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How free cationic chains promote gene transfection

How free cationic chains promote gene transfection

引用

2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者：吴奇 Department of Chemistry and Department of Physics The Chinese University of Hong Kong

After unearthing that it is indeed those cationic polyethyleneimine(PEI)chains free in the mixture of polymeric non-viral vector and DNA that promote the gene transfection,we have tried in the last several years to understand the captioned question by using a combination of chemical synthesis,physical characterization and molecular *** have studied the dynamics of the complexation between different PEI chains and plasmid DNA(pDNA)and their subsequent in-vitro gene transfection under different *** the presence of free cationic chains,our recent results showed that 1)the micro-environment of the PEI/DNA polyplexes in the intracellular space nearly remains neutral(pH～7), implying that they are not delivered and developed into the later endolysosomes;and 2)blocking either the clathrin-mediated or the caveolae-medicated endocytosis has little effect,completely different from that without free *** data and our previous results also showed that 1)for longer PEI chains(～25 kg/mol)their topology has a little effect on the gene transfection,but for short chains(～2 kg/mol)linear chains are much more effective than branched ones;and 2)free cationic chains are able to interact the membranes of the cell and different organelles,including *** everything together,we come up with a hypothesis about the role of free cationic chains in the promotion of the gene ***,It is those long cationic chains(～15 nm or longer)that interact with the anionic inner membrane signal proteins and lipids to block the intervesicular fusion between ingested polyplex-containing vesicles and earlier endosomes,and at the same time,the embedded chains weaken/destabilize the vesicle membrane so that the polyplexes escape from *** hypothesis will lead to a new direction in the development of non-viral vectors for molecular medicines,including the gene transfection.

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Preparation and quality evaluation of carnosic acid self-emulsifyied drug delivery systems

Preparation and quality evaluation of carnosic acid self-emu...

引用

2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者： GAP Dong-van.YAO Ji-hong,LI Wei-ping School of Pharmacy,Dalian Medical University,Dalian 116044,China

Aim:Canosic acid is a phenolic diterpene,which is considered to be the most important antioxidant constituent of Rosemarinus officinalis L and Salvia officinalis *** camosic acid is almost insoluble in *** absorption of CA is slow after intragastric *** absolute bioavailability of camosic acid is ***,to improve the oral bioavailability of camosic acid,the camosic acid self-emulsified delivery system was prepared and evaluated. Method:The camosic acid self-emulsified formulation was optimized based on the camosic acid solubility in different oils, surfactant and *** self-microemulsified efficiency of various combinations of emulsifier and co-emulsifier was screened by the pseudo-ternary phase *** microemulsion of size and zeta potential were *** solubility, dissolution and stability of camosic acid in self-emulsified system were evaluated by HPLC assay. Results:The optimal formulation was composed of camosic acid,olive oil,Creophor EL,Labraso,Transcutol P,with the ration of 10:30:30:10:*** diluted with water,the particle size and the zeta potential were(50.9±10.2)nm and(-5.1±0.6)mv, *** acid from SMEDDS was completely and rapidly dissolved in water,0.1M HCL and pH 6.8 *** appearance and average camosic acid content of the SMEDDS were of no significant changes after 10 days of high temperature (60℃)and temperature cycling tests(40℃,4℃). Conclusion:The solubility of camosic acid is significantly increased in optimization self-emulsified system *** particles of camosic acid microemulsion were round and homogeneous and the formulation is stable and easy to prepare.

关键词： Carnosic acid self-emulsified drug delivery systems pseudo-ternary phase diagram

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稳定的PGS/NF-kB P65 siRNA基因传递系统诱导细胞凋亡增加阿霉素抗肿瘤效应

稳定的PGS/NF-kB P65 siRNA基因传递系统诱导细胞凋亡增加阿霉素...

