Development of a novel endosomolytic diblock copolymer for siRNA delivery.
Development of a novel endosomolytic diblock copolymer for siRNA delivery.
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DOI：
10.1016/j.jconrel.2008.10.004
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发表时间：
2009-02-10
期刊：
影响因子：
--
通讯作者：
Stayton PS
中科院分区：
文献类型：
--
作者：
Convertine AJ;Benoit DS;Duvall CL;Hoffman AS;Stayton PS
小干扰RNA(SiRNA)的基因敲除活性使其成为靶向验证工具和多种疾病的潜在治疗药物。然而，事实证明，这些双链RNA大分子的输送是具有挑战性的，在许多情况下，是它们部署的障碍。在这里，我们报告了一种新的两嵌段共聚物家族的开发，该家族旨在增强siRNA的全身和细胞内递送。采用可控可逆加成断裂链转移聚合(RAFT)方法合成了两嵌段共聚物，其组成包括一个带正电的甲基丙烯酸二甲氨基乙酯(DMAEMA)嵌段和第二个由DMAEMA和丙基丙烯酸(PAA)组成的大致等摩尔比的内质体释放嵌段，以及甲酸丁酯(BMA)。表征了一系列相关的双嵌段组成，阳离子嵌段保持不变，DMAEMA和PAA与BMA的比例不同。这些载体在内体pH范围内变得急剧溶血，溶血活性随着第二块中BMA所占比例的系统性增加而增加。两嵌段共聚物将siRNA凝聚成具有轻微正Zeta电位的80-250 nm颗粒。SiRNA介导的模型蛋白，即甘油醛3-磷酸脱氢酶(GAPDH)在HeLa细胞中的敲除通常遵循溶血活性趋势，最疏水的第二块(最高的BMA含量)表现出最好的敲除。这种pH响应型载体被设计用来调节内体释放，显示出在细胞内传递siRNA的重要前景。
The gene knockdown activity of small interfering RNA (siRNA) has led to their use as target validation tools and as potential therapeutics for a variety of diseases. The delivery of these double-stranded RNA macromolecules has proven to be challenging, however, and in many cases, is a barrier to their deployment. Here we report the development of a new diblock copolymer family that was designed to enhance the systemic and intracellular delivery of siRNA. These diblock copolymers were synthesized using the controlled reversible addition fragmentation chain transfer polymerization (RAFT) method and are composed of a positively-charged block of dimethylaminoethyl methacrylate (DMAEMA) to mediate siRNA condensation, and a second endosomal-releasing block composed of DMAEMA and propylacrylic acid (PAA) in roughly equimolar ratios, together with butyl methacylate (BMA). A related series of diblock compositions were characterized, with the cationic block kept constant, and with the ratio of DMAEMA and PAA to BMA varied. These carriers became sharply hemolytic at endosomal pH regimes, with increasing hemolytic activity seen as the percentage of BMA in the second block was systematically increased. The diblock copolymers condensed siRNA into 80–250 nm particles with slightly positive Zeta potentials. SiRNA-mediated knockdown of a model protein, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), in HeLa cells generally followed the hemolytic activity trends, with the most hydrophobic second block (highest BMA content) exhibiting the best knockdown. This pH-responsive carrier designed to mediate endosomal release shows significant promise for the intracellular delivery of siRNA.
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