高分子基因治疗载

1、 高分子基因治疗载体高分子基因治疗载体病毒载体和非病毒载体的比较病毒载体和非病毒载体的比较效率低效率低安全安全容易制备，储存和质量控制容易制备，储存和质量控制传递的传递的DNADNA大小从大小从2020个碱基对个碱基对到到几十万个碱基对几十万个碱基对非病毒非病毒载体载体安全隐患安全隐患DNADNA大小受限制大小受限制免疫原性免疫原性高效高效病毒病毒载体载体缺点缺点优点优点载体载体 非病毒载体非病毒载体 阳离子脂质体阳离子脂质体 高分子高分子: : 阳离子高分子阳离子高分子 水溶性非离子高分子水溶性非离子高分子 高分子基质高分子基质 常用的聚阳离子基因载体常用的聚阳离子基因载体OOH2NnH2N

2、NHNNHNHNH2NHNNNHHNNHPoly-(4-amino-butyl)-L-glycolic acidS W Kim, Utah UnivPoly-L-lysinePolyethylenimines (PEI)Behr J P, FranceNHOH2NnDendrimerSzoka UCSF & Baker J Univ MichNHHNOOHN(CH2)4NH2OHNNxyOPOOONH3ClnPPEOPOONnH2NNH2PPA-SPPolyphosphoester and PolyphosphoamideLeong K W, Johns Hopking UnivLanger

3、R, MIT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HNHNNHHNHOOHHOHOOHOHOHOHHOHOCOOMeCOOMeMeOOCNHNONNHHNOONNHHNOONNHHNOONNHHNOONH2NH2NNHHNOONH2NH2NNHHNOONH2NH2NNHNHOONH2NH2HNONNHHNOONNHHNOONNHHNOONNHHNOOH2NNH2NNHHNOOH2NNH2NNHHNOOH2NNH2NNHHNOONH2NH2NHOONNHONHNNHOONHNNHOONHNNHOONHNNHOONH2NH2NHNNHOONH2NH2N

4、HNNHOONH2H2NNHNNHOONH2H2NHNONNHNHOONNHNHOONNHNHOONNHHNOONH2NH2NNHHNOONH2NH2NNHNHOONH2NH2NHNNHOONH2NH2HNOOCNNHONHNNHOONHNNHOONHNNHOONHNNHOOH2NH2NNHNNHOOH2NH2NNHNNHOOH2NH2NNHNHNOOH2NH2NNHONHNNHOONHNNHOONHNNHOONHNNHOONH2H2NNHNNHOONH2H2NNHNNHOONH2H2NNHNNHOONH2H2NHNOONHNONNHHNOONNHHNOONNHHNOONNHHNOOH2NNH

5、2NNHHNOOH2NNH2NNHHNOOH2NNH2NNHHNOOH2NNH2NHONHNHNOONHNHNOONHNHNOONHNNHOOH2NH2NNHNNHOOH2NH2NNHNHNOOH2NH2NNNHHNOOH2NH2NNHOODendrimer GT51.00E+031.00E+041.00E+051.00E+061.00E+071.00E+081.00E+091.00E+10Naked DNATransfastPEIPLLGT3GT4GT5RLU/mg proteinTransfection of HEK 293 cells with pRE Luc/dendrimer GT

6、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，但是比相当，但是比PEI的毒性低得多的毒性低得多020406080100120050100150200Dendrimer concentration (ug/mL)Percent relative viabilityGT3GT4GT5PLLPEICytotoxicity of dendrimer GT3GT5 measured by the MTT assay. PEI and PLL were used as controls 肝细胞靶向基因载体肝细胞靶向基因载体 O

7、OOHOOHOHOHOHOHHOHOPAMAM dendrimer+NaBH3(CN)Dendrimer-GalGT5-Gal 5%GT5-Gal 10%GT5-Gal 20%1.0E+041.0E+051.0E+061.0E+071.0E+081.0E+090246810N/PRLU/mg proteinGT5GT5-Gal 5%Transfection of HepG2 cells with pRE Luc1.00E+031.00E+041.00E+051.00E+061.00E+071.00E+081.00E+090246810N/PRLU/mg proteinGT5GT5-Gal 5%Transfection of NIH 3T3 cells with pRE Luc阳离子聚磷酸酯基因载体阳离子聚磷酸酯基因载体OPOOO(CH2)nNRH2ClnPPE-HA: R=H, n=6PPE-MEA: R=Me, n=2PPA-DPAGal-PPAOCH2CHCH3OPONH2NNHOCH2CHCH3OPONH2NNH2xyOOHOHHOOHNHOCH2COPEGSOOGal-PEG-PPAliverspleenlungkidneyheart103105104102RLU/mg protei