoparticles in HGC-27 cells was analyzed by confocal laser scanning microscope and HPLC. The results indicated that mPEG-PLGA-NPs had good mucin-inert activity. The mPEG2000-PLGA-NPs could be rapidly uptaken by HGC-27 cells. The celluar uptake of 10%mPEG2000-PLGA-NPs was 1. 7-2. 0 folds higher than PLGA-NPs and 1. 8-2. 4 folds higher than CS-PLGA-NPs during the same incubation time. The intake of diameter nanoparticles with 400 nm was only 55. 9%-70. 3% of those with 120 nm. mPEG-PLGA-NPs possess hydrophilic and near neutrally-charged surfaces that minimize mucoadhesion. mPEG-PLGA-NPs can quickly penetrate the mucus surrounding HGC-27 cells and be uptaken by cells. mPEG-PLGA-NPs are expected to be used for the treatment of mucinous carcinoma.
Mucoinert nanoparticles(mPEG-PLGA-NPs)was prepared to overcome the adhesion of mucus in this study and the cellular uptake of nanoparticles with different physio-chemical properties by human gastric cancer HGC-27 cells was investigated. The muco-inert activity of nanoparticles was evaluated by pig gastric mucin(PM)binding experiments. Cellular uptake of nanoparticles in HGC-27 cells was analyzed by confocal laser scanning microscope and HPLC. The results indicated that mPEG-PLGA-NPs had good mucin-inert activity. The mPEG2000-PLGA-NPs could be rapidly uptaken by HGC-27 cells. The celluar uptake of 10%mPEG2000-PLGA-NPs was 1. 7-2. 0 folds higher than PLGA-NPs and 1. 8-2. 4 folds higher than CS-PLGA-NPs during the same incubation time. The intake of diameter nanoparticles with 400 nm was only 55. 9%-70. 3% of those with 120 nm. mPEG-PLGA-NPs possess hydrophilic and near neutrally-charged surfaces that minimize mucoadhesion. mPEG-PLGA-NPs can quickly penetrate the mucus surrounding HGC-27 cells and be uptaken by cells. mPEG-PLGA-NPs are expected to be used for the treatment of mucinous carcinoma.
2013, 44(5): 416-420.
DOI: 10.11665/j.issn.1000-5048.20130506
Abstract：
The flowability of silicified microcrystalline cellulose was compared with a mixture of standard grade of microcrystalline cellulose and silicon dioxide in order to elucidate the flowability changes of materials due to the silicification process and effects on the preparation characteristics in the direct compression process. The flowability, angle of repose, Hausner ratio and parameters in Kawakita equation were determined. Flowability differences were analyzed using scanning electron microscopy and X-ray microanalysis. Also, model drug was selected to investigate the content uniformity under desirable compressing performance. The flowability of silicified microcrystalline cellulose was better than the corresponding physical mixture, and the mechanism of flowability improvement was different. Silicified microcrystalline cellulose had better ability to improve the content uniformity of low dosage formulations. In conclusion, silicified microcrystalline cellulose is an excellent co-processed excipient, helpful to promote and improve the tablet manufacturing level.
The flowability of silicified microcrystalline cellulose was compared with a mixture of standard grade of microcrystalline cellulose and silicon dioxide in order to elucidate the flowability changes of materials due to the silicification process and effects on the preparation characteristics in the direct compression process. The flowability, angle of repose, Hausner ratio and parameters in Kawakita equation were determined. Flowability differences were analyzed using scanning electron microscopy and X-ray microanalysis. Also, model drug was selected to investigate the content uniformity under desirable compressing performance. The flowability of silicified microcrystalline cellulose was better than the corresponding physical mixture, and the mechanism of flowability improvement was different. Silicified microcrystalline cellulose had better ability to improve the content uniformity of low dosage formulations. In conclusion, silicified microcrystalline cellulose is an excellent co-processed excipient, helpful to promote and improve the tablet manufacturing level.
2013, 44(5): 421-426.
