Norcantharidin induces apoptosis through autophagosome accumulation in breast cancer MDA-MB-231 cells / 中国药科大学学报

@#The aim of this study was to investigate the effect of norcantharidin (NCTD) on the proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231.Western blot was used to detect the effect of NCTD on the expression levels of apoptosis-related proteins Bax/Bcl-2, cleaved-PARP/PARP/PARP, cleved-caspase-9, cleaved-caspase-3 and MCL-1 in MDA-MB-231 cells.Also, the expression levels of autophagy-related proteins LC3-II/LC3-I, Parkin and PINK1 in MDA-MB-231 cells were measured by Western blot.Flow cytometry was used to measure the effect of NCTD on the changes of mitochondrial membrane potential and mitochondrial reactive oxygen species (ROS).The effect of NCTD on autophagy flow in cells expressing mCherry-EGFP-LC3 was detected by a confocal microscope.Moreover, the effects of NCTD combined with chloroquine (CQ) or 3-methyladenine (3-MA) on the apoptosis of MDA-MB-231 cells were detected by flow cytometry.The results showed that NCTD significantly increased the expression levels of Bax/Bcl-2, cleaved-PARP/PARP, cleaved-caspase-9, cleasved-caspase-3 and LC3-II/LC3-I proteins, and promoted the mitochondrial translocation of Parkin, and blocked the autophagic flow in MDA-MB-231 cells. Moreover, NCTD combined with CQ accelerated apoptosis, while NCTD combined with 3-MA decreased apoptosis.These results suggest that NCTD can induce autophagy accumulation and lead to apoptosis of MDA-MB-231 cells.

Research progress in new formulations of norcantharidin / 药学实践杂志

Objective To review the research progress in new formulations of norcantharidin. Methods The foreign and domestic literature search in the new formulations of norcantharidin was conducted. The research and development of norcantharidin formulations were summarized and commented. Results The drug delivery systems, such as microspheres, nanoparticles, liposomes, and microemulsions, have great development potential as the new formulations for norcantharidin. Conclusion Norcantharidin is an excellent anti-tumor drug. The traditional injections and tablets have serious side effects in clinical application. The new formulations reduced the renal and urinary toxicity and side effects. Those formulations provided better therapeutic effects as target medication. Therefore, the new norcantharidin formulations have great development prospects.

Mitochondrial targeting function of norcantharidin TPP-PEG-PCL nanomicelles promotes apoptosis of liver tumor cells / 中草药

Objective: To prepare norcantharidin TPP-PEG-PCL nanomicelles and study its release in vitro, intracellular transport and promoting effect on hepatoma cell apoptosis. Methods: Thin film hydration method was used to prepare norcantharidin TPP-PEG-PCL nanomicelles, and the particle size, electric potential and microscopic electron microscopy morphological analysis were measured. At the same time, the nanomicelles were evaluated for stability, in vitro release, pharmacokinetics and critical micelle concentration. Coumarin-6 was used as a fluorescent probe to evaluate the uptake of TPP-PEG-PCL nanomicelles in liver tumor cells, lysosomal escape and mitochondrial targeting function; Under the same dosage conditions, the effect of norcantharidin TPP-PEG-PCL nanomicelles on promoting apoptosis of liver tumor cells was evaluated. Results: The cantharidin TPP-PEG-PCL nanomicelles had a particle size of (16.8 ± 0.2) nm, a Zeta potential of (14.3 ± 0.2) mV, and transmission electron microscopy images showed that nanomicelles had a regular spherical shape. The fluorescence test results showed that TPP-PEG-PCL nanomicelles can promote the cellular uptake of drugs, escape lysosomal capture, and finally target aggregation at the mitochondrial site; Cell survival rate and Hoechst staining results showed that cantharidin TPP-PEG-PCL nanomicelles had a good effect on promoting apoptosis of liver tumor cells. Norcantharidin TPP-PEG-PCL nanomicelles can significantly reduce mitochondrial membrane potential, increase intracellular ROS levels, increase pro-apoptotic protein Bcl-2, and reduce resistance. The expression of apoptotic proteins Bax and these pro-apoptotic related experimental results are significantly better than those of norcantharidin PEG-PCL nanomicelles and norcantharidin, which have statistical significance. Conclusion: Norcantharidin TPP-PEG-PCL nanomicelles have good liver tumor cell mitochondrial targeting and promote tumor cell apoptosis, and it is a potentially effective drug delivery system for targeting tumor cell mitochondria.

