Quaternary complexes modified from pDNA and poly-l-lysine complexes to enhance pH-buffering effect and suppress cytotoxicity.
Quaternary complexes modified from pDNA and poly-l-lysine complexes to enhance pH-buffering effect and suppress cytotoxicity.
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DOI：
10.1002/jps.24364
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发表时间：
2015-04
影响因子：
3.8
通讯作者：
Yukinobu Kodama;Yuiko Yatsugi;T. Kitahara;Tomoaki Kurosaki;Kanoko Egashira;M. Nakashima;Takahiro Muro;Hiroo Nakagawa;N. Higuchi;Tadahiro Nakamura;H. Sasaki
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文献类型：
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作者：
Yukinobu Kodama;Yuiko Yatsugi;T. Kitahara;Tomoaki Kurosaki;Kanoko Egashira;M. Nakashima;Takahiro Muro;Hiroo Nakagawa;N. Higuchi;Tadahiro Nakamura;H. Sasaki
通过添加聚组氨酸（PLH）和γ-聚谷氨酸（γ-PGA），将pDNA与聚赖氨酸（PLL）复合，以增强其pH缓冲作用并抑制细胞毒性。pDNA与PLL或/和PLH的二元和三元复合物显示具有阳离子表面电荷的约52-76 nm的粒径。三元复合物显示出比具有PLL的二元复合物高得多的基因表达。PLL和PLH的混合溶液比PLL溶液具有更高的缓冲能力。三元复合物的高基因表达被巴弗洛霉素A1降低。这些结果表明，PLH的PLL复合物的添加促进内体逃逸，通过增强pH缓冲效应。由于配合物表面带阳离子电荷，因此具有细胞毒性和血凝作用。因此，我们通过添加阴离子γ-PGA开发了四元复合物，据报道，这可以降低阳离子复合物的毒性。事实上，四元配合物没有显示出细胞毒性和血液凝集。此外，四元复合物表现出更高的基因表达比三元复合物，无论其阴离子表面电荷。四元复合物显示出选择性的高基因表达后，他们的静脉内给药的脾脏。因此，我们成功地开发了具有高基因表达和无毒性的四元配合物。
We developed a modified complex of pDNA and poly-l-lysine (PLL) by the addition of poly-l-histidine (PLH) and γ-polyglutamic acid (γ-PGA) to enhance its pH-buffering effect and suppress cytotoxicity. The binary and ternary complexes of pDNA with PLL or/and PLH showed particle sizes of approximately 52-76 nm with cationic surface charge. The ternary complexes showed much higher gene expression than the binary complexes with PLL. The mixed solution of PLL and PLH showed higher buffering capacity than PLL solution. The high gene expression of ternary complexes was reduced by bafilomycin A1 . These results indicated the addition of PLH to PLL complexes promoted endosomal escape by enhancing the pH-buffering effect. The binary and ternary complexes showed cytotoxicity and blood agglutination because of their cationic surface charge. We therefore developed quaternary complexes by the addition of anionic γ-PGA, which was reported to decrease the toxicity of cationic complexes. In fact, quaternary complexes showed no cytotoxicity and blood agglutination. Also, quaternary complexes showed higher gene expression than ternary complexes regardless of their anionic surface charge. Quaternary complexes showed selectively high gene expression in the spleen after their intravenous administration. Thus, we successfully developed the quaternary complexes with high gene expression and no toxicity.