DOI: 10.11665/j.issn.1000-5048.20130507
Abstract：
The process to load doxorubicin(DOX)was optimized using all-trans-retinoid acid(ATRA)-modified low molecular weight heparin(LMWH)nanoparticles, and the in vitro characterization of DOX-loaded micelles was investigated. LMWH-ATRA conjugates(LHR)was synthesized through amide condensation reaction between LMWH and all-trans-retinoid acid(ATRA). The DOX-loaded LHR micelles were prepared by dialysis and characterized by particle size, zeta potential, drug loading content and entrapment efficiency. In vitro release of DOX from micelles was investigated in different buffer solutions(pH 4. 5, 5. 8 and 7. 4). MCF-7 cells were used to assess the cellular uptake of DOX-loaded LHR micelles. Cell apoptosis induced by DOX-loaded micelles was evaluated in MCF-7 cells by flow cytometry. LHR conjugates could encapsulate DOX up to 18. 7%, with an entrapment efficiency of 78. 8%. The particle size of negatively charged DOX-loaded micelles was in the range of 112. 1-153. 0 nm. In vitro release of DOX was faster in acidic enviroment, indicating the drug-loaded micelles were pH-sensitive. The DOX-loaded micelles were easily uptaken by MCF-7 cells compared with free DOX, and DOX/LHR micelles could induce the apoptosis of MCF-7 cells showing cytotoxicity. DOX-loaded micelles exhibited good physico-chemical properties and anti-tumor activities in vitro, indicating that LHR ploymers were proper carriers for DOX with enhanced anti-tumor activities.
The process to load doxorubicin(DOX)was optimized using all-trans-retinoid acid(ATRA)-modified low molecular weight heparin(LMWH)nanoparticles, and the in vitro characterization of DOX-loaded micelles was investigated. LMWH-ATRA conjugates(LHR)was synthesized through amide condensation reaction between LMWH and all-trans-retinoid acid(ATRA). The DOX-loaded LHR micelles were prepared by dialysis and characterized by particle size, zeta potential, drug loading content and entrapment efficiency. In vitro release of DOX from micelles was investigated in different buffer solutions(pH 4. 5, 5. 8 and 7. 4). MCF-7 cells were used to assess the cellular uptake of DOX-loaded LHR micelles. Cell apoptosis induced by DOX-loaded micelles was evaluated in MCF-7 cells by flow cytometry. LHR conjugates could encapsulate DOX up to 18. 7%, with an entrapment efficiency of 78. 8%. The particle size of negatively charged DOX-loaded micelles was in the range of 112. 1-153. 0 nm. In vitro release of DOX was faster in acidic enviroment, indicating the drug-loaded micelles were pH-sensitive. The DOX-loaded micelles were easily uptaken by MCF-7 cells compared with free DOX, and DOX/LHR micelles could induce the apoptosis of MCF-7 cells showing cytotoxicity. DOX-loaded micelles exhibited good physico-chemical properties and anti-tumor activities in vitro, indicating that LHR ploymers were proper carriers for DOX with enhanced anti-tumor activities.
2013, 44(5): 427-431.
DOI: 10.11665/j.issn.1000-5048.20130508
Abstract：
The rotary solvent-evaporation method was successfully used to prepare the co-amorphous repaglinide-saccharin(REP-SAC)with molar ratios of 1 ∶1 and 2 ∶1, respectively. The co-amorphous solids were characte-rized by differential scanning calorimetry(DSC), infrared spectroscopy(FTIR)and powder X-ray diffraction(PXRD). The solubility and dissolution of co-amorphous REP-SAC in various buffers were tested and compared with crystalline REP, physical mixtures of REP and SAC with molar ratios of 1 ∶1 and 2 ∶1, and amorphous REP. It was observed that co-amorphous REP-SAC had significantly enhanced solubility and dissolution. Also, co-amorphous REP-SAC was proved to be maintained in amorphous state with similar chemical stability to crystalline REP.
