单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,用于基因和抗癌药物控制释放高分子材料研究,卓仁禧 院士,生物医用高分子材料教育部重点实验室,武汉大学化学与分子科学学院,卓仁禧院士_用于基因和抗癌药物控制释放高分子材料研究(minimizer)课件,1,一.高分子基因治疗载体研究,病毒载体和非病毒载体的比较,效率低,安全,容易制备，储存和质量控制,传递的DNA大小从20个碱基对到几十万个碱基对,非病毒,载体,安全隐患,DNA大小受限制,免疫原性,高效,病毒,载体,缺点,优点,载体,一.高分子基因治疗载体研究效率低安全非病毒安全隐患高效病,2,非病毒载体,阳离子脂质体,高分子,:阳离子高分子,水溶性非离子高分子,高分子基质,非病毒载体 阳离子脂质体,3,常用的聚阳离子基因载体,Poly,-(4-amino-butyl)-L-glycolic acid,S W Kim,Utah Univ,Poly-L-lysine,Polyethylenimines(PEI),Behr J P,France,Dendrimer,Szoka,UCSF,&Baker J,Univ Mich,Polyphosphoester and Polyphosphoamide,Leong K W,Johns Hopking Univ,Langer R,MIT,常用的聚阳离子基因载体Poly-(4-amino-but,4,从新的起始核合成聚酰胺胺树型高分子基因载体,从新的起始核合成聚酰胺胺树型高分子基因载体,5,Dendrimer GT5,Dendrimer GT5,6,Transfection of HEK 293 cells with pRE Luc/dendrimer GT complexes,合成的树型高分子的转染效率与目前最好的阳离子载体PEI相当，但是比PEI的毒性低得多,Cytotoxicity of dendrimer GT3GT5 measured by the MTT assay.PEI and PLL were used as controls,Transfection of HEK 293 cells,7,肝细胞靶向基因载体,肝细胞靶向基因载体,8,1.0E+04,1.0E+05,1.0E+06,1.0E+07,1.0E+08,1.0E+09,0,2,4,6,8,10,N/P,RLU/mg protein,GT5,GT5-Gal 5%,Transfection of HepG2 cells with pRE Luc,1.0E+041.0E+051.0E+061.0E+071.,9,Transfection of NIH 3T3 cells with pRE Luc,Transfection of NIH 3T3 cells,10,阳离子聚磷酸酯基因载体,阳离子聚磷酸酯基因载体,11,卓仁禧院士_用于基因和抗癌药物控制释放高分子材料研究(minimizer)课件,12,PPA-DPA,Gal-PPA,PPA-DPAGal-PPA,13,Gal-PEG-PPA,Gal-PEG-PPA,14,liver,spleen,lung,kidney,heart,10,3,10,5,10,4,10,2,RLU/mg protein,PPA,Gal-PPA,Gal-PEG-PPA,Luciferase Expression in mouse after intraportal vein injection of complexes containing 50,g plasmid,liverspleenlungkidneyheart1031,15,二.药物控释生物可降解高分子材料,研究,聚酯(聚乳酸,聚乙醇酸及其共聚物;聚己内酯,聚羟基丁酸及其共聚物),聚酸酐,聚碳酸酯及其共聚物,聚磷酸酯及其共聚物,纳米药物控制释放,温度敏感快速响应PNIPA水凝胶,Poly(lactide-co-glycolide),PLGA,Polylactide,PLA,Polyglycolide,PGA,二.药物控释生物可降解高分子材料研究Poly(lacti,16,聚碳酸酯及其共聚物,BTMC,PTMC,卓仁禧,王旭立,刘立建.苄氧基三亚甲基碳酸酯及其制备方法。中国专利,ZL 01 1 14284.7 C07D319/04,卓仁禧，王旭立，刘立建。聚（取代基三亚甲基碳酸酯）及其制备方法和用途。ZL01114285.5,聚碳酸酯及其共聚物BTMCPTMC卓仁禧,王旭立,刘立建.,17,In vitro cytotoxicity of PBTMC and PHTMC,COS7 cells.Incubation time:24 h.Temp.:37,o,C,In vitro cytotoxicity of PBTMC,18,BTMC,TMC,Poly(HTMC-co-TMC),Sn(Oct),2,150,o,C,Deprotection,BTMCTMCPoly(HTMC-co-TMC)Sn(Oct,19,Dependence of hydrophilicity on copolymer content,Dependence of hydrophilicity o,20,In vitro degradability of copolymers,PBS pH7.4，37,o,C,0,2,4,6,8,10,12,14,16,18,20,22,24,26,28,0,20,40,60,80,100,P(HTMC-co-TMC)10/90,PTMC,P(HTMC-co-TMC)50/50,Weight Loss(%),Time(day),PHTMC,100%-OH,without-OH,50%-OH,10%-OH,In vitro degradability of copo,21,5-Fu release profiles of