Preparation process of norcantharidin/tetrandrine dual loaded liposomes and their release characteristics / 中国中药杂志

In order to optimize the prescription and preparation process of norcantharidin/tetrandrine dual loaded liposomes, the dual drug loaded liposomes were prepared by film dispersion-ultrasonic method using norcantharidin-mesoporous silica nanoparticles（MSN-NCTD）and tetrandrine（Tet）. With particle size and encapsulation efficiency as comprehensive indexes, the influences of phospholipid cholesterol amount, ultrasonic time and ultrasonic power on prescription process were investigated by orthogonal test; the release characteristics of liposomes were investigated by dialysis method. The results indicated that the best prescription process of prepared norcantharidin/tetrandrine dual loaded liposomes was as follows: phospholipid-cholesterol ratio 2.5:1, ultrasonic time 4 min, ultrasonic power 40%; the encapsulation efficiency was 86.62% and 79.19%respectively for NCTD and Tet;liposomes were well-shaped under the transmission microscope, with average particle size of （207.5±3.6） nm, Zeta potential of （1.345±0.173） mV; and the 48 h cumulative release rates of NCTD and Tet were 85.14% and 85.00% respectively. The experiment results proved that the dual drug loaded liposomes prepared by film dispersion-ultrasonic method had uniform particle size, high encapsulation efficiency and sustained release characteristics.

Protective Effect of Norcantharidin on Collagen-Induced Arthritis Rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of norcantharidin (NCTD) on collagen-induced arthritis (CIA) rats.</p><p><b>METHODS</b>Sixty Sprague-Dawley(SD) rats were randomly divided into 6 groups (n=10): normal group, CIA model group(model group), NCTD low-dose group [1.35 mg/(kg•d)], NCTD middle-dose group [2.7 mg/(kg•d)], NCTD high-dose group [5.4 mg/(kg•d)] and methotrexate (MTX) group [1.8 mg/(kg/w)]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin (H&E) staining. The serum levels of interleukin (IL) 1β, IL-6, tumor necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), IL-17 and transform growth factor (TGF) β were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression of retinoid-related orphan nuclear receptorγt (RORγt) and forkhead box P3 (Foxp3) in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction.</p><p><b>RESULTS</b>MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats' ankle joints compared with the model group (P<0.05 or P<0.01). All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats (P<0.05). Only middle- and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats (P<0.05). However, NCTD has no effect on vascular endothelial growth factor (VEGF) level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group (P<0.05). The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group (P<0.05).</p><p><b>CONCLUSIONS</b>NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.</p>

Preparation and Quality Evaluation of Norcantharidin Albumin Nanoparticles / 中国药师

Objective: To prepare norcantharidin albumin nanoparticles and evaluate the physical characteristics of the albumin nanoparticles. Methods:Norcantharidin albumin nanoparticles were prepared by ultra-high pressure microfluidization technology. The average particle size and the drug entrapment efficiency of albumin nanoparticles were used as the evaluation indices. Firstly,Plackett-Burman experimental design was used to screen the formula and process variables which had significant effects on the properties of albu-min nanoparticles,and then Box-Behnken experimental design was used to optimize the variables range. The morphology,particle size distribution,zeta potential and in vitro drug release behavior were investigated. Results:The average particle size of norcantharidin al-bumin nanoparticles was (105.2 ± 30.1) nm,the PdI was 0.127,and the zeta potential was( -24.7 ± 1.9) mV. In 0.5% Tween-80 phosphate buffered saline (pH 7.4),the in vitro cumulative release of norcantharidin albumin nanoparticle suspension reached up to 81.4% in 24 h. Conclusion:The preparation technology of norcantharidin albumin nanoparticles by ultra-high pressure microfluidi-zation technology is simple and feasible. The preparation technology can be used in industrial production.

