Design, synthesis and anti-tumor activity of combretastatin A-4 derivatives / 中国药科大学学报

@#Based on the structure of combretastatin A-4 (CA-4), a microtubulin inhibitor, eight novel compounds were designed and synthesized by introducing different substituents into the benzimidazole backbone which substituted B ring of CA-4, and the structures were characterized by NMR and HRMS. Proliferation inhibition of six tumor cells including A549, HepG2, HCT-116, MCF-7, PC-3 and Siha was measured by MTT method.The effect of active compound on cell migration was evaluated by scratch test.Molecular docking technique was applied to investigate the interaction between the most active compound with the tubulin and PI3K kinases respectively.Compound 4e showed prominent inhibition against six strains of tumor cells, especially with the strongest inhibitory effect on Siha cells (IC50 = 12.18 ± 1.17 μmol/L).Moreover, compound 4e could effectively inhibit cell migration, which deserves further study.Molecular docking study showed that the binding energy to the tubulin of compound 4e was stronger than that of CA-4, and the affinity with PI3Ks displayed that the PI3Kδ subtype kinase was the strongest; its binding energy was -37.2 kJ/mol.This study lays a foundation for the development of anti-tumor drug based on PI3K and microtubulin.

Study on UPLC Fingerprint of Vaccariae Semen before and after Processing and Content Determination of Erythrine and Vaccarin / 中国药房

OBJECTIVE：Compare the fingerprint difference of Vaccariae Semen before and after processed （stir-fried），and to determine the contents of erythrine and vaccarin before and after stir-fried. METHODS ：UPLC method was adopted. The determination was performed on YMC Trait C 18 column with mobile phase consisted of acetonitrile-water （gradient elution ）at the flow rate of 0.35 mL/min. The detection wavelength was set at 219 nm，and the column temperature was 35 ℃. The sample size was 1 μL. Using vaccarin as reference，the fingerprints of Vaccariae Semen crude product and its processed product （each of 17 batches，S1-S17，CS18-CS34） were drawn. The similarity evaluation and common peak identification were carried out by Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）；cluster analysis ，principle component analysis （PCA）and factor analysis were performed by using SPSS 20.0 software. The contents of erythrine and vaccarin in Vaccariae Semen crude product and its processed product were determined by UPLC. RESULTS ：There were 5 common peaks in UPLC fingerprints of 17 batches of Vaccariae Semen crude product and its processed product. The similarities were all higher than 0.99. Among them ， 2 common peaks were identified ，i.e. erythrine ，vaccarin. Results of cluster analysis showed that S 1-S17 were clustered into one category and CS 18-CS34 were clustered into one category. Results of PCA and factor analysis showed that variance contribution rate of the first principle component was 76.418％；erythrine and vaccarin had higher loading on the first principal component （eigenvalues were 0.976 and 0.966，respectively）. The linear ranges of above 2 components were 6.437-321.832 μg/mL and 7.729-386.437 μg/mL，respectively（r＞0.999）. The limits of detection and quantitation were 0.085，0.284 ng （crude product） and 0.739， 2.465 ng （processed product ）， respectively. RSDs of precision ，reproducibility，stability（12 h）and durability tests were all lower than 3％（n＝6 or n＝5）. E-mail：1083656123@qq.com Average recoveries were 96.42％（RSD＝0.85％，n＝6）and 99.13％（RSD＝1.74％，n＝6）. The contents of the two components were 0.11％-0.20％，0.42％-0.63％（crude product ）and 0.08％-0.11％，0.34％-0.50％（processed product ）. CONCLUSIONS ：UPLC fingerprint of Vaccariae Semen crude product and its processed product are established successfully. Although the chemical constituents in Vaccariae Semen are consistent before and after stir-fried ，the contents of erythrine and vaccarin are all decreased after stir-fried.

Comparison of Component Differences in Chebulae Fructus and Chebulae Fructus Immaturus Based on Chemical Pattern Recognition and Multi-index Quantitative Analysis / 中国药房

