聚（乙二醇）-癸二酸的可生物降解聚酯作为 β -环糊精-聚轮烷的主链：一种有前途的基因沉默载体。

Biodegradable Polyester of Poly (Ethylene glycol)-sebacic Acid as a Backbone for β -Cyclodextrin-polyrotaxane: A Promising Gene Silencing Vector.

机构信息

Department of Chemical Engineering, Institute of Chemical Technology, Matunga, Mumbai-400019, India.

Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Matunga, Mumbai-400019, India.

出版信息

Curr Gene Ther. 2019;19(4):274-287. doi: 10.2174/1566523219666190808094225.

DOI:10.2174/1566523219666190808094225

PMID:31393245

Abstract

BACKGROUND

Polyrotaxane, a macromolecular interlocked assembly, consisting of cyclodextrin has excellent inclusion capabilities and functionalization capacity, which makes it a versatile material as a vector for gene delivery applications.

OBJECTIVE

A biodegradable linear aliphatic polyester axle composed of Polyethylene Glycol (PEG) and Sebacic Acid (SA) was used to fabricate the β-Cyclodextrin (β-CD) based polyrotaxane as a cationic polymeric vector and evaluated for its potential gene silencing efficiency.

METHODS

The water-soluble aliphatic polyester was synthesized by the solvent esterification process and characterized using viscometry, GPC, FT-IR and 1H NMR spectroscopy. The synthesized polyester was further evaluated for its biodegradability and cellular cytotoxicity. Hence, this water-soluble polyester was used for the step-wise synthesis of polyrotaxane, via threading and blocking reactions. Threading of β-CD over PEG-SA polyester axle was conducted in water, followed by end-capping of polypseudorotaxane using 2,4,6-trinitrobenzenesulfonic acid to yield polyester-based polyrotaxane. For gene delivery application, cationic polyrotaxane (PRTx) was synthesized and evaluated for its gene loading and gene silencing efficiency.

RESULTS AND DISCUSSION

The resulting novel macromolecular assembly was found to be safe for use in biomedical applications. Further, characterization by GPC and H NMR techniques revealed successful formation of PE-β-CD-PRTx with a threading efficiency of 16%. Additionally, the cellular cytotoxicity assay indicated biosafety of the synthesized polyrotaxane, exploring its potential for gene delivery and other biomedical applications. Further, the biological profile of PRTx: siRNA complexes was evaluated by measuring their zeta potential and gene silencing efficiency, which were found to be comparable to Lipofectamine 3000, the commercial transfecting agent.

CONCLUSION

The combinatory effect of various factors such as biodegradability, favourable complexation ability, near zero zeta potentials, good cytotoxicity properties of poly (ethylene glycol)-sebacic acid based β-Cyclodextrin-polyrotaxane makes it a promising gene delivery vector for therapeutic applications.

摘要

背景

聚轮烷是一种由环糊精组成的高分子互锁组装体，具有优异的包合能力和功能化能力，使其成为基因传递应用的多功能载体材料。

目的

用可生物降解的线性脂肪族聚酯轴（由聚乙二醇（PEG）和癸二酸（SA）组成）来制备β-环糊精（β-CD）基聚轮烷作为阳离子聚合物载体，并评估其潜在的基因沉默效率。

方法

通过溶剂酯化法合成水溶性脂肪族聚酯，并通过粘度计、GPC、FT-IR 和 1H NMR 光谱进行表征。进一步评估合成聚酯的生物降解性和细胞毒性。因此，这种水溶性聚酯用于通过穿线和封端反应逐步合成聚轮烷。β-CD 在水中对 PEG-SA 聚酯轴进行穿线，然后用 2,4,6-三硝基苯磺酸对聚假性轮烷进行端封，得到基于聚酯的聚轮烷。用于基因传递应用，合成阳离子聚轮烷（PRTx）并评估其基因负载和基因沉默效率。

结果与讨论

所得到的新型大分子组装体被发现可安全用于生物医学应用。此外，通过 GPC 和 1H NMR 技术进行的表征表明，成功地形成了具有 16%穿线效率的 PE-β-CD-PRTx。此外，细胞毒性测定表明合成的聚轮烷具有生物安全性，探索了其在基因传递和其他生物医学应用中的潜力。此外，通过测量其 Zeta 电位和基因沉默效率来评估 PRTx：siRNA 复合物的生物学特性，发现其与商业转染剂 Lipofectamine 3000 相当。