Norcantharidin enhanced cisplatin sensitivity via YAP pathway in A549 cells / 中国药理学通报

Aim To investigate the effect of cotreat-ment norcantharidin (NCTD) and cisplatin (DDP) on the cisplatin sensitivity and proliferation in A549 cells, and whether YAP molecule involved in this process. Methods A549 cells were used as a model for inves-tigating the function of NCTD and DDP in this study. The expression of caspase-3,Annexin V,YAP,CTGF and Cyr61 were detected using RT-PCR and Western blot assay. The cell growth and proliferation were as-sessed by MTT and CCK-8 assay. Moreover, the tran-scriptional activity of YAP was detected by luciferase reporter gene assays. Results YAP was required for the cell growth and proliferation. NCTD,DDP and co-treatment of NCTD and DDP depressed cell viability, inhibited cell proliferation and promoted the sensitivity of cisplatin in A549 cells. Our results showed that the higher expressed oncogene YAP activated and promoted cell proliferation in lung cancer cells. Moreover, the high concentration of NCTD or DDP significantly re-duced cell proliferation, but cotreatment of NCTD and DDP in low concentration could significantly increase the cisplatin sensitivity via YAP pathway in A549 cells. Conclusions The cotreatment of NCTD and DDP in low concentration significantly reduces the transcriptional activity and protein level of YAP, then inhibits cell proliferation and thus increases the sensi-tivity of DDP in A549 cells.

Preparation of Norcantharidin Nano-micelle and Study on Its Antitumor Effect / 中国药房

OBJECTIVE:To prepare the norcantharidin (NCTD) nano-micelle and study its antitumor effect. METHODS:NCTD nano-micelle was self-formed in water using Triblock copolymers distearyl phosphatidylethanolamine-polyethylene glycol-ma-leimide;its shape was observed,the drug-loading rate,entrapment efficiency,particle size,Zeta potential were investigated. MTT was used to investigate the cell survival rate of human lung cancer A549 cells in negative control group (Phosphate buffer solu-tion),carrier group (blank nano-micelle),positive control group (NCTD APIs,5-320 μg/mL) and NCTD nano-micelle group (NCTD,5-320 μg/mL) after acting different time (24,48,72 h). Tumor nude mice were randomly divided into blank control group,NCTD injection group(1 mg/kg),NCTD low-dose,high-dose groups(0.5,1 mg/kg),6 in each group. All mice were in-travenously injected relevant medicines in tail,once a day,for 8 weeks. Tumor size was measured every week,and tumor quality was detected after the second day of finishing administration. RESULTS:NCTD nano-micelle was round,drug-loading rate was (2.82±0.05)%,entrapment efficiency was(83.67±1.78)%,particle size was(138.6±45.8)nm,Zeta potential was -(12.75± 0.34)mV(n=6). Cell survival rate of A549 cells in carrier group had no obvious changes,and was obviously decreased in posi-tive control group and NCTD nano-micelle group,which was positively correlated with concentration and time. And the decrease degree of cell survival rate in NCTD nano-micelle group was stronger than positive control group(P<0.01). Compared with blank control group,the tumor quality of mice in 3 administration groups was reduced (P<0.05),the reduction degree in NCTD na-no-micelle high-dose group was stronger than NCTD nano-micelle injection group (P<0.05). CONCLUSIONS:NCTD nano-mi-celle is successfully prepared,which has good in vitro and in vitro anti-tumor effect on A549 cells.

