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Table of Content

    15 March 2009, Volume 18 Issue 1
    Contents

    Graphical contents list

    Journal of Chinese Pharmaceutical Sciences
    2009, 18(1):  1-4. 
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    Perspective
    Where is the biotech industry in China headed to?
    Wei-Guo Su
    2009, 18(1):  5-6. 
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    Review
    Drug delivery via the transferrin receptor-mediated endocytosis pathway
    Qing Xia, Xiu-Wei Yang, Xiao-Da Yang*, Zhong-Ming Qian, Kui Wang
    2009, 18(1):  7-13. 
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    The membrane transferrin receptor-mediated endocytosis has been exploited for developing novel targeted drug delivery systems, which could have a variety of applications in the site-specific delivery of anticancer drugs, proteins and therapeutic genes into proliferating malignant cells that overexpress the transferrin receptors. This is achieved by coupling transferrin or monoclonal antibody to transferrin receptor with therapeutic drugs or drug delivery vesicles. The transferrin conjugates can be obtained by use of either bifunctional chemical linkers or by genetic infusion of therapeutic peptides/proteins into the structure of transferrin / and monoclonal antibody to transferrin receptor. A variety of drug carriers such as liposomes, nanoparticles and DNA-polymer complexes (i.e. polyplex and lipoplex) were used to efficiently deliver the transferrin conjugates. Use of transferrin conjugates results in improvement in drug efficacy, selectivity and drug release as well as reduction in drug toxicity. This paper reviews the basic biochemistry of transferrin and the transferrin receptor as well as the strategy for developing targeted drug delivery system.

    Full Papers
    Quality evaluation of Radix Astragali from different sources in China
    Jing Liu, Hu-Biao Chen*, Xin-Gang Du, Zhong-Zhen Zhao, Bin Wang, Yu-Ying Zhao, Tao Yi
    2009, 18(1):  14-19. 
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    Radix Astragali (Huangqi) is one of the most valuable traditional Chinese medicinal herbs. It is used in the traditional Chinese medicine to reinforce ‘qi’, and it has immunostimulant, tonic, and antioxidant activities. There are many different sources of Huangqi in the market. In this study, the quality of Huangqi was evaluated by the measurement of four flavonoids, calycosin-7-O-β-D-glucopyranoside, ononin, calycosin and formononetin with HPLC-UV, as well as astragaloside IV with HPLC-ELSD. Samples included different plant species, different places of cultivation, different ages of plants, different seasons of collection and different commercial specifications. The results showed that the contents of isoflavonoids and astragaloside IV varied significantly in different sources. Our study provided useful information for the quality evaluation of Radix Astragali.
    Development of a fingerprint of Melilotus suaveolens Ledeb by high-performance liquid chromatography
    Guo-Hua Zheng#, Gui-Hong Wang#, Mu-Qin Wu, Zhi-Jun Huang, Jun-Yan Tao*, Xiao-Yu Zhang, Qiong-Guang Zhang, Fei Xiao, Chong-Ming Li
    2009, 18(1):  20-29. 
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    A simple and accurate high-performance liquid chromatography method (HPLC) was described to generate the metabolite fingerprints of Melilotus. suaveolens Ledeb of different harvesting times and different parts of the plant. A suitable chromatographic system was established using a linear gradient elution with acetonitrile and 0.1% phosphoric acid (H3PO4) as the mobile phase and a detection wavelength of 210 nm. HPLC analysis identified 12 common peaks and 8 common fingerprint peaks among the different parts of the herb. We found that the constituents varied at different harvesting times and in different parts of the plant. The HPLC fingerprint method could be used to evaluate the quality of M. suaveolens Ledeb and Melilotus genus herbs. The same strategy can also be applied for the development of fingerprints for the quality control of other herbal medicines.
    Diastereoselective synthesis of (1S)-trans-cyclopropane-1,2-dicarboxylic acid derivatives using 1,2;4,5-di-O-isopropylidene-D-fructopyranose as the chiral auxiliary
    Yun-Feng Li, Xiang-Bao Meng*, Qing Li, Zhong-Jun Li*
    2009, 18(1):  30-32. 
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    A new procedure for the highly diastereoselective synthesis of (1S)-trans-cyclopropane derivatives using a fructose derivative as the chiral auxiliary was developed. Chloroacetylated 1,2;4,5-di-O-isopropylidene-D-fructopyranose was reacted with a tertiary amine to form the ammonium salt, which was treated with cesium carbonate to give the cyclopropane derivative through ylide intermediate. The described procedure provides an efficient method to synthesize optically pure di- or poly-substituded cyclopropanes.