The rotary solvent-evaporation method was successfully used to prepare the co-amorphous repaglinide-saccharin(REP-SAC)with molar ratios of 1 ∶1 and 2 ∶1, respectively. The co-amorphous solids were characte-rized by differential scanning calorimetry(DSC), infrared spectroscopy(FTIR)and powder X-ray diffraction(PXRD). The solubility and dissolution of co-amorphous REP-SAC in various buffers were tested and compared with crystalline REP, physical mixtures of REP and SAC with molar ratios of 1 ∶1 and 2 ∶1, and amorphous REP. It was observed that co-amorphous REP-SAC had significantly enhanced solubility and dissolution. Also, co-amorphous REP-SAC was proved to be maintained in amorphous state with similar chemical stability to crystalline REP.
2013, 44(5): 432-436.
DOI: 10.11665/j.issn.1000-5048.20130509
Abstract：
This study was to develop 5-aminosalicylic acid pellets preparation to achieve colon adhesive. 5-amino salicylic acid loaded pellet cores using Carbomer 940 and HPC as bioadhesive agents were prepared by extrusion spheronization. The maximum drug load, different forming agents and process parameters of the pellet cores were studied with the formability as an indicator. The bioadhesion effect of different proportions of two bioadhesive agents was investigated by in vitro adhesion experiments. The pellets core was coated with Surelease® as inner layer for waterproof and Eudragit® S100 as outer layer for pH control. The in vitro release of the 5-amino salicylic acid pellets was studied in media of different pH. The maximum drug load of the pellets was 70% and PH301 was the best forming agent. The weight ratio of Carbomer 940 to hydroxypropyl cellulose should be kept 1 ∶1 to achieve high bioadhesion. The amount of Surelease® was 16% to 20% and Eudragit® S100 was 28%. Dissolution profiles of coated pellets revealed no drug release in the artificial gastric fluid within 2 h and released less than 10% in pH 6. 0 phosphate buffer within 5 h, while the coated pellets dissolved quickly in pH 7. 4 colonic fluid with a good adhesion effect. This approach was successfully utilized in the preparation of 5-amino salicylic acid pellets with the good colonic adhesion.
This study was to develop 5-aminosalicylic acid pellets preparation to achieve colon adhesive. 5-amino salicylic acid loaded pellet cores using Carbomer 940 and HPC as bioadhesive agents were prepared by extrusion spheronization. The maximum drug load, different forming agents and process parameters of the pellet cores were studied with the formability as an indicator. The bioadhesion effect of different proportions of two bioadhesive agents was investigated by in vitro adhesion experiments. The pellets core was coated with Surelease® as inner layer for waterproof and Eudragit® S100 as outer layer for pH control. The in vitro release of the 5-amino salicylic acid pellets was studied in media of different pH. The maximum drug load of the pellets was 70% and PH301 was the best forming agent. The weight ratio of Carbomer 940 to hydroxypropyl cellulose should be kept 1 ∶1 to achieve high bioadhesion. The amount of Surelease® was 16% to 20% and Eudragit® S100 was 28%. Dissolution profiles of coated pellets revealed no drug release in the artificial gastric fluid within 2 h and released less than 10% in pH 6. 0 phosphate buffer within 5 h, while the coated pellets dissolved quickly in pH 7. 4 colonic fluid with a good adhesion effect. This approach was successfully utilized in the preparation of 5-amino salicylic acid pellets with the good colonic adhesion.
Abstract：
A capillary electrophoresis(CE)method was developed for the quantification of levocetirizine dihydrochloride enantiomeric impurities. Sulfobutyl ether-β-cyclodextrin(SBE-β-CD)and hydroxypropyl-β-cyclodextrin(HP-β-CD)were selected as chiral selectors. Parameters including concentration of chiral selectors, pH of buffer solution, concentration of background electrolyte, and voltage applied were optimized by orthogonal experiment design. The optimal condition obtained was a running buffer consisting of 40 mmol/L sodium dihydrogen phosphate, 85 mg/mL chiral selectors at pH 5. 5 and voltage of 20 kV. The LOD and LOQ were 2. 0 μg/mL and 5. 0 μg/mL for isomers of levocetirizine dihydrochloride, respectively. This method could be applied to test the enantiomeric impurities of drug substance and tablets of levocetirizine dihydrochloride. A factorial experiment was exployed to investigate the complex interaction between SBE-β-CD and HP-β-CD in the separation of cetirizine enantiomers. Results indicated that synergistic effect existed between SBE-β-CD and HP-β-CD, which was only reflected in the apparent mobility rather than in the resolution.