引用

2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者：赵月芳韩丽娜冯敏中山大学药学院

目的:利用siRNA发挥RNA干扰效应治疗癌症有着巨大的应用前景。但由于缺乏稳定、有效、安全的传递载体,基于siRNA的基因治疗难以临床应用。目前,本课题组设计了一种新型可生物降解的聚谷氨酸类阳离子聚合物载体,其具有抗血清特性,可在血... 详细信息

目的:利用siRNA发挥RNA干扰效应治疗癌症有着巨大的应用前景。但由于缺乏稳定、有效、安全的传递载体,基于siRNA的基因治疗难以临床应用。目前,本课题组设计了一种新型可生物降解的聚谷氨酸类阳离子聚合物载体,其具有抗血清特性,可在血清条件下有效携带NF-κB P65 siRNA转染肿瘤细胞,抑制P65蛋白的表达,诱导肿瘤细胞凋亡:与阿霉素联用,可以协同诱导细胞凋亡。方法:利用精胺对聚谷氨酸进行氨解,制得聚谷氨酸接枝精胺的新化合物,简称PGS。以MTT法考查PGS对HeLa细胞的毒性。PGS与siRNA在生理盐水中通过静电引力作用自组装形成纳米粒,测定PGS/siRNA复合物纳米粒的粒径及表面电位。用等温量热滴定法分别考查PGS与siRNA、BSA的结合情况。用流式细胞仪分别检测在有血清条件和无血清条件下,HeLa细胞对复合物纳米粒的摄取率。HeLa细胞经复合物转染48h后,分别用蛋白质印迹法和免疫荧光法检测细胞内P65蛋白的表达水平及定位情况,再进一步用Annexin V-FITC/PI双染法检测细胞凋亡率。PGS/siRNA复合物转染HeLa细胞48h后联用阿霉素,用双染法检测凋亡细胞率,用MTT法检测细胞存活率。结果:H-NMR及C-NMR结果显示PGS制备成功。siRNA/PGS复合物纳米粒在生理盐水中粒径分布在120-160 nm。与PEI 25 kDa相比,PGS细胞毒性低。等温量热滴定结果表明,PGS与BSA结合能力弱,不易与血清中的负电蛋白相结合。当N/P大于20时,细胞摄取率高于85%。此外,血清的存在对细胞摄取不产生显著影响。PGS传递P65 siRNA可有效下调P65的表达,转染效率与经典市售转染试剂lipofectamine 2000相当。经PGS/P65 siRNA复合物纳米粒转染的HeLa细胞凋亡率高于对照lipofectamine 2000和PEI 25 kDa。PGS/siRNA复合物转染HeLa细胞48h后再使用阿霉素,可以增加阿霉素对肿瘤细胞的抗癌效应。结论:与市售lipofectamine 2000和PEI 25 kDa相比,PGS/P65 siRNA复合物纳米粒在生理盐水及有血清介质中保持着特有的稳定性及极高的转染效率。此外,PGS/siRNA复合物转染HeLa细胞可以诱导肿瘤细胞凋亡,并增加阿霉素的抗肿瘤作用。

关键词：聚谷氨酸衍生物 NF-KB p65 siRNA 凋亡阿霉素

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An novel drug delivery system based on ethosomes-templated porous silica nanoparticles

An novel drug delivery system based on ethosomes-templated p...

引用

2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者：魏玉辉兰州大学第一医院药剂科

Porous silica nanoparticles(PSN)are universally addressed as a major candidate for use as carriers in controlled drug delivery ***,most templates used to prepare PSN are poisonous *** reduce the toxicity and improve stimulatory of surfactants,the surfactant templates must be removed from the pore channel systems before drug molecules were loaded into PSN in the traditional preparing *** fact,there is no guarantee that the surfactants will be removed completely from the pore channel systems,and this loading method(drug loading after PSN formation)is far inferior to in situ loading(drug loading and PSN formation by one step)in drug loading ability. In this study,we discovered a templating route for preparing *** is not necessary to remove the templates due to harmlessness, and it could load drug in situ(shown in Fig.1).That is ethosomes,which is an interesting and innovative bilayers vesicular system embodying ethanol in relatively high concentration,and has an excellent loading *** classic ethosomes is composed of phospholipids,ethanol(20%～45%)and *** well know,phospholipids as common nonionic surfactant display good biocompatibility,and is used to prevent and treat hyperlipemia and hypertension in clinic,so ethosomes is *** addition,the mean size of ethosomes could be changed with the optimization of ethanol concentration and phosphatidylcholine in formulation. Therefore,if ethosomes was availed of template,it is not necessary to remove the ethosomes template from the pore channel systems before drug molecules were loaded into ***,if the ethosomes loading drug were availed of templates,it is not necessary to load drug by further immersing empty PSN in drug solution. Here,we approached to synthesize this novel drug delivery system of PSN based on ethosomes-templated(Ethosomes@PSN).In situ and passive loading methods were tried to prepare Ethosomes@PSN delivery systems with terbinafine hydrochloride(TH);t