Early events of norcantharidin reactive oxygen species and NF-E2-related factor 2/antioxidant response element pathway of HepG2 cells / 中国药理学与毒理学杂志

OBJECTIVE To investigate the early events of norcantharidin (NCTD) induced cell apoptosis and cell cycle arrest, the variation of reactive oxygen species (ROS) and the NF-E2-relate? dactor 2/antioxidant response element(Nrf2/ARE) pathway in human HepG2 cells. METHODS The cyto?toxicity was measured by MTT assay. Apoptosis and cell cycle was analyzed by flow cytometry. The intra toxicity ROS production was evaluated by flow cytometry analysis with DCFH-DA probe and the effect of NCTD on Nrf2/ARE pathway was detected by luciferase assay in HepG2C8 cells under the same condition. The mRNA expression of heme oxygenase-1(HO-1) and NAD(P)H: quinone oxidoreductase 1(NQO1) antioxidase gene in Nrf2/ARE pathway downstream was evaluated by quantitative real-time PCR. RESULTS No significant cytotoxicity was detected after HepG2 cells were treated with NCTD 30, 60 and 120 μmol · L- 1 for 3 and 6 h, but cellular viability was inhibited significantly by NCTD 30, 60 and 120μmol·L-1 for 24, 48 and 72 h(P<0.01). Cell apoptosis and G2/M phase arrest occurred after HepG2 cells were treated with NCTD 60μmol · L-1 for 12, 24 and 48 h. The percentage of apoptosis increased from (4.00 ± 1.98)%to (12.10 ± 1.70)%for 12 h, from (4.05 ± 0.21)%to (31.8 ± 6.50)%for 24 h, and from (3.90 ± 0.85)% to (33.30 ± 1.41)% for 48 h, respectively. The percentage of G2/M phase increased from (16.51 ± 1.58)% to (40.89 ± 0.18)% for 12 h, from (16.99 ± 1.32)% to (55.29 ± 3.99)% for 24 h, and from (14.45 ± 0.59)% to (50.66 ± 5.88)% for 48 h, respectively. Compared with cell control group, the percentage of G1 phase had a significant decrease in the group with NCTD treated at different time points(P<0.01). No significant change in ROS in HepG2 cells was detected after the treatment with NCTD 30, 60 and 120μmol · L-1 for 3, 6 and 12 h. Nrf2/ARE pathway in HepG2C8 cells was activated by NCTD 30, 60 and 120μmol·L-1 for 6 and 12 h. mRNA expression of HO-1 and NQO1 had a signifi?cant activation in HepG2 cells after treatment with NCTD 30, 60 and 120 μmol · L-1 for 6 and 12 h (P<0.05). CONCLUSION NCTD can activate Nrf2/ARE pathway in the early stage in HepG2 cells, which may inhibit the intracellular ROS production in the early stage. Activation of ROS may not be the main event in NCTD induced HepG2 cell apoptosis and G2/M phase arrest.

Induction of apoptosis of cancer stem cells by norcantharidin / 中草药

Objective: To investigate the effects of norcantharidin (NCTD) on cancer stem cells (CSC). Methods: CSC were screened from UMUC3 and HepG2 cancer cell lines by limiting dilution method. The expression of stemness gene was examined by qPCR. MTS method was used to detect the inhibitory effect of NCTD and paclitaxel (PTX) on CSC. FACS was used to detect the cell apoptosis. Results: Sphere-forming cell subpopulations were obtained from two cell lines, with much higher expression of stemness genes NANOG, OCT4, ABCG2, CD133 and CD34. Serum-release led to significant down-regulation of the stemness genes. The CSC was highly resistant to PTX, but the resistance of NCTD was low. NCTD induced apoptosis of the CSC but had no significant effect on the expression of stemness genes. Conclusion: NCTD can induce apoptosis of CSC cell, without significant effect on cell differentiation.

Determination of Norcantharidin in Norcantharidin in situ Gel by HPLC / 中国药师

Objective: To establish a method for the determination of norcantharidin in norcantharidin in situ gel .Methods:An optimal HPLC method was set up and an Agilent ZORBAX SB-C18 column (150 mm ×4.6 mm, 5μm) was adopted.The mobile phase was acetonitrile-phosphate buffer solution(1∶9, adjusting pH to 3.1 with phosphorjc acid).The flow rate was 0.8 ml· min-1 and the column temperature was 25℃.The detection wavelength was set at 210 nm and the injection volume was 20μl.Results:Norcanthari-din had a good linear relationship within the range of 0.02-1.00 mg· ml-1 (r=0.999 9).The average recovery was 97.5%and RSD was 0.98%(n=9).Conclusion:The method is accurate, simple and reproducible, which can be used for the determination of nor-cantharidin in norcantharidin in situ gel .

