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阳离子型缩水甘油基聚(氨基酯)-叶酸靶向缀合物的合成与表征及基因传递研究。

Synthesis and characterization of cationic glycidyl-based poly(aminoester)-folic acid targeting conjugates and study on gene delivery.

机构信息

Department of Occupational Safety and Hygiene, Chia-Nan University of Pharmacy and Science, 60 Erh-Jen Rd., Sec. 1, Jen-Te, Taiwan.

出版信息

Molecules. 2012 Jul 30;17(8):9056-69. doi: 10.3390/molecules17089056.

DOI:10.3390/molecules17089056
PMID:22847144
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6268037/
Abstract

A new poly(aminoester) (EPAE-FA) containing folic acid and amino groups in the backbone and side chain was synthesized. EPAE-FA self-assembled readily with the plasmid DNA (pCMV-βgal) in HEPES buffer and was characterized by dynamic light scattering, zeta potential, fluorescence images, and XTT cell viability assays. To evaluate the transfection effect of graft ratio of FA on the EPAE system, EPAE-FA polymers with two different graft ratios (EPAE-FA12k and EPAE-FA14k) were also prepared. This study found that all EPAE-FA polymers were able to bind plasmid DNA and yielded positively charged complexes with nano-sized particles (< 200 nm). To assess the transfection efficiency mediated by EPAE and EPAE-FA polymers, we performed in vitro transfection activity assays using FR-negative (COS-7) and FR-positive (HeLa) cells. The EPAE-FA12k/DNA and EPAE-FA14k/DNA complexes were able to transfect HeLa cell in vitro with higher transfection efficiency than PEI25k/DNA at the similar weight ratio. These results demonstrated that the introduction of FA into EPAE system had a significant effect on transferring ability for FR-positive cells (HeLa). Examination of the cytotoxicity of PEI25k and EPAE-FA system revealed that EPAE-FA system had lower cytotoxicity. In this paper, EPAE-FA seemed to be a novel cationic poly(aminoester) for gene delivery and an interesting candidate for further study.

摘要

一种新的聚（氨基酯）（EPAE-FA），在主链和侧链中含有叶酸和氨基，被合成出来。EPAE-FA 很容易在 HEPES 缓冲液中与质粒 DNA（pCMV-βgal）自组装，并通过动态光散射、Zeta 电位、荧光图像和 XTT 细胞活力测定进行了表征。为了评估叶酸接枝比对 EPAE 系统的转染效果，还制备了两种不同接枝比（EPAE-FA12k 和 EPAE-FA14k）的 EPAE-FA 聚合物。这项研究发现，所有的 EPAE-FA 聚合物都能够结合质粒 DNA，并产生带正电荷的纳米级颗粒（<200nm）复合物。为了评估 EPAE 和 EPAE-FA 聚合物介导的转染效率，我们使用 FR-阴性（COS-7）和 FR-阳性（HeLa）细胞进行了体外转染活性测定。与 PEI25k/DNA 相比，在相似的重量比下，EPAE-FA12k/DNA 和 EPAE-FA14k/DNA 复合物能够更有效地转染 HeLa 细胞。这些结果表明，将 FA 引入 EPAE 系统对 FR-阳性细胞（HeLa）的转染能力有显著影响。PEI25k 和 EPAE-FA 系统细胞毒性的检测表明，EPAE-FA 系统的细胞毒性较低。在本文中，EPAE-FA 似乎是一种新型的阳离子聚（氨基酯），可用于基因传递，是进一步研究的有趣候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/39a61891c5d7/molecules-17-09056-g011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/496bd17b2d04/molecules-17-09056-g012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/46f97c6771ea/molecules-17-09056-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/eb83a169d538/molecules-17-09056-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/56149509d89a/molecules-17-09056-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/7285873aa435/molecules-17-09056-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/a9c9ab8657ce/molecules-17-09056-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/8d35925de83a/molecules-17-09056-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/d98fc47cad7e/molecules-17-09056-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/5df5e2c990c3/molecules-17-09056-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/faa35f929150/molecules-17-09056-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/99068894a0bb/molecules-17-09056-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/39a61891c5d7/molecules-17-09056-g011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/496bd17b2d04/molecules-17-09056-g012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/46f97c6771ea/molecules-17-09056-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/eb83a169d538/molecules-17-09056-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/56149509d89a/molecules-17-09056-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/7285873aa435/molecules-17-09056-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/a9c9ab8657ce/molecules-17-09056-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/8d35925de83a/molecules-17-09056-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/d98fc47cad7e/molecules-17-09056-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/5df5e2c990c3/molecules-17-09056-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/faa35f929150/molecules-17-09056-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/99068894a0bb/molecules-17-09056-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b11/6268037/39a61891c5d7/molecules-17-09056-g011.jpg

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