    Efficient synthesis of terminal α,β-unsaturated ketones as the intermediates of the proteasome epoxyketone inhibitors via Weinreb
    Yang Lü, Xiao-Min Zou, Ke Mou, Yi-Qiu Fu, Chao Ma, Bo Zhou, Ping Xu*
    2009, 18(1):  33-36. 
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    Peptidyl epoxyketones were potential antitumor agents due to their 20S proteasome inhibitory activities. Based on their structures and special inhibitory mechanism, a series of compounds were designed by linking the epoxyketone moiety (the C-terminal pharmacophore) and the peptide backbones. To make these compounds, we used a novel method to prepare the terminal α,β-unsaturated ketone, the crucial intermediate, from Weinreb amide with satisfactory yield (62%–65%).

    Application of fuzzy analytical method in the optimization of liposome preparation by orthogonal experiments
    Peng Zhen, Xiao-Jing Fan, Liang Ma, Zheng Guan, Wei Lu, Xin Hu*
    2009, 18(1):  37-42. 
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    A new statistical method, the fuzzy analytical method, was introduced in the optimization processes of liposome preparation. It took the full advantage of the information from orthogonal experiments to obtain the optimal liposome preparation conditions by considering all the evaluation indexes. Liposomes were made by the modified reverse-phase evaporation method and the properties of liposomes including size, encapsulation efficiency and physical stability were selected as the evaluation indexes to indicate the quality of liposomes. The experimental data of these properties were analyzed by three different methods including direct observation, variance analysis and fuzzy analytical method. The optimal preparation conditions obtained from these methods were validated with further experiments. The results of all possible combinations of levels for all factors in orthogonal experiments were acquired by the fuzzy analytical method. All evaluation indexes were taken into account and the optimal preparation condition was obtained. The optimal preparation conditions from direct observation and fuzzy analytical method were different and further validation studies indicated that the optimal conditions obtained from the fuzzy analytical method were in agreement with that from traditional statistical analysis. Fuzzy analytical method avoided the problem resulted from the traditional method, in which different levels of the same factor were obtained when considering different evaluation indexes. More information could be obtained from the fuzzy analytical method and the blind area within the experimental range was eliminated. As a result, fuzzy analytical method can be applied in the optimization processes of liposome preparation.

    Preparation and evaluation of sinomenine spray
    Xin-Ru Li, Xiao-Yan Li, Yan-Xia Zhou, Yan Liu*, Ming Guo, Qing-Fen Zhu, Yuan-Chao Xie, Zhi-Yun Fan
    2009, 18(1):  43-48. 
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    We prepared and characterized the spray formulation of sinomenine for topical administration. The permeation enhancer was selected based on in vitro skin permeation studies. The antioxidant was optimized through stability studies. Pharmacokinetic and pharmacological properties were determined in order to evaluate its side effect and pharmacodynamic action. It was found that azone was the most effective permeation enhancer, and sulfocarbamide was the optimal antioxidant. Alcohol was used as the additive of the sinomenine spray. Sinomenine spray decreased arthritis index of rats induced by Freund’s complete adjuvant. The pharmacokinetic parameters of Cmax and AUC0–24 for spray were 1.17 μg/mL and 3.35 μg/h/mL, respectively, which were lower than those by oral administration. The relative bioavailability was 18.0%. Sinomenine spray was a promising topical delivery system with comparable anti-inflammatory efficacy, but possibly lower side effect.