copolymers,PBS pH7.4，37,o,C,5-Fu release profiles of copol,22,Zhang Y,Zhuo R X,Synthesis and drug release behavior of poly(trimethylene carbonate)poly(ethylene glycol)poly(trimethylene carbonate)nanoparticles,Biomaterials,(In press),PTET triblock copolymer,纳米药物控制释放,Zhang Y,Zhuo R X,Synthesis a,23,Synthesis and properties of PTET triblock copolymers,Synthesis and properties of PT,24,Particle sizes,drug loading content,loading efficiency,and yield of the PTET polymeric nanoparticles,Polymeric nanoparticles were prepared by the dialysis technique.,Drug:methotrexate(MTX),Particle sizes,drug loading c,25,TEM image of PTET-7 nanoparticles loaded methotrexate(MTX),TEM image of PTET-7 nanopartic,26,Effect of molecular weight of PEG in the copolymers on the MTX release rate,PBS,pH=7.4,37,C.,PEG 6000,PEG 4000,PEG 2000,Effect of molecular weight of,27,Effect of content of PEG block in the copolymers on the MTX release rate,PBS,pH=7.4,37,C.,PEG:6.17 wt%,PEG:24.12 wt%,PEG:28.94 wt%,PEG:54.26 wt%,Effect of content of PEG block,28,温度敏感快速响应PNIPA水凝胶,A 混合溶剂法,B 相分离法,C 使用成孔剂:如PEG,D 形成互穿网络,在温度敏感水凝胶领域发表SCI论文30多篇。,温度敏感快速响应PNIPA水凝胶A 混合溶剂法,29,卓仁禧院士_用于基因和抗癌药物控制释放高分子材料研究(minimizer)课件,30,靶向性抗癌药物,5-Fu抗癌药物,PM,liver-targeting group,肝靶向高分子抗癌药物,靶向性抗癌药物 5-Fu抗癌药物PM liver-targe,31,肝靶向高分子抗癌药物5-Fu-(,99m,Tc DTPA)-PAEA-PM注射后不同时间在大鼠体内各器官的分布,肝靶向高分子抗癌药物5-Fu-(99mTc DTPA)-,32,62.5g/mL,Bel-7204体外凋亡率:51%,135g/mL,Bel-7204体外凋亡率:62%,Control,肝靶向高分子抗癌药物5-Fu-PAEA-PM对人体肝癌细胞(Bel-7204)的体外凋亡实验的荧光照片,62.5g/mL135g/mLControl肝靶向高分子,33,P(TMC-,co,-EEP),Porcine Pancreas Lipase(PPL),Candida Rugosa Lipase(CL),Al(O,i,Pr),3,酶催化开环聚合反应,P(TMC-co-EEP)Porcine Pancreas,34,Ring-opening copolymerization of TMC and EEP catalyzed by,enzymes,Mw/Mn,Mn,Yield(%),Time(d),Molar ratio(TMC:EEP),E(mg/mmol),Lipase,-,500,-,2,10:5,10,-,-,500,-,2,10:1,10,-,1.69,6410,42.1,2,10:5,10,CL,1.83,6440,62.0,2,10:3,10,CL,2.23,7860,75.6,2,10:1,10,CL,2.18,3230,19.8,2,10:5,10,PPL,1.66,8020,60.1,2,10:3,10,PPL,1.71,10250,82.8,2,10:1,10,PPL,2.63,5730,60.2,4,10:2,10,CL,2.97,6010,57.0,3,10:2,10,CL,1.99,6830,50.2,1,10:2,10,CL,2.75,3430,65.2,4,10:2,10,PPL,1.91,3980,72.3,3,10:2,10,PPL,2.47,5970,56.2,1,10:2,10,PPL,1.81,5328,60.2,2,10:2,50,CL,1.87,6500,57.6,2,10:2,25,CL,2.27,6490,58.3,2,10:2,10,CL,2.43,6230,36.5,2,10:2,25,PPL,2.31,9330,68.9,2,10:2,10,PPL,Ring-opening copolymerization,35,Polymerization catalyzed by PPL immobilized on silica microparticles(PISM),Polymerization catalyzed by PP,36,Effect of lipase concentration on Mn and yield,0.0,0.5,1.0,1.5,2.0,2.5,5000,10000,