Optimization of PEGylation for cationic triacyl lipid-based siRNA lipoplexes prepared using the modified ethanol injection method for tumor therapy.

IF 3.6 4区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Journal of Liposome Research Pub Date : 2025-05-05 DOI:10.1080/08982104.2025.2498956

Yoshiyuki Hattori, Mizuki Shinkawa, Aya Kurihara, Ryohei Shimizu

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引用次数: 0

Abstract

We previously developed a modified ethanol injection (MEI) method to construct small interfering RNA (siRNA) lipoplexes by mixing a lipid-ethanol solution with an siRNA-containing phosphate-buffered saline solution. Here, we constructed siRNA lipoplexes with 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl)propan-2-yl)amino)-N,N,N-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, and poly(ethylene glycol) (PEG)-lipid using our MEI method. The siRNA lipoplexes were PEGylated with 1, 3, 5, and 10 mol% PEG cholesteryl ether (PEG-Chol), 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (mPEG-DMG), or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(methoxy[polyethylene glycol]) (mPEG-DSPE). PEGylation of siRNA lipoplexes with PEG-Chol did not attenuate the inhibitory effects of Luc and polo-like kinase 1 (PLK1) siRNA lipoplexes on the luciferase (Luc) activity and proliferation of human cervical carcinoma HeLa-Luc, human ovarian cancer SK-OV-3-Luc, and human breast cancer MCF-7-Luc cells stably expressing Luc. In contrast, PEGylated lipoplexes with 10 mol% mPEG-DMG inhibited Luc activity by Luc siRNA but considerably attenuated the PLK1 siRNA-mediated cytotoxic effects. For PEGylated siRNA lipoplexes with mPEG-DSPE, inhibitory effect of Luc siRNA on Luc activity decreased with increasing amounts of PEG modification, and PLK1 siRNA-mediated cytotoxic effects disappeared at more than 3 mol% PEGylation. Erythrocyte aggregation and hemolysis induction by the siRNA lipoplexes were effectively inhibited by 10 mol% PEGylation, irrespective of the PEG-lipid. Compared to those with 1 mol% PEG-Chol, PEGylated siRNA lipoplexes with 10 mol% PEG-Chol potently reduced siRNA accumulation in mouse lungs post-intravenous administration. Overall, TC-1-12-based siRNA lipoplexes with 10 mol% PEG-Chol exerted PLK1 siRNA-mediated cytotoxic effects, without inducing hemolysis and erythrocyte aggregation.

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用改良的乙醇注射法制备的阳离子三酰基脂基siRNA脂质体聚乙二醇化的优化。

我们之前开发了一种改进的乙醇注射（MEI）方法，通过将脂质乙醇溶液与含有siRNA的磷酸盐缓冲盐水溶液混合来构建小干扰RNA （siRNA）脂质体。在这里，我们用我们的MEI方法构建了11-（（1,3-二（十二烷基氧基）-2-（十二烷基氧基）甲基）丙烷-2-基）氨基-N，N，N-三甲基-11-氧十二烷基-1-溴化胺（TC-1-12）， 1,2-二油基-sn-甘油-3-磷酸乙醇胺和聚乙二醇(PEG)-脂质siRNA脂质体。siRNA脂质体分别用1,3,5和10 mol%的PEG胆固醇醚（PEG- chol）、1,2-二myristoyl- racc -glycero-3-甲氧基聚乙二醇（mPEG-DMG）或1,2-二硬脂酰-sn-glycero-3- phospho乙醇胺- n-（甲氧基聚乙二醇）（mPEG-DSPE）聚乙二醇化。用PEG-Chol修饰siRNA脂质团，不会减弱Luc和polo样激酶1 (PLK1) siRNA脂质团对稳定表达Luc的人宫颈癌HeLa-Luc、人卵巢癌SK-OV-3-Luc和人乳腺癌MCF-7-Luc细胞荧光素酶（Luc）活性和增殖的抑制作用。相比之下，含有10 mol% mPEG-DMG的聚乙二醇化脂质体通过Luc siRNA抑制Luc活性，但显著减弱PLK1 siRNA介导的细胞毒性作用。对于mPEG-DSPE修饰的聚乙二醇化siRNA脂质体，Luc siRNA对Luc活性的抑制作用随着PEG修饰量的增加而降低，PLK1 siRNA介导的细胞毒性作用在超过3mol %的PEG修饰时消失。无论peg脂质如何，10 mol%的PEGylation都能有效抑制siRNA脂质团的红细胞聚集和溶血诱导。与含有1mol % PEG-Chol的小鼠相比，含有10mol % PEG-Chol的聚乙二醇化siRNA脂质体在静脉给药后可有效减少小鼠肺部siRNA的积累。总体而言，含有10 mol% PEG-Chol的tc -1-12基siRNA脂质体发挥PLK1 siRNA介导的细胞毒性作用，不诱导溶血和红细胞聚集。

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来源期刊

Journal of Liposome Research 生物-生化与分子生物学

CiteScore

10.50

自引率

2.30%

发文量

审稿时长

3 months

期刊介绍： The Journal of Liposome Research aims to publish original, high-quality, peer-reviewed research on the topic of liposomes and related systems, lipid-based delivery systems, lipid biology, and both synthetic and physical lipid chemistry. Reviews and commentaries or editorials are generally solicited and are editorially reviewed. The Journal also publishes abstracts and conference proceedings including those from the International Liposome Society. The scope of the Journal includes: Formulation and characterisation of systems Formulation engineering of systems Synthetic and physical lipid chemistry Lipid Biology Biomembranes Vaccines Emerging technologies and systems related to liposomes and vesicle type systems Developmental methodologies and new analytical techniques pertaining to the general area Pharmacokinetics, pharmacodynamics and biodistribution of systems Clinical applications. The Journal also publishes Special Issues focusing on particular topics and themes within the general scope of the Journal.