Research of protein phosphatase 2A inhibition by norcantharidin on radiosensitivity enhancement of nasopharyngeal carcinoma CNE1 / 中国药学杂志

OBJECTIVE: To investigate the radiosensitizing effect of PP2A inhibition on nasopharyngeal carcinoma cells and its xenografts. METHODS: Co-immunoprecipitation and flow cytometry were used to examine PP2A activity, cell cycle analysis and apoptosis of CNE1 and its xenografts, in order to explore the effect of radiosensitization in CNE1 cells. RESULTS: Cell cycle distribution and apoptosis were measured by flow cytometry 24 h after treatment. The combination of 8 Gy and 40 μmol < L-1 norcantharidin produced significantly accumulation of cells in G2/M-phase( 45.71 ±2.015)% and more apoptosis(75.63 ± 6.11)% in CNE1 cell lines compared with norcantharidin alone and to radiation alone, norcantharidin in combination with radiation produced the greatest effects on tumor growth slowing and decreasing tumor weight by 87.64% relative to vehicle in CNE1 xenografts. Our data demonstrate that inhibition of PP2 A with radiation significantly sensitizes nasopharyngeal carcinoma cell lines in vitro and in vivo. CONCLUSION: The inhibition of PP2A with norcantharidin inhibited xenograft growth obviously. Our results support further exploration of PP2A inhibition as part of radiotherapy regimens for NPC and potentially other solid tumors.

Basic Knowledge and Research Progress on Cantharidin-based Molecules / 世界科学技术-中医药现代化

This study was aimed to summarize main-bone structure of cantharidin-based small molecules and facts about three typical candidates including its origin, physical-chemical properties and general synthetic approaches. Basic chemical and pharmacological information as well as development of anti-cancer activities, which were related to cantharidin, norcantharidin and their analogues were reviewed, especially the relationship between the structures and their inhibitory activity and selectivity of serine-threonine protein phosphatases PP1, PP2A and PP2B in cancer treatment. The designs and developments of new biological cantharidin-based small molecules were also reviewed.

Liver targeting anti-tumor activity of pH-sensitive liposomes loaded with lactosyl-norcantharitin phospholipids complex / 中草药

Objective: To investigate the antitumor activity of pH-sensitive liposomes (pH-LPC-lips) loaded with lactosyl-norcantharitin (Lac-NCTD) phospholipids complex in vitro and in vivo, and the liver targeting in mice. Methods: Using blank liposomes (blank-lips and blank-pH-lips) as control, the MTT assay was used to study the cytotoxic effects of Lac-NCTD and its liposomes (Lac-lips and pH-LPC-lips) on human hepatoma carcinoma cells HepG2. HPLC assay was used to evaluate the uptake of Lac-NCTD and its liposomes in HepG2. In vivo antitumor activity of Lac-NCTD and its liposomes were evaluated in mice bearing H22 liver tumors. The hepatocyte specificity of near-infrared fluorescence dye (Cy7)-labeled pH-LPC-lips in H22 tumor-bearing mice was monitored through NIR fluorescence real-time tumor imaging instrument. Results: The pH-LPC-lips demonstrated stronger cytotoxicity against tumor cells HepG2 and easily permeated the cell membrane, compared with Lac-NCTD and Lac-lips. The results of antitumor activity in vivo showed that pH-LPC-lips displayed best tumor inhibitory effect. The optical imaging results indicated that Cy7-labeled pH-LPC-lips showed excellent hepatocyte specificity in H22 tumor-bearing mice, which could reduced the side effect, and increased the antitumor activity. Conclusion: The pH-LPC-lips could take the initiative to release at the tumor site and showed the liver-targeting. As a result, the preparation could be regarded as novel liver-targeting agent which has better antitumor effect.

