pH-sensitive polymers that enhance intracellular drug delivery in vivo
pH-sensitive polymers that enhance intracellular drug delivery in vivo
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DOI：
10.1016/s0168-3659(01)00504-1
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发表时间：
2002-01-17
影响因子：
10.8
通讯作者：
Hoffman, AS
中科院分区：
文献类型：
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作者：
Kyriakides, TR;Cheung, CY;Hoffman, AS
对于那些易受溶酶体酶攻击的药物，如DNA、RNA、寡核苷酸、蛋白质和肽，从内体的胞质递送是至关重要的。因此，我们设计了pH敏感的膜破坏性聚合物，以增强药物从酸性内体区室释放到细胞质中。基于溶血研究，我们已经发现一种聚合物，特别是聚（丙基丙烯酸）（PPAA），在pH低于6.5时对膜破坏非常有效。PPAA还显著增强了细胞培养物中lipoplex制剂的体外转染，并且在多达50%血清的存在下也是如此。在这项研究中，我们已经扩展了我们在体外溶血和细胞培养研究，在体内小鼠切除伤口愈合模型。用绿色荧光蛋白（GFP）编码质粒进行的初步研究表明，将含有PPAA的制剂注射到愈合的伤口中导致GFP表达增加。随后，通过给予血管生成抑制剂血小板反应蛋白-2（TSP 2）的正义和反义DNA，我们能够分别改变TSP 2缺失和野生型小鼠的伤口愈合反应。我们的发现表明，当将PPAA加入到lipoplex制剂中时，与对照制剂相比，TSP 2缺失小鼠中TSP 2的表达增强。这些结果表明，PPAA可以增强体内转染，并且抑制TSP 2表达可以导致改善的伤口愈合。这些结果表明，PPAA可以提供药物如DNA的体内功效的显着改善。(C)2002 Elsevier Science B. V.保留所有权利。
Cytosolic delivery from endosomes is critical for those drugs that are susceptible to attack by lysosomal enzymes, such as DNA, RNA, oligonucleotides, proteins and peptides. Therefore, we have designed pH-sensitive, membrane-disruptive polymers to enhance the release of drugs from the acidic endosomal compartment to the cytoplasm. We have found that one polymer in particular, poly(propylacrylic acid) (PPAA), is very effective at membrane disruption at pHs below 6.5, based on hemolysis studies. PPAA also significantly enhances in vitro transfections of lipoplex formulations in cell culture, and does so in the presence of as much as 50% serum. In this study, we have extended our in vitro hemolysis and cell culture studies to an in vivo murine excisional wound healing model. A pilot study with a green fluorescent protein (GFP)-encoding plasmid indicated that injection of formulations containing PPAA into healing wounds resulted in increased GFP expression. Subsequently, by administering sense and antisense DNA for the angiogenesis inhibitor thrombospondin-2 (TSP2), we were able to alter the wound healing response in TSP2-null and wild type mice, respectively. Our findings showed that when PPAA was added to lipoplex formulations, expression of TSP2 was enhanced in TSP2-null mice compared to control formulations. These results show that PPAA can enhance in vivo transfections and that inhibition of TSP2 expression may lead to improved wound healing. These results suggest that PPAA can provide significant improvements in the in vivo efficacy of drugs such as DNA. (C) 2002 Elsevier Science B.V. All rights reserved.