A capillary electrophoresis(CE)method was developed for the quantification of levocetirizine dihydrochloride enantiomeric impurities. Sulfobutyl ether-β-cyclodextrin(SBE-β-CD)and hydroxypropyl-β-cyclodextrin(HP-β-CD)were selected as chiral selectors. Parameters including concentration of chiral selectors, pH of buffer solution, concentration of background electrolyte, and voltage applied were optimized by orthogonal experiment design. The optimal condition obtained was a running buffer consisting of 40 mmol/L sodium dihydrogen phosphate, 85 mg/mL chiral selectors at pH 5. 5 and voltage of 20 kV. The LOD and LOQ were 2. 0 μg/mL and 5. 0 μg/mL for isomers of levocetirizine dihydrochloride, respectively. This method could be applied to test the enantiomeric impurities of drug substance and tablets of levocetirizine dihydrochloride. A factorial experiment was exployed to investigate the complex interaction between SBE-β-CD and HP-β-CD in the separation of cetirizine enantiomers. Results indicated that synergistic effect existed between SBE-β-CD and HP-β-CD, which was only reflected in the apparent mobility rather than in the resolution.
2013, 44(5): 442-446.
DOI: 10.11665/j.issn.1000-5048.20130511
Abstract：
The objective of this study was to separate and identify the metabolites in rat plasma and urine after oral administration of salvianolic acid C, a phenolic acid in Salvia miltiorrhiza(Danshen). Solid-phase extraction method with strong anion exchange was used to separate the metabolites from the biological matrix. High-performance liquid chromatography coupled with electrospray quadrupole time-of-flight tandem mass spectrometry(HPLC-ESI/Q-TOF MS/MS)analysis was performed to identify the metabolites. Five metabolites were identified as caffeic acid(M1), monomethyl-salvianolic acid C-monoglucuronide(M2), dimethyl-salvianolic acid C-monoglucuronide(M3), dimethyl-salvianolic acid C-monoglucuronide(M4), dimethyl-salvianolic acid C(M5), as well as the prototype of salvianolic acid C detected in plasma and urine. The results indicated that methylation and glucuronidation might be the two main metabolic pathways of salvianolic acid C.
The objective of this study was to separate and identify the metabolites in rat plasma and urine after oral administration of salvianolic acid C, a phenolic acid in Salvia miltiorrhiza(Danshen). Solid-phase extraction method with strong anion exchange was used to separate the metabolites from the biological matrix. High-performance liquid chromatography coupled with electrospray quadrupole time-of-flight tandem mass spectrometry(HPLC-ESI/Q-TOF MS/MS)analysis was performed to identify the metabolites. Five metabolites were identified as caffeic acid(M1), monomethyl-salvianolic acid C-monoglucuronide(M2), dimethyl-salvianolic acid C-monoglucuronide(M3), dimethyl-salvianolic acid C-monoglucuronide(M4), dimethyl-salvianolic acid C(M5), as well as the prototype of salvianolic acid C detected in plasma and urine. The results indicated that methylation and glucuronidation might be the two main metabolic pathways of salvianolic acid C.
Abstract：
The pharmacokinetic interaction between bencycloquidium bromide nasal spray(BCQB)and budesonide nasal spray(BUD)in human was investigated. 12 male volunteers were randomly divided into two groups according to an open, randomized, two-period, two-way crossover design. In period 1, Group I took single dose BCQB 180 μg, multiple doses BCQB(180 μg, four times daily for 6 days)and combined dose(BCQB 180 μg and BUD 256 μg); Group II took single dose BUD 256 μg, multiple doses BUD(256 μg, once daily for 7 days)and combined dose(BCQB 180 μg and BUD 256 μg), successively. After a wash-out time of 7 days, the two groups exchanged their administration processes in period 2. Plasma concentration of bencycloquidium bromide and budesonide were measured by LC-MS/MS, respectively. The pharmacokinetic parameters, cmax, AUC and tmax, were calculated and statistically analyzed. Results showed that there was no obvious pharmacokinetic interaction between BUD and BCQB when they were administered jointly to healthy volunteers.