Experimental study of norcantharidins effects on proliferative growth and apoptosis of human cholangiocarcinoma cell line QBC939 / 肿瘤研究与临床

Objective To study the growth effect of human cholangiocarcinoma cell line QBC939 treated by norcantharidin (NCTD) and preliminary illustrate the potential mechanism.Methods The human cholangiocarcinoma cell line QBC939 was detected by MTT assay,flow cytometry,immunocytochemistry after the treatment of NCTD in vitro.Results NCTD displayed inhibitory effect on growth of QBC939 from different doses of 0.125,0.75,2.5,10,120 μg/ml after 48 h (P ＜0.05).It was in a dose and time dependent manner.Dose-effect curve was drawn and IC50 value was (3.66±1.14) μg/ml.The flow cytometric profiles showed that the rate of cell apoptosis enhanced following increasing the concentration of NCTD[(8.6±0.4) ％,(17.6±0.3) ％,(22.9±0.4) ％,(25.5±0.9) ％ and (31.1±1.5) ％,respectively]and cells blocked in the G2/M phase after treatment with 2.5 μg/ml NCTD[(14.1±1.0) ％ and (5.7±0.3) ％].The expression of the protein caspase-3 elevated after different concentrations of NCTD co-cultured with QBC939 compare with contrast group.Conclusion NCTD has an inhibitory effect on proliferation of QBC939 cell line,and the mechanism might be related to the induction of cell apoptosis and blockade of cell cycle.

Modulation of CXCR4,and MCF-7 cells invasion potential in vitro by norcantharidin / 中国生化药物杂志

Purpose To investigate the effect and mechanism of norcantharidin(NCTD)on invasion of human breast cancer cell line MCF-7 in vitro.Methods MTT assay Was used to determine MCF-7 cell proliferation,Transwell kit was used to determine cell migration,treated with NCTD on 200,400,600 μmol/L,to detect the expression of CXCR4 and CXCL12 in the control and treatment groups by real time PCR and Western blot.Results NCTD had inhibitive effects on proliferation and invasion of human breast cancer cell line MCF-7 in a dose-dependent manner,with the IC_(50) value of 582 μmol/L at 24 h,and the expression of CXCR4 Was deCreased in NCTD groups in vitro.Conclusion NCTD in vitro inhibits invasion and metastasis of human beast cancer cell line MCF-7 throush the down regulation to the expression of CXCR4.

Preparation and Evaluation of Norcantharidin-encapsulated Liposomes Modified with a Novel CD19 Monoclonal Antibody 2E8 / 华中科技大学学报（医学）（英德文版）

In this study,norcanthridin(NCTD)-encapsulated liposomes were modified with a novel murine anti-human CD19 monoclonal antibody 2E8(2E8-NCTD-liposomes)and the targeting efficiency and specific cytotoxicity of 2E8-NCTD-liposomes to CD19+leukemia cells were evaluated.BALB/c mice were injected with 2E8 hybridoma cells to obtain 2E8 monoclonal antibody(mAb).NCTD-liposomes were prepared by using film dispersion method.2E8 mAbs were linked to NCTD-liposomes using post-incorporation technology.Flow cytometry showed that the targeting efficiency of purified 2E8 mAbs on CDI9+Nalm-6 cells was 99.93%.The purified 2E8 mAbs were conjugated with NCTD-liposomes to prepare 2E8-NCTD-liposomes whose targeting efficiency on CD19+Nalm-6 was also 95.82%.The average size of 2E8-NCTD-liposomes was 118.32 nm in diameter.HPLC showed that the encapsulation efficiency of NCTD was 46.51%.When the molar ratio of 2E8/MaI-PEG2000-DSPE reached 1:50,we obtained the liposomes with 9 2E8 molecules per liposome.The targeting efficiency of 2E8-NCTD-liposomes on CD19+leukemia cells was significantly higher than that on CD19-leukemia cells.Similarly,the targeting efficiency of the immunoliposomes was also higher than that of the NCTD-liposomes on CD 19+leukemia cells.Those results were consistent with those observed by laser scanning confocal microscopy.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay demonstrated that 2E8-NCTD-liposomes specifically killed Nalm-6 cells in a dose-and time-dependent manner.The viability of Nalm-6 cells treated by 2E8-NCTD-liposomes was significantly lower than that of Molt-3cells and it was also significantly lower than that of Nalm-6 cells treated with the same concentration of NCTD-liposomes or free NCTD.We are led to concluded that 2E8 antigen can serve as a specific targeting molecule of B lineage hematopoietic malignancies for liposome targeting,and 2E8-NCTD-liposomes can be used as a new and effective means for the treatment of B lineage hematopoietic malignancies.

