Neutral polymeric micelles for RNA delivery.
Neutral polymeric micelles for RNA delivery.
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DOI：
10.1021/bc300486k
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发表时间：
2013-03-20
影响因子：
4.7
通讯作者：
Stayton, Patrick S.
中科院分区：
文献类型：
--
作者：
Lundy, Brittany B.;Convertine, Anthony;Miteva, Martina;Stayton, Patrick S.
RNA干扰(RNAi)药物具有巨大的治疗潜力，但仍需要具有适当疗效和毒性的递送系统。在这里，我们描述了一种基于可共轭双嵌段聚合物胶束的中性、两性聚合物递送系统。这种两嵌段共聚物包含一个亲水性的聚[N-(2-羟丙基)甲基丙烯酰胺-N-(2-(吡啶-2-基二硫基)乙基)甲基丙烯酰胺](聚[HPMA-co-PDSMA])链段，以提高水溶液的稳定性和促进硫醇-二硫化物交换反应，以及由丙基丙烯酸(PAA)、甲基丙烯酸二甲氨基乙酯(DMAEMA)和甲基丙烯酸丁酯(BMA)组成的第二个两性嵌段。采用可逆加成-断裂链转移(RAFT)聚合方法合成了总相对分子质量为22,000g/m o l，pDI值为1.88的poly[(HPMA-co-PDSMA)-b-(PAA-co-DMAEMA-co-BMA)]。动态光散射和荧光测量表明，两嵌段共聚物在水溶液中自组装形成聚合物胶束，其流体动力学半径和临界胶束浓度分别为25 nm和25μg/mL。红细胞溶血实验表明，中性亲水胶束在膜的pH值范围内具有较强的膜破坏活性。针对甘油醛3-磷酸脱氢酶(GAPDH)的硫代化siRNA通过与胶束冠中吡啶二硫基团的硫醇交换反应直接偶联到聚合物胶束上。在孵育48h后，当siRNA与聚合物的摩尔比为1：10时，HeLa细胞的沉默活性最高。在此条件下，48h的信使核糖核酸和蛋白质的抑制率分别为90%和65%，毒性很小。相反，在这些条件下，缺乏pH响应性内切酶片段的聚合物胶束表现出可以忽略不计的mRNA和蛋白质敲除。中性siRNA结合物强大的mRNA敲除能力和良好的生物相容性表明，这种载体设计用于输送治疗性siRNA药物具有潜在的实用价值。
RNA interference (RNAi) drugs have significant therapeutic potential but delivery systems with appropriate efficacy and toxicity profiles are still needed. Here, we describe a neutral, ampholytic polymeric delivery system based on conjugatable diblock polymer micelles. The diblock copolymer contains a hydrophilic poly[N-(2-hydroxypropyl) methacrylamide-co-N-(2-(pyridin-2- yldisulfanyl)ethyl)methacrylamide) (poly[HPMA-co-PDSMA]) segment to promote aqueous stability and facilitate thiol-disulfide exchange reactions, and a second ampholytic block composed of propyl acrylic acid (PAA), dimethylaminoethyl methacrylate (DMAEMA), and butyl methacrylate (BMA). The poly[(HPMA-co-PDSMA)-b-(PAA-co-DMAEMA-co-BMA)] was synthesized using Reversible Addition-Fragmentation chain Transfer (RAFT) polymerization with an overall molecular weight of 22,000 g/mol and a PDI of 1.88. Dynamic light scattering and fluorescence measurements indicated that the diblock copolymers self-assemble under aqueous conditions to form polymeric micelles with a hydrodynamic radius and critical micelle concentration of 25 nm and 25 μg/mL respectively. Red blood cell hemolysis experiments show that the neutral hydrophilic micelles have potent membrane destabilizing activity at endosomal pH values. Thiolated siRNA targeting glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was directly conjugated to the polymeric micelles via thiol exchange reactions with the pyridal disulfide groups present in the micelle corona. Maximum silencing activity in HeLa cells was observed at a 1:10 molar ratio of siRNA to polymer following a 48 h incubation period. Under these conditions 90 % mRNA knockdown and 65 % and protein knockdown of at 48 h was achieved with negligible toxicity. In contrast the polymeric micelles lacking a pH-responsive endosomalytic segment demonstrated negligible mRNA and protein knockdown under these conditions. The potent mRNA knockdown and excellent biocompatibility of the neutral siRNA conjugates demonstrate the potential utility if this carrier design for delivering therapeutic siRNA drugs.
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DOI：
10.1016/j.jconrel.2011.06.013
发表时间：
2011-10-30
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Crownover E;Duvall CL;Convertine A;Hoffman AS;Stayton PS
通讯作者：
Stayton PS
DOI：
10.1016/j.jconrel.2008.10.004
发表时间：
2009-02-10
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Convertine AJ;Benoit DS;Duvall CL;Hoffman AS;Stayton PS
通讯作者：
Stayton PS
影响因子：
12.4
作者：
Howard, Kenneth A.;Rahbek, Ulrik L.;Kjems, Jorgen
通讯作者：
Kjems, Jorgen
影响因子：
4.5
作者：
Khan, A;Benboubetra, M;Akhtar, S
通讯作者：
Akhtar, S
影响因子：
--
作者：
Chiu, YL;Ali, A;Rana, TM
通讯作者：
Rana, TM