Institute of Organic Chemistry and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Lessingstrasse 8, Jena, 07743, Germany.
Pharmaceutical Technology and Biopharmacy, Institute of Pharmacy, Friedrich Schiller University Jena, Lessingstrasse 8, Jena, 07743, Germany.
Macromol Biosci. 2021 Oct;21(10):e2100146. doi: 10.1002/mabi.202100146. Epub 2021 Jul 26.
This study describes the first example for shielding of a high performing terpolymer that consists of N-(2-hydroxypropyl)methacrylamide (HPMA), N-(3-guanidinopropyl)methacrylamide (GPMA), and N-(2-indolethyl)methacrylamide monomers (IEMA) by block copolymerization of a polyethylene glycol derivative - poly(nona(ethylene glycol)methyl ether methacrylate) (P(MEO MA)) via reversible addition-fragmentation chain transfer (RAFT) polymerization. The molecular weight of P(MEO MA) is varied from 3 to 40 kg mol while the comonomer content of HPMA, GPMA, and IEMA is kept comparable. The influence of P(MEO MA) block with various molecular weights is investigated over cytotoxicity, plasmid DNA (pDNA) binding, and transfection efficiency of the resulting polyplexes. Overall, the increase in molecular weight of P(MEO MA) block demonstrates excellent biocompatibility with higher cell viability in L-929 cells and an efficient binding to pDNA at N/P ratio of 2. The significant transfection efficiency in CHO-K1 cells at N/P ratio 20 is obtained for block copolymers with molecular weight of P(MEO MA) up to 10 kg mol . Moreover, a fluorescently labeled analogue of P(MEO MA), bearing perylene monoimide methacrylamide (PMIM), is introduced as a comonomer in RAFT polymerization. Polyplexes consisting of labeled block copolymer with 20 kg mol of P(MEO MA) and pDNA are incubated in Hela cells and investigated through structured illumination microscopy (SIM).
本研究描述了首例通过聚乙二醇衍生物 - 聚（壬（乙二醇）甲基醚甲基丙烯酸酯）（P（MEO MA））的可逆加成-断裂链转移（RAFT）聚合对由 N-（2-羟丙基）甲基丙烯酰胺（HPMA）、N-（3-胍丙基）甲基丙烯酰胺（GPMA）和 N-（2-吲哚乙基）甲基丙烯酰胺单体（IEMA）组成的高性能三元共聚物进行屏蔽的示例。P（MEO MA）的分子量从 3 至 40 kg mol 变化，而 HPMA、GPMA 和 IEMA 的共聚单体含量保持可比。研究了不同分子量的 P（MEO MA）嵌段对细胞毒性、质粒 DNA（pDNA）结合和所得聚合物的转染效率的影响。总体而言，P（MEO MA）嵌段的分子量增加表现出与 L-929 细胞更高的细胞活力和在 N/P 比为 2 时与 pDNA 的有效结合的优异生物相容性。在 N/P 比为 20 时，CHO-K1 细胞中获得了显著的转染效率，分子量为 10 kg mol 的 P（MEO MA）的嵌段共聚物。此外，将带有芘单酰亚胺甲基丙烯酰胺（PMIM）的荧光标记的 P（MEO MA）类似物作为共聚单体引入 RAFT 聚合中。将包含 20 kg mol 的 P（MEO MA）和 pDNA 的标记嵌段共聚物的聚合物与 pDNA 一起孵育在 Hela 细胞中，并通过结构光照明显微镜（SIM）进行研究。