Investigation on anticancer activity of nanoparticles loaded lactosyl-norcantharidin / 中草药

Objective: To investigate the anticancer activity of the novel lactosyl-norcantharidin nanoparticles (Lac-NCTD-NPs) in vivo and in vitro. Methods: The MTT method was used to study the cytotoxic effects of Lac-NCTD and Lac-NCTD-NPs on HepG2, SMMC-7721, and SGC-7901 cell lines for 12 and 48 h, respectively, and the inhibitory effects of Gal-FBS; Lac-NCTD accumulated in SMMC-7721 cells was assayed by HPLC; The in vivo anticancer activity was evaluated by the tumor-growth inhibition in H22 tumor bearing mice. Results: The in vitro studies showed that the cytotoxic effects of Lac-NCTD-NPs against HepG2 and SMMC-7721 cells were the most powerful, as well as the IC 50 was the lowest, then Lac-NCTD, and they were inhibited remarkably by Gal-FBS; As for SGC-7901 cell line, the cytotoxic effects of Lac-NCTD-NPs and Lac-NCTD were not stronger than that of NCTD, and Gal-FBS had no influence on them at all; The amount of Lac-NCTD accumulated in SMMC-7721 cells was 3. 89 μg (7.02×10-3 μmol, 1×106 cell) after treatment for 12 h; The results of the antitumor activity in vivo suggested that the inhibitory rate of Lac-NCTD-NPs on tumor weight was 63.9%, which was significantly higher than that of NCTD and Lac-NCTD groups at the same molar concentration. Conclusion: The tumor-growth is inhibited effectively by Lac-NCTD-NPs which may be a kind of novel liver-targeting agents and could strongly inhibit the tumor-growth.

Preparation of Glycyrrhetic Acid Derivatives-Modified Norcantharidin Liposome and Study on Its Liver-targeting Property in Mice / 中国药房

OBJECTIVE:To prepare glycyrrhetic acid derivatives-modified norcantharidin(NC)liposome(GDNL)and study its liver-targeting property.METHODS:Gal-GAOSt targeting molecules were synthesized to modify NC and prepare GDNL,with the parameters such as the entrapment efficiency and particle diameter,etc.investigated.Mice were enrolled to be injected with GDNL and NC water solution,respectively via vena caudalis followed by determination of NC concentration in different tissues to compute the targeting-index(TI)of GDNL in liver.RESULTS:The prepared GDNL had an entrapment efficiency of 56.29%,particle diameter of(210?20)nm and TI of 5.213 in liver.CONCLUSION:The prepared GDNL has high entrapment efficiency and remarkable liver-targeting property.

Preparation and Drug Release Characteristics in Vitro of Norcantharidin Sustained-release Tablet / 中国药房

OBJECTIVE:To optimize the formulation of norcantharidin sustained-release tablet and investigate its in vitro release characteristics. METHODS: The tablet was prepared by using hydroxy-propyl methyl cellulose (HPMC) as the matrix material. The formulation of the tablet was optimized with the amount of HPMC (A),the amount of lactose (B),the concentration of ethanol (C) and the amount of ethanol (D) as factors and with the comprehensive scores of the in vitro release rate as index. Meanwhile,the in vitro release characteristics of the optimized tablets were investigated. RESULTS: The optimal formulation of the tablet was as follows: A=12 g,B=1.5 g,C=90%,D=10 mL. At 2,6 and 12 h the cumulative release rates of the tablet reached (30.63?1.03)%,(51.70?1.51)% and (89.25?1.04)%,respectively. The mechanism for the drug release was the joint action of diffusion and bulk erosion. CONCLUSION: The optimized norcantharidin sustained-release tablet is up to the expected sustained-release efficacy.