关键词： Porous silica Nanoparticle Ethosomes Terbinafine Hydrochloride

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Solid lipid nanoparticles as a drug delivery system for cancer stem cell therapy

Solid lipid nanoparticles as a drug delivery system for canc...

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2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者：孙逊四川省药学会

Numerous efforts have been made to treat the various types of cancer,however the long term and spontaneous recurrence of tumors represents a major challenge in the *** reduce such patient relapse, cancer stem cell(CSC)based therapy has been introduced as a novel concept for the treatment of malignant diseases due to its ability to eliminate tumor-initiating cells,eradicating tumor forming ***, microRNA-34a was found to inhibit CSC growth,differentiation and metastasis and can therefore be developed as a therapeutic agent,raising hope for a long term lung cancer treatment. The delivery of miR-34a into the lungs for CSC-based therapy remains uncertain,with a lot of scope for *** this study,to develop a delivery system of miR-34a for CSC therapy,we used cationic solid lipid nanoparticles(SLN)to load miR-34a to treat lung metastasis of murine B16F10-CD44+(CSC-like) *** nanoparticle system demonstrated good protection for miR-34a from degradation in the serum and shared a similar size distribution at approximately 200 run by transmission electron microscopy (TEM)and photon correlation spectroscopy(PCS).The miR-34a delivered by cationic SLN could efficiently depress CD44(a CSC maker)expression followed by induction of cell *** in vivo investigation, miR-34a loaded cationic SLN showed a passive target ability to the lung *** it was also verified to be more potent in inhibition of B16F10-CD44+-bearing tumor growth and elimination of CSC-like population in the lung without *** results indicate that dSLN-miR-34a is a strong candidate for impeding CD44-positive CSC growth and migration,a key stage in the development of a CSC-based lung cancer therapy.

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Novel temperature-sensitive liposome gel based on N-Cholesteryl hemisuccinate-O-sulfate chitosan(NCHOSC) polymer for nasal delivery system

Novel temperature-sensitive liposome gel based on N-Choleste...

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2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者： Daquan Chen~(1,2*),Kaoxiang Sun~(1,2),Hongjie Mu~1,Yan Shen~3,Qineng Ping~3,Jiasheng Tu~3,Wanhui Liu~(1,2*) 1 School of Pharmacy,Yantai University,Yantai,People's Republic of China 2 State Key Laboratory ofLongacting and Targeting Drug Delivery System,Yantai,People's Republic of China 3 College of Pharmacy,China Pharmaceutical University,Nanjing,People's Republic of China

In this study,a biomimetic polymer of blood cell membrane,N-Cholesteryl hemisuccinate-O-sulfate chitosan(NCHOSC)was synthesized and characterized.A temperatur-sensitive liposome gel based on NCHOSC polymer has been applied to nasal *** curcumin-liposome was engineered to form thermogel at body temperature and degrade in an temperature *** liposome gel with a high encapsulation efficiency of Cur was formed and improved the solubility of Cur characterized using FT-IR and *** release of Cur from liposome gel was demonstrated over 24 h,which might be attributed to inter-and/or intramolecular liposome gel formation within the gel *** Cur encapsulated in liposome gels were more stable and less toxic compared to Cur loaded in *** vitro drug release results indicated that sensitive liposome gels exhibited a constant release within 3 *** research has shed some light on a temperature-sensitive liposome gel using a biomimetic polymer NCHOSC for nasal delivery system.

关键词： NCHOSC polymer thermosensitive liposome gel nasal delivery

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Preparation and physicochemical properties study on multiple agents of TCM loaded PLGA nanoparticles

Preparation and physicochemical properties study on multiple...