    Determination of bergenin in rat urine, feces and tissues by RP-HPLC method
    Yan-Bin Shi*, Yan-Ping Shi, Xiao-Yun Zhang, Quan-Yi Zhao, Jing-Man Ni
    2009, 18(1):  49-54. 
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    A RP-HPLC analytical method was developed for determining bergenin in rat urine, feces and tissues. The separation was performed on a C18 analytical column with methanol-water as the mobile phase at a flow rate of 0.8 mL/min. The column temperature was 40 ºC and the eluate was monitored at 220 nm. The calibration curve for urine analysis showed a good linearity (r = 0.9989) over the range of 0.5-100 µg/mL, and the absolute recoveries were in the range of 90.0%-92.6%. A good linearity (r0.9996) for all tested tissues was obtained over the range of 0.3-150 µg/g except for lung, which had a linearity range of 0.5-150 µg/g. The absolute recoveries from tissues were higher than 63.9%. The method was successfully applied to determine the bergenin levels in several biological samples after rats were given bergenin formulation through tail vein injection. The results demonstrated that the amount of bergenin recovered from urine was about 35.36% of the total dose, and that from feces was less than 10%. Bergenin can be quickly and widely distributed in the rat, accumulating high levels in the kidney in comparison to all other tissues investigated. It was not distributed into the brain at any significant level owing to its hydrophilic property.

    An efficient method for the simultaneous determination of three flavone aglycones in Flos Chrysanthemi by acid hydrolysis and HPLC
    Fu-Gao Li, Hui Zhou, Do-Lin Sun, Ru-Wei Wang, Su Zeng, Hui-Di Jiang*
    2009, 18(1):  55-60. 
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    An efficient method for the simultaneous quantification of three flavone aglycones was developed and validated for the control of flavonoids in Flos Chrysanthemi (FC). The method employed mixed solvent of hydrochloric acid (HCl) and methanol to extract and hydrolyze flavone glycosides simultaneously from powdered FC followed by HPLC to determine the contents of three flavone aglycones, luteolin, apigenin and diosmetin, which are the major bioactive flavones in FC. The extraction and hydrolysis conditions optimized by the orthogonal tests were as follows: powdered FC was refluxed in 80% methanol (v/v) containing 2.4 M HCl for 2 h in 80 ºC water bath. The HPLC separation was performed on C18 column and detected with DAD at 344 nm. The calibration curves for luteolin, apigenin and diosmetin were linear over the ranges of 2.233-71.46 μg/mL, 1.668-53.38 μg/mL and 1.410-45.11 μg/mL, their assay recoveries were 100.3%, 103.9% and 100.4%, their intra-day repeatability (R.S.D.) were 0.68%, 0.21% and 0.38% and their inter-day repeatability were 2.6%, 1.7% and 2.0%, respectively. The developed method has been successfully utilized to assay eight species of popular FC samples, especially to analyze Hangbaiju, the well-known FC in China.
    Development of a chiral HPLC method for the analysis of naftopidil enantiomers
    Yin-Xiang Sun, Bi-Yun Huang, Mu Yuan*, Jing-Shan Shi
    2009, 18(1):  61-63. 
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    We developed a chiral HPLC method for the separation and analysis of naftopidil enantiomers. The two enantiomers of naftopidil were separated using a Chiralpak AD-H (250 mm×4.6 mm, 5 μm) column and monitored at the wavelength of 283 nm. The isocratic mobile phase consisting of hexane–isopropanol–diethylamine (85:15:0.1, v/v/v) was pumped at a flow rate of 1.0 mL/min. Under these chromatographic conditions, R-naftopidil and S-naftopidil were well separated and had good linearity in the ranges of 0.78–50 μg/mL (r = 0.9999) and 0.84–54 μg/mL (r = 0.9998), respectively. The relative standard deviations (RSD) of intra- and inter-day assays were no more than 0.5% and 0.7%, respectively. This improved method for the separation and quantitative determination of naftopidil enantiomers can be used for the quality control of synthesized naftopidil product.