The pharmacokinetic interaction between bencycloquidium bromide nasal spray(BCQB)and budesonide nasal spray(BUD)in human was investigated. 12 male volunteers were randomly divided into two groups according to an open, randomized, two-period, two-way crossover design. In period 1, Group I took single dose BCQB 180 μg, multiple doses BCQB(180 μg, four times daily for 6 days)and combined dose(BCQB 180 μg and BUD 256 μg); Group II took single dose BUD 256 μg, multiple doses BUD(256 μg, once daily for 7 days)and combined dose(BCQB 180 μg and BUD 256 μg), successively. After a wash-out time of 7 days, the two groups exchanged their administration processes in period 2. Plasma concentration of bencycloquidium bromide and budesonide were measured by LC-MS/MS, respectively. The pharmacokinetic parameters, cmax, AUC and tmax, were calculated and statistically analyzed. Results showed that there was no obvious pharmacokinetic interaction between BUD and BCQB when they were administered jointly to healthy volunteers.
Abstract：
Compound CPU-XT-006 is a derivative of combretastatin A-4(CA-4), one kind of microtubule protein polymerization inhibitors and vascular blocking agents. MTT assay was used to detect the effect of CPU-XT-006 on the multiplication of HUVEC induced by newborn calf serum, vascular endothelial gronth factor(VEGF)or basic fibroblast growth tactor(bFGF)respectively. Flow cytometry was used to detect the HUVEC apoptosis caused by CPU-XT-006. The expression of VEGF and bFGF protein was detected by Western blot. The results showed that the compound CPU-XT-006 could inhibit HUVEC proliferation, induce HUVEC apoptosis and reduce expression of VEGF and bFGF protein.
Compound CPU-XT-006 is a derivative of combretastatin A-4(CA-4), one kind of microtubule protein polymerization inhibitors and vascular blocking agents. MTT assay was used to detect the effect of CPU-XT-006 on the multiplication of HUVEC induced by newborn calf serum, vascular endothelial gronth factor(VEGF)or basic fibroblast growth tactor(bFGF)respectively. Flow cytometry was used to detect the HUVEC apoptosis caused by CPU-XT-006. The expression of VEGF and bFGF protein was detected by Western blot. The results showed that the compound CPU-XT-006 could inhibit HUVEC proliferation, induce HUVEC apoptosis and reduce expression of VEGF and bFGF protein.
2013, 44(5): 455-459.
DOI: 10.11665/j.issn.1000-5048.20130514
Abstract：
A polysaccharide coded as ALP1 was prepared and purified from Arctium lappa L. (A. lappa L. ) roots. The hypoglycemic activity of ALP1 was determined both in vitro and in vivo. In vitro hypoglycemic assays, ALP1 significantly inhibited the activity of α-glucosidase, and the IC50 of ALP1 was determined to be 0. 518 4 mg/mL. In vivo hypoglycemic assays, ALP1 significantly reduced fasting blood glucose level, improved glucose tolerance and islet tissue lesions, reduced nitric oxide synthetase activity, decreased reactive oxygen species and malondialdehyde levels, whereas enhancing the activity of superoxide dismutase. The study indicated that ALP1 has good hypoglycemic effect on diabetic mice, and the mechanism may be related to slowing down the absorption of carbohydrates in the intestine by inhibiting the activity of α-glucosidase and reducing the oxidative damage of diabetic mice.