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2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者： Li Yonsji,Zhang Kunchi,Shaowa Lv College of Pharmacy,Heilongjiang University of Chinese Medicine,40Heping Road,Harbin 150040,P.R.China,

The final purpose of this research is finding influencing factors of multiple agents loaded nanoparticles by study of dual agents loaded *** acid(OA)and Gentiopicroside(GEN)were found to be effective in treating hepatitis from different *** the poor water-solubility of OA and short half-life period of GEN,the bioavailabilities were not *** PLGA nanoparticles simultaneously loaded with OA and GEN were *** on six independent processing parameters (included lactide-to-glycolide(L:G)ratio of PLGA,PLGA concentration,F68 concentration,acetone-to-dichloromethane(A/D) volume ratio,aqueous phase volume,initial OA and GEN content)were investigated,response surface methodology(RSM)was used to optimize preparation to enhance the incorporation of compound substances with different properties(OA and GEN,lipophilic and hydrophilic)into PLGA *** of PLGA nanoparticles were ***:PLGA nanoparticles were prepared via solvent evaporation method.A central composite design was constructed using initial OA content(A),initial GEN content(5)and PLGA concentration(C)level as independent ***:The nanoparticles produced by optimal formulation were size(186.5±2.62nm,n=3)with polydispersity index(0.295±0.02,n=3).Nanoparticles observed by transmission electron microscopy(TEM)showed extremely spherical *** entrapment efficiencies(EE)were 53.77±1.81%for OA and 26.53±2.03%for GEN(n=3).The drug loadings(DL)were 5.76±0.26 for OA and 2.84±0.19 for GEN(n=3).Conclusion:The p-value of factor A,B,AC and BC were all less than *** indicated that effects of these factors to EE of multiple agents loaded nanoparticles were significant.

关键词： Preparation and physicochemical properties study on multiple agents of TCM loaded PLGA nanoparticles TCM

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Localized and Long-Term Delivery of docetaxel liposome modified with folate-PEG-PLGA block copolymer

Localized and Long-Term Delivery of docetaxel liposome modif...

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2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者： Lei Yang,XiaoxiaoYan,Weihua Jiang,LipengQiu,HuizhuYe,Dongkai Wang~* Department of Pharmaceutics,Shenyang Pharmaceutical University,103 Wenhua Road,Shenyang 110016,PR China

Objective The aim of this study was to develop docetaxel（DTX）liposomes modified with amphiphilic block copolymer folate-poly （ethylene glycol）-poly（D,L-lactic-co-glycolic acid）(FA-PEG-PLGA,FPGA)to achieve long circulating and tumor-targeted effect. Methods FPGA copolymer was synthesized with PEG,PLGA and *** liposomes loaded DTX（FPGA-DTX-L）and DTX liposomes（DTX-L）were prepared via emulsion solvent evaporation technique and *** cytotoxicity and hemolysis test of FPGA blank liposomes,FPGA-DTX-L and DTX-L were evaluated by MTT assay using A549 cell lines and rabbit erythrocyte *** pharmacokinetic study was performed on rats to investigate long circulation feature. Results FPGA-DTX-L were found to be spherical with mean particle size（126±17）nm,zeta potential（-35.3±10.7）mV and encapsulation efficiency（95.6±4.3%）.ICof FPGA-DTX-L on A549 cell lines was approximately 3 folds lower than that of DTX-L. No red blood cells cracked or agglomerate were observed in hemolysis test,which demonstrated the liposomes were safe enough for *** in vivo study revealed that AUCof FPGA-DTX-L was 17.7 mg·h·mL,as 2.4 and 3.8 times as the values of DTX-L and DTX solution *** important,plasma clearance（CL）of FPGA-DTX-L was 45.4%and 25.6%of the values of DTX-L and DTX solution. Conclusion From the research we concluded that FA-PEG-PLGA liposomes loaded DTX could be a promising tumor-targeted and prolonged acting drug delivery system.

关键词： Docetaxel tumor-targeted liposome folate-PEG-PLGA copolymer cytotoxicity nanotechnology

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Sirolimus solid self-microemulsifying pellets:formulation development,characterization and bioavailability evaluation

Sirolimus solid self-microemulsifying pellets:formulation de...