OBJECTIVE：To provide reference for the identification of Chebulae Fructus and Chebulae Fructus Immaturus . METHODS：UPLC method was adopted. The determination was performed on Waters Cortecs T 3 C18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid solution （gradient elution ）at the flow rate of 0.35 mL/min. The column temperature was 30 ℃，and the detection wavelength was set at 270 nm. The sample size was 1 μL. Using gallic acid as reference，UPLC fingerprints of 17 batches of Chebulae Fructus and 14 batches of Chebulae Fructus Immaturus were established and their similarity was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition）. By comparing substance control ， UV absorption spectrum and related literaturs ，common peaks were identified. PCA and PLS-DA were performed by using SPSS 20.0 and SIMCA 14.1 software. The contents of main difference components in Chebulae Fructus and Chebulae Fructus Immaturus were determined by above UPLC method and compared. RESULTS ：There were 8 common peaks in UPLC fingerprint of Chebulae Fructus and Chebulae Fructus Immaturus ，i.e. chebulic acid （peak 1），gallic acid （peak 2），punicalagin A （peak 3），punicalagin B （peak 4），corilagin（peak 6），chebulagic acid （peak 7）and chebulinic acid （peak 8）. The similarities of 17 batches of Chebulae Fructus were from 0.92 to 0.99，while 14 batches of Chebulae Fructus Immaturus were all above 0.99. The similarity of control fingerprint between Chebulae Fructus and Chebulae Fructus Immaturus was 0.909. PCA demonstrated the differences between Chebulae Fructus and Chebulae Fructus Immaturus . The results of PLS-DA were consistent with those of PCA ，and the variable importance in projection （VIP）values of peak 5，4，7，3 and 2 were above 1 in the PLS-DA model. In 31 batches of samples ，the contents of gallic acid （peak 2），punicalagin A（peak 3），punicalagin B （peak 4）and chebulagic acid （peak 7）were 2.63-10.31， 5.37-44.63，8.02-60.77，44.07-162.98 mg/g；RSDs were 40.14％， 47.91％ ，53.97％ ，36.22％（n＝31）. There was statistical significance in the differences of the mentioned 4 components between Chebulae Fructus and Chebulae Fructus Immaturus 719412818@qq.com （P＜0.05）. CONCLUSIONS ：There are significant differences between Chebulae Fructus and Chebulae Fructus Immaturus gallic acid ，punicalagin A ，punicalagin B and chebulagic acid are the main difference components for identification.

Effects of Shengmai injection on platelet parameters and CD4+CD25+T cells in patients with chronic lymphocytic leukemia / 中国生化药物杂志

Objective To study the effect of Shengmai injection on the platelet parameters and CD4+CD25+cells in peripheral blood of patients with chronic lymphocytic leukemia,and to explore the optimal regimen for patients with chronic lymphocytic leukemia.Methods One hundred and twenty patients with chronic lymphocytic leukemia were enrolled in this study from January 2012 to December 2014.(D1),vincristine 4mg(d1),prednisone 60 mg/d(d1-5)were given to the CHOP regimen in the control group: cyclophosphamide 600 mg/m2(d1),doxorubicin 25 mg/m2(d12)28d a course of treatment,a total of 4 courses,the observation group of patients in the control group of patients treated on the basis of Shengmai injection,compared the two groups of patients with chemotherapy,platelet parameters and serum CD4+CD25+T cells and Th17 cells.Results After treatment,the PLT,MPV and PDW of the observation group were(215.4± 31.7),(9.5±2.5)and(16.9±2.4),respectively,which were significantly higher than those of the control group.The CD4+CD25+T cells in the observation group were(1.5±0.8)The total effective rate of the control group was 35.0％,the total effective rate was 35.0％,and the total effective rate of the control group was 35.0％.There were significant difference between the control group and the control group(P<0.05),The difference was statistically significant.Conclusion Shengmai injection combined with CHOP regimen in chronic lymphocytic leukemia patients can improve immune function,promote platelet growth,improve platelet clinical parameters,and help improve the efficiency of chemotherapy,compared with CHOP alone Program treatment is better,worthy of clinical application.

Crystal structure of a secreted lipase from Gibberella zeae reveals a novel "double-lock" mechanism

Fusarium graminearum (sexual stage: Gibberella zeae) is the causative agent of Fusarium Head Blight (FHB), which is one of the most destructive plant disease of cereals, accounting for high grain yield losses, especially for wheat and maize. Like other fungal pathogens, several extracellular enzymes secreted by G. zeae are known to be involved in host infection. Among these secreted lipases, G. zeae lipase (GZEL), which is encoded by the FGL1 gene, was demonstrated to be crucial to G. zeae pathogenicity. However, the precise mechanism of GZEL remains unclear due to a lack of detailed structural information. In this study, we report the crystal structure of GZEL at the atomic level. The structure of GZEL displays distinct structural differences compared to reported homologues and indicates a unique "double lock" enzymatic mechanism. To gain insight into substrate/inhibitor recognition, we proposed a model of GZEL in complex with substrate and the lipase inhibitor ebelactone B (based on the reported structures of GZEL homologues), which defines possible substrate binding sites within the catalytic cleft and suggests an "anti sn-l" binding mode. These results pave the way to elucidating the mechanism of GZEL and thus provide clues for the design of anti-FHB inhibitors.