A polysaccharide coded as ALP1 was prepared and purified from Arctium lappa L. (A. lappa L. ) roots. The hypoglycemic activity of ALP1 was determined both in vitro and in vivo. In vitro hypoglycemic assays, ALP1 significantly inhibited the activity of α-glucosidase, and the IC50 of ALP1 was determined to be 0. 518 4 mg/mL. In vivo hypoglycemic assays, ALP1 significantly reduced fasting blood glucose level, improved glucose tolerance and islet tissue lesions, reduced nitric oxide synthetase activity, decreased reactive oxygen species and malondialdehyde levels, whereas enhancing the activity of superoxide dismutase. The study indicated that ALP1 has good hypoglycemic effect on diabetic mice, and the mechanism may be related to slowing down the absorption of carbohydrates in the intestine by inhibiting the activity of α-glucosidase and reducing the oxidative damage of diabetic mice.
Abstract：
This study investigated the influence of snogE disruption in biosynthesis pathway of nogalamycin. According to the sequence alignment of several glycosytransferases, we cloned the fragment of snogE gene sharing the highest homology with the published anthracycline antibiotics glycosyltransferase sequence. The gene disruption plasmid pSXW-2-62 was constructed and transferred into Streptomyces nogalater ATCC27451 by conjugation. Finally, we got the recombinant strain mSXW-2-71 by homologous recombination. The genotype and fermentation product of mSXW-2-71 were identified. As a result, the gene disruption plasmid was integrated into the genome and interrupted snogE gene successfully. Nogalymycin could not be detected in fermentation broth. The snogE encoding glycosyltransferase is necessary in nogalamycin biosynthesis pathway.
This study investigated the influence of snogE disruption in biosynthesis pathway of nogalamycin. According to the sequence alignment of several glycosytransferases, we cloned the fragment of snogE gene sharing the highest homology with the published anthracycline antibiotics glycosyltransferase sequence. The gene disruption plasmid pSXW-2-62 was constructed and transferred into Streptomyces nogalater ATCC27451 by conjugation. Finally, we got the recombinant strain mSXW-2-71 by homologous recombination. The genotype and fermentation product of mSXW-2-71 were identified. As a result, the gene disruption plasmid was integrated into the genome and interrupted snogE gene successfully. Nogalymycin could not be detected in fermentation broth. The snogE encoding glycosyltransferase is necessary in nogalamycin biosynthesis pathway.
Abstract：
To screen peptides with high affinity to cryptotanshinone, using cryptotanshinone as a ligand, phage clones binding to cryptotanshinone were screened with phage display 12-mer peptide library through three rounds of biopanning. After binding activity was measured by ELISA, the phage clones displaying high affinity to cryptotanshinone were selected and amplified. DNAs of phage clones were extracted and sequenced, and the coding peptide sequences were analyzed by bioinformatics. Amino acid sequences of 10 positive clones with high affinity to cryptotanshinone were identified. 614 proteins matched partially to the peptide found by homological retrieval with BLAST. The core peptide sequence VILDFGEI was deduced by this study. Peptides with high affinity to cryptotanshinone were obtained through the experiment, which provided experimental evidence and structural foundations for further study of the targets of effects and molecular mechanisms of actions of cryptotanshinone.
To screen peptides with high affinity to cryptotanshinone, using cryptotanshinone as a ligand, phage clones binding to cryptotanshinone were screened with phage display 12-mer peptide library through three rounds of biopanning. After binding activity was measured by ELISA, the phage clones displaying high affinity to cryptotanshinone were selected and amplified. DNAs of phage clones were extracted and sequenced, and the coding peptide sequences were analyzed by bioinformatics. Amino acid sequences of 10 positive clones with high affinity to cryptotanshinone were identified. 614 proteins matched partially to the peptide found by homological retrieval with BLAST. The core peptide sequence VILDFGEI was deduced by this study. Peptides with high affinity to cryptotanshinone were obtained through the experiment, which provided experimental evidence and structural foundations for further study of the targets of effects and molecular mechanisms of actions of cryptotanshinone.
2013, 44(5): 470-475.
