Application of hydroxyl-containing crosslinked polymer guanidinated product in gene transfer

Abstract

The invention discloses a preparation method and an application of a hydroxyl-containing crosslinked polymer and guanidinated product thereof used for delivering nucleic acid and other bioactive agents. The method comprises the specific steps that: linear or branched polyethyleneimine with a molecular weight of 100-30000Da or various multi-amine compounds are subjected to a compounding reaction with one or more crosslinking agents with a molecular weight of 100-5000Da and containing two or more epoxy groups, such that the hydroxyl-containing crosslinked polymer is prepared. The invention also provides a synthesizing method of a novel guanidination reagent 3-guanidino propanol acrylate or 5-guanidino amyl acrylate. With the guanidination reagent, the hydroxyl-containing crosslinked polymer or other polymers can be guanidinated, such that gene transfection efficiency thereof can be substantially improved, and cytotoxicity can be reduced. The excellent-performance polymers screened by the invention have gene transfection efficiencies in various cell lines such as A549, B16F10, 3T3, and U87 substantially higher than conventional commercialized gene transfection reagents. The polymers are low-toxic high-efficiency non-virus gene transfection vectors. With the vectors provided by the invention, in-vitro high-efficiency and low-toxicity transfection of various cells can be realized, and in-vivo high-efficiency and low-toxicity local administration can be realized. The application has wide application prospect.

Description

technical field

[0001] The invention relates to a method for crosslinking polyethyleneimine and its analogues with two or more epoxy group crosslinking agents to form hydroxyl-containing crosslinked polymers. Simultaneously also relate to the guanidinating reagent with certain chain length, i.e. 3-guanidinopropanol acrylate, the synthetic method of 5-guanidinopentyl alcohol acrylate etc. A method for the preparation of guanidinated polymers by chemical reaction. technical background

[0002] It was discovered earlier that the human immunodeficiency virus (HIV) type 1 transactivator (TAT) has the ability to penetrate various types of cell membranes, which can greatly increase the transcription and replication levels of the HIV genome. According to the function of TAT, a class of small molecular polypeptides with the ability to penetrate cell membranes has been discovered, called cell penetrating peptides (Cell Penetrating Peptides, CPPs), whose length generally does not exce...

Images