Hydroxyl stereochemistry and amine number within poly(glycoamidoamine)s affect intracellular DNA delivery
Hydroxyl stereochemistry and amine number within poly(glycoamidoamine)s affect intracellular DNA delivery
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DOI：
10.1021/ja0436446
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发表时间：
2005-03-09
影响因子：
15
通讯作者：
Reineke, TM
中科院分区：
文献类型：
--
作者：
Liu, YM;Reineke, TM
核酸药物在治疗多种恶性肿瘤方面具有巨大的潜力，但由于缺乏有效、无毒的载体，限制了其应用。本文通过酯化D-葡糖二酸（D）、内消旋半乳糖二酸二甲酯（G）和（D）-甘露糖-1，4：6，3-二内酯（M）与二亚乙基三胺（1）、三亚乙基四胺（2）、四亚乙基五胺（3）和五亚乙基六胺（4）的缩聚反应合成了一种新的聚（葡糖酰胺胺）（D1 -D4、G1 -G4和M1 -M4）库。碳水化合物羟基的立体化学和胺单元的数量已经系统地改变，以努力检查聚合物化学如何影响质粒DNA（pDNA）结合亲和力、pDNA压缩成纳米颗粒（聚合复合物）、材料细胞毒性和核酸递送的功效。发现在碳水化合物之间具有四个仲胺（D4、G4和M4）的聚合物具有最高的pDNA结合亲和力，并且半乳糖二酸酯聚合物通常产生最小的聚合复合物。在BHK-21、HeLa和HepG 2细胞中使用含有萤火虫荧光素酶或β-半乳糖苷酶报告基因的pDNA的递送研究证明，所有聚（葡糖胺）递送pDNA而没有细胞毒性。聚合物D4、G4和M4显示出最高的递送效率，其中发现G4是特别有效的递送载体。肝素竞争试验表明，这可能是G4显示的pDNA结合亲和力高于D4和M4的结果。由具有较弱pDNA亲和力的聚合物形成的聚合复合物可能由于与带负电荷的糖胺聚糖的相互作用而在细胞表面解离，这将导致包合的聚合复合物的数量减少。
Nucleic acid drugs have great potential to treat many devastating aliments, but their application has been hindered by the lack of efficacious and nontoxic delivery vehicles. Here, a new library of poly(glycoamidoamine)s (D1 -D4, G1 -G4, and M1 -M4) has been synthesized by polycondensation of esterified D-glucaric acid (D), dimethyl-meso-galactarate (G), and (D)-mannaro-1,4:6,3-dilactone (M) with diethylenetriamine (1), triethylenetetramine (2), tetraethylenepentamine (3), and pentaethylenehexamine (4). The stereochemistry of the carbohydrate hydroxyl groups and the number of amine units have been systematically changed in an effort to examine how the polymer chemistry affects the plasmid DNA (pDNA) binding affinity, the compaction of pDNA into nanoparticles (polyplexes), the material cytotoxicity, and the efficacy of nucleic acid delivery. The polymers with four secondary amines (D4, G4, and M4) between the carbohydrates were found to have the highest pDNA binding affinity and the galactarate polymers generally yielded the smallest polyplexes. Delivery studies with pDNA containing the firefly luciferase or beta-galactosidase reporter genes in BHK-21, HeLa, and HepG2 cells demonstrated that all of the poly(glycoamicloamine)s deliver pDNA without cytotoxicity. Polymers D4, G4, and M4 displayed the highest delivery efficiency, where G4 was found to be a particularly effective delivery vehicle. Heparin competition assays indicated that this may be a result of the higher pDNA binding affinity displayed by G4 as compared to D4 and M4. Polyplexes formed by polymers with weaker pDNA affinities may dissociate at the cell surface due to interactions with negatively charged glycosaminoglycans, which would cause a decrease in the number of polyplexes that are enclocytosed.