DOI: 10.11665/j.issn.1000-5048.20130517
Abstract：
Chronic non-communicable diseases(NCDs)have become a major cause of death and disability worldwide. Therapeutic B-cell vaccines, aiming at inducing neutralizing antibodies against important mediators of such diseases, have been recognized as a new approach to treat chronic diseases. Several therapeutic B-cell vaccines have been made for the treatment of chronic diseases. This review focuses on the latest advances in the development and clinical trial of therapeutic B-cell vaccines for the treatment of hypertension, autoimmune diseases and Alzheimer disease
Chronic non-communicable diseases(NCDs)have become a major cause of death and disability worldwide. Therapeutic B-cell vaccines, aiming at inducing neutralizing antibodies against important mediators of such diseases, have been recognized as a new approach to treat chronic diseases. Several therapeutic B-cell vaccines have been made for the treatment of chronic diseases. This review focuses on the latest advances in the development and clinical trial of therapeutic B-cell vaccines for the treatment of hypertension, autoimmune diseases and Alzheimer disease
2013, 44(5): 476-481.
DOI: 10.11665/j.issn.1000-5048.20130518
Abstract：
Gene therapy holds great promise in providing new approaches for the treatment of a large number of inherited and acquired diseases, such as cancer. Yet its widespread application has been restricted by naked genes vulnerable to enzyme degradation, the rapid clearance by renal filtration, the poor cellular uptake and the inefficient escape out of endosome into cytosol. A successful gene therapy mainly relies on the development of an efficient gene delivery system. The safe and efficient delivery of therapeutic genes to host cells and subsequent release in the target compartment in a therapeutic way represents a fundamental obstacle to the clinical success of gene therapy. Here, we summarize the recent researches concerning biocompatible polymers such as chitosan, poly(amino ester)s and their derivatives as non-viral vectors for cancer gene therapy.
Gene therapy holds great promise in providing new approaches for the treatment of a large number of inherited and acquired diseases, such as cancer. Yet its widespread application has been restricted by naked genes vulnerable to enzyme degradation, the rapid clearance by renal filtration, the poor cellular uptake and the inefficient escape out of endosome into cytosol. A successful gene therapy mainly relies on the development of an efficient gene delivery system. The safe and efficient delivery of therapeutic genes to host cells and subsequent release in the target compartment in a therapeutic way represents a fundamental obstacle to the clinical success of gene therapy. Here, we summarize the recent researches concerning biocompatible polymers such as chitosan, poly(amino ester)s and their derivatives as non-viral vectors for cancer gene therapy.
2013, 44(5): 482-486.
DOI: 10.11665/j.issn.1000-5048.20130519
Abstract：
During the recent years, drug transporters have been known to play an increasingly important role in drug absorption and metabolism. Organic anion transporting polypeptides(OATPs), particularly OATP1B1, OATP1B3 and OATP2B1, are being increasingly recognized as important factors in governing the pharmacokinetics of clinical medicine because of their expression in critical tissues such as the liver, intestine and kidneys. Meanwhile, a variety of clinically used drugs have been identified as substrates of OATP transporters(e. g. many statins are substrates of OATP1B1). Some drugs may inhibit OATP transporters(e. g. cyclosporine A), causing pharmacokinetic drug-drug interactions. Moreover, genetic variability in genes encoding OATP transporters can result in marked inter-individual differences in pharmacokinetics. The aim of our work was to review the expression of OATP transporters in human and their substrates, drug-drug interactions and genetic polymorphisms, as well as their roles in drug pharmacokinetics and pharmacological efficacy.
During the recent years, drug transporters have been known to play an increasingly important role in drug absorption and metabolism. Organic anion transporting polypeptides(OATPs), particularly OATP1B1, OATP1B3 and OATP2B1, are being increasingly recognized as important factors in governing the pharmacokinetics of clinical medicine because of their expression in critical tissues such as the liver, intestine and kidneys. Meanwhile, a variety of clinically used drugs have been identified as substrates of OATP transporters(e. g. many statins are substrates of OATP1B1). Some drugs may inhibit OATP transporters(e. g. cyclosporine A), causing pharmacokinetic drug-drug interactions. Moreover, genetic variability in genes encoding OATP transporters can result in marked inter-individual differences in pharmacokinetics. The aim of our work was to review the expression of OATP transporters in human and their substrates, drug-drug interactions and genetic polymorphisms, as well as their roles in drug pharmacokinetics and pharmacological efficacy.