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2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者： Xiongwei Hu~(1,2),Chen Lin~3,Dingxiong Chen~3,Jing Zhang~1,Wei Wu~2,Hongtao Song~(1,*) 1 Department of Pharmacy,Fuzhou General Hospital of Nanjing Military Region,Fuzhou 350025,China 2 School of pharmacy,Fudan University,Shanghai 201203,China 3 Department of Chemistry,Fujian Provincial Institute for Drug Control,Fuzhou 350025,China

To enhance the dissolution and oral absorption of water insoluble drug sirolimus(SRL),self-microemulsifying pellets of SRL were developed and *** test,self-emulsifying grading test,ternary phase diagrams,and central composite design were adopted to screen and optimize the composition of liquid *** selected liquid SRL-SMEDDS formulations were prepared into pellets by extrusion-spheronization method and the optimal formulation of 1mg SRL-SMEDDS pellets capsule(1.0, 22.4,38.4,19.2,121.6,30.4 and 8.0 mg of SRL,Labrafil M1944CS,Cremophor EL,Transcutol P,MCC,Lactose and CMS-Na, respectively)was finally determinated by the feasibility of the preparing process and *** optimal SRL-SMEDDS pellets showed a significant quicker redispersion rate than the dissolution rate of commercial SRL tablets Rapamune(?)in *** droplet size and polydispersity index of the reconstituted microemulsionwas almost unchanged after *** observation and scanning electron microscopic analysis confirmed good appearance of the solid ***,XRPD,and IR analysis confirmed that there was no crystalline sirolimusin the *** study in beagle dogs showed the oral relative bioavailability of SRL-SMEDDS pellets to the commercial SRL tablets Rapamune?was about 136.9%.In conclusion,the solid SMEDDS pellets might be an encouraging strategy to improve the oral absorption of SRL and the extrusion-spheronization method was a feasible technology for the solidification of liquid SMEDDS.

关键词： sirolimus self-microemulsifying drug delivery system solidification pellets redispersibility oral bioavailiability

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Characterization of an intravenous microemulsion loaded with a paclitaxel-cholesterol complex

Characterization of an intravenous microemulsion loaded with...

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2012年中国药物制剂大会——中国药学会药剂专业委员会年会、亚洲阿登制药技术研讨会暨国际控释协会中国分会年会

作者： Yanli Ma,Yu-ling Liu~* Institute of Materia Medica,Chinese Academy of Medical Science,Beijing 100050,China

Background:We have developed the paclitaxel microemulsion which can be immediately intravenous ***,the new paclitaxel microemulsion have overcome the problems caused by cremophor EL and have improved the drug *** aim of this work was to determinate the diameter,zeta potential and the extent of the large-diameter droplet tail(>5μm). Methods:the particle size and zeta potential was measured by laser light scattering using a Zeta potential/Particle sizer nicompTM 380 ZLS(PSS,U.S.A.)and the extent of the large-diameter droplet tail(>5μm)was measured by AccusizerTM 780 optical particle sizer(PSS,U.S.A.). Results:The intensity diameter of the paclitaxel microemulsion was 156.7nm(Fig.1),and it has been reported that the particles in the 60-400nm size range can slowly leak out and accumulate in tumors via a passive mechanism referred to as the 'enhanced permeability and retention '(EPR)effect,which might be conductive to a better cancer *** emulsion has a high negative zeta potential of-42.56mv(Fig2),which reflects the physical stability by preventing coalescence of droplets upon random collisions. On the other hand,The volume-weighted,large-diameter fat globule limits of the dispersed phase,expressed as the percentage of fat residing in globules larger than 5μm(PFAT5)for a given lipid injectable emulsion,must be less than 0.05%(USP 32-NF 27),the emulsion is in the specified range with 24months(Fig 3). Conclusion:with the help of cholesterol,we successfully made paclitaxel emulsion and it is stable in 24 *** diminishing the side effects,the paclitaxel microemulsion may be an advance in clinical chemotherapy applications.

关键词： paclitaxel particle size zeta potential large-diameter droplet tail

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