    Cardioprotective effects of hydrogen sulfide and nitric oxide and their interactions during the process of myocardial ischemia in rats
    Chang-Qing Chen#, Hong Xin#, Yi-Chun Zhu, Yi-Zhun Zhu*
    2009, 18(1):  64-72. 
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    The relationship between hydrogen sulfide (H2S) and nitric oxide (NO) in myocardial infarction (MI) has not been previously reported. In the current investigation, we sought to determine the roles of both H2S and NO in MI in rats. Animals were randomly divided into 5 groups and treated with L-NG-nitro arginine methyl ester (L-NAME), sildenafil, saline, propargylglycine (PAG) and L-cysteine, respectively, for 1 week prior to performing MI surgery or sham operation. The mortality rates were lower in sildenafil and L-cysteine treated rats in the MI group. The infarct area was significantly reduced in sildenafil and L-cysteine treated rats. Moreover, plasma H2S measurements revealed that the level in the sildenafil treated group was lower than in the L-NAME treated MI group, which was consistent with an observed decrease in cystathionine gamma-lyase (CSE) enzyme activity. CSE protein expression level in the L-NAME treated MI group was significantly higher than in sildenafil treated MI group. eNOS protein content in the L-cysteine treated MI group was lower than in the PAG treated MI group and eNOS gene expression is significantly decreased in the L-cysteine treated rats. We demonstrated that endogenous H2S and NO are cardioprotective in the rat model of MI. Indeed, both the H2S-CSE and NO-NOS system appear to have a mutual down-regulation effect in MI process in our experimental rat model.
    Protective effect of xylocoside G on Aβ25-35-induced neurotoxicity in PC12 cells
    Jia Ge, Yan Yu, De-Hua Cui*, Peng-Fei Tu*
    2009, 18(1):  73-78. 
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    We studied the protective effect of xylocoside G on Aβ25-35-induced apoptosis in PC12 cells. Cell viability was analyzed by MTT assay, and apoptotic neuronal death was assessed by Hoechst 33342 staining. Flow cytometry was used to determine the apoptotic neuronal cells quantitatively. The level of intracellular reactive oxygen species (ROS) was monitored with the fluorescent probe 2’,7’-dichlorofluoresce in diacetate (DCFH-DA). We found that PC12 cells treated with 25 μM Aβ25-35 for 24 h had a significant decrease in cell viability compared with control, and the percentage of apoptotic cells was increased to 34.26%. The level of intracellular oxygen species was also increased. Co-incubation with xylocoside G (10 μM) for 24 h attenuated the damaging effect of Aβ25-35 on the cell viability, and the percentage of apoptotic cells was decreased to 22.62%. Moreover, the increase of ROS induced by Aβ25-35 was inhibited by xylocoside G (10 μM). We concluded that xylocoside G had protective effect against Aβ25-35-induced apoptosis, which might be related with the decrease of the level of ROS.

    Pharmacokinetics studies of aristolochic acid-I and -II in rats after intragastrical administration of Radix Aristolochiae and Muskone
    Pema Dolma, Dong-Hui Yang*, Jie Yu, Xuan Wang, Shao-Qing Cai
    2009, 18(1):  79-83. 
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    The pharmacokinetic parameters of aristolochic acid-I (AA-I) and -II (AA-II) in rat serum after intragastrical administration of the crude drug powders of Radix Aristolochiae (RA) and Muskone containing equal amounts of RA were compared. The pharmacokinetic profiles of AA-I and AA-II could be fitted with a two-compartment model. The elimination half time (T1/2β) of AA-I in Muskone was 1573.2 min and that of AA-I in RA was 475.8 min; T1/2β of AA-II in Muskone was 2344.8 min and that of AA-II in RA was 427.8 min. The area under the concentration-time curve (AUC) of AA-I in Muskone was 13.07 μg/h/mL and that of AA-I in RA was 3.86 μg/h/mL; AUC of AA-II in Muskone was 67.67 μg/h/mL and that of AA-II in RA was 23.93 μg/h/mL. The bioavailabilities of AA-I and AA-II in Muskone were markedly increased compared with that in RA based on the elimination half-time and AUC values.
    Notes
    Preliminary analysis of the protein contents in Pinellia ternata with different types of leaves
    Hai-Xia Wen*, Jia-Li Cai, Ming Zhang, Xing-Jia Ming, Ri-Yong Liu, Si-Yu Tang, Qing-Rui Zhou
    2009, 18(1):  84-87. 
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    Total proteins extracted from Pinellia ternata with different types of leaves have been assayed by UV absorbance, blue native polyacrylamide gel electrophoresis (BN-PAGE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We found that the total protein contents of P. ternata with the herbaceous peony and claw types of leaves were much higher than that with other types of leaves. P. ternata collected from Nanchuan County, except the ones with herbaceous peony leaves, displayed a high similarity with each other. Both UV absorbance and cluster analysis indicated that the protein content in P. ternata with the herbaceous peony leaves was less affected by habitats than that with other types of leaves. The results showed that it was necessary to increase the homogeneity of seeds for improving the protein content of P. ternata.
    News
    Instruction for Authors
    Journal of Chinese Pharmaceutical Sciences
    2009, 18(1):  89-91. 
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    Introduction of cover
    Peng-Fei Tu*
    2009, 18(1):  92-92. 
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