Semitelechelic HPMA copolymers functionalized with triphenylphosphonium as drug carriers for membrane transduction and mitochondrial localization
Semitelechelic HPMA copolymers functionalized with triphenylphosphonium as drug carriers for membrane transduction and mitochondrial localization
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DOI：
10.1021/bm060336m
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发表时间：
2006-08-14
影响因子：
6.2
通讯作者：
Kopecek, Jindrich
中科院分区：
文献类型：
--
作者：
Callahan, Jon;Kopecek, Jindrich
半遥爪HPMA（N-（2-羟丙基）甲基丙烯酰胺）共聚物具有一个单一的终端亲脂性三苯基膦（TPP）阳离子和荧光标记的合成，以确定如何连接的阳离子影响细胞摄取和细胞内运输。使用分离的小鼠肝线粒体进行的体外线粒体摄取荧光猝灭测定表明，仅较低分子量（< 5 kDa）的BODIPY FL标记的TPP-半遥爪HPMA共聚物表现出显著的细胞器定位或摄取。使用培养的人卵巢癌细胞评价体外细胞摄取和细胞内运输。与研究中使用的所有类型的TPP共聚物一起孵育的细胞似乎仅通过内吞作用内化聚合物，并且所有内化的共聚物在24小时后被限制在溶酶体隔室中。内吞摄取的TPP-HPMA共聚物共轭物是快速的，这表明它们被内化的吸附内吞作用，而不是流体相胞饮。将低分子量（< 5 kDa）和高分子量（> 5 kDa）的半遥爪共聚物显微注射到培养细胞中，表明TPP部分没有显著地将聚合物定位于线粒体。
Semitelechelic HPMA (N-(2-hydroxypropyl)methacrylamide) copolymers possessing a single terminal lipophilic triphenylphosphonium ( TPP) cation and fluorescent labels were synthesized to determine how the attached cation affected cellular uptake and intracellular trafficking. In vitro mitochondrial uptake fluorescence quenching assays using isolated mouse liver mitochondria indicated that only lower molecular weight (< 5 kDa) BODIPY FL-labeled TPP-semitelechelic HPMA copolymers exhibited significant organelle localization or uptake. In vitro cellular uptake and intracellular trafficking was evaluated using cultured human ovarian carcinoma cells. Cells incubated with all types of TPP copolymers used in the study appeared to internalize the polymer by endocytosis only, and all of the internalized copolymer was confined to the lysosomal compartment after 24 h. Endocytotic uptake of the TPP-HPMA copolymer conjugates was rapid, suggesting that they were internalized by adsorptive endocytosis, rather than fluid-phase pinocytosis. Low-molecular weight (< 5 kDa) and high-molecular weight (> 5 kDa) semitelechelic copolymers, microinjected into cultured cells indicated that the TPP moiety did not significantly localize the polymers to mitochondria.