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Endogenous hyperbranched poly-spermine kation genetic vector as well as preparation method and application thereof

A gene carrier, endogenous technology, applied in the field of biomedical engineering materials, to achieve the effects of good biocompatibility, high product yield and simple material composition

Active Publication Date: 2018-09-14
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, combining hyperbranched structures with endogenous monomers to construct endogenous hyperbranched cationic gene carriers and their applications have not been reported

Method used

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  • Endogenous hyperbranched poly-spermine kation genetic vector as well as preparation method and application thereof
  • Endogenous hyperbranched poly-spermine kation genetic vector as well as preparation method and application thereof
  • Endogenous hyperbranched poly-spermine kation genetic vector as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: the preparation of hyperbranched polyspermine cation

[0051] (1) Weigh 2 mg of spermine and dissolve it in 5 ml of anhydrous chloroform, and use 2-acetyl-5,5-dimethyl-1,3-cyclohexylamine diketone (Dde-OH) under anhydrous and anaerobic conditions (5.4 mg) protected its two primary amine groups. The reaction was carried out for 24 hours at a rotation speed of 600 rpm and a temperature of 25°C. After the reaction is completed, the protected spermine monomer is obtained by removing impurities; wherein, the molar ratio of spermine to Dde-OH is 1:3.

[0052] (2) Weigh (5mg) the protected spermine monomer and dissolve it in 10mL of water, add trimethylolpropane triacrylate (0.96mg), stir and react for 6 days under the condition of rotating speed of 600rpm and temperature of 65°C . After the reaction was completed, the polymer was precipitated by adding anhydrous ether; to the precipitated polymer, 10ml of deprotection solution (2.01g of imidazole, 8mL of N-me...

Embodiment 2

[0053] Embodiment 2: the proton spectrum NMR characterization of hyperbranched polyspermine cation

[0054] The hyperbranched polyspermine cation obtained in Example 1 was dissolved in deuterated water for hydrogen spectrum nuclear magnetic characterization. Such as figure 2 , the chemical shifts of the hydrogen atoms in the hyperbranched polyspermine cation have been annotated, figure 2 The result proves that this step reaction successfully synthesized the hyperbranched polyspermine cation.

Embodiment 3

[0055] Embodiment 3: the preparation of hyperbranched polyspermine cation and pDNA nanocomposite

[0056] (1) take by weighing the hyperbranched polyspermine cation that makes among the embodiment 1 and be dissolved in water, obtain hyperbranched polyspermine cation solution;

[0057] (2) pDNA (DNA, Suzhou Gemma Pharmaceutical Technology Co., Ltd.) is configured as a 1mg / ml stock solution;

[0058] (3) Preparation of hyperbranched polyspermine cation and pDNA nanocomposite: according to a series of mass ratios (1, 5, 10, 15, 20, 30) set, dilute the hyperbranched polyspermine cation solution to 1 mg / ml , add 1, 5, 10, 15, 20, 30ml of polyspermine cation solution to each test tube, then quickly add 1ml of pDNA stock solution, and incubate at 25°C for 30 minutes to obtain a series of nanoparticles with different surface charge properties. Complex.

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Abstract

The invention discloses an endogenous hyperbranched poly-spermine kation genetic vector as well as a preparation method and application thereof. The preparation method of the genetic vector comprisesthe following steps that 1, an endogenous monomer is dissolved in organic solvent, under the water-free and oxygen-free conditions, a protective agent is added, stirring reaction is carried out at 20-30 DEG C, and a protected endogenous monomer I is obtained; 2, the endogenous monomer I is dissolved in water, then a biodegradable bridging agent is added, addition reaction is carried out at 60-80 DEG C, then diethyl ether is added to make a product precipitate, and a polymer II is obtained; 3, a deprotection agent is added into the polymer II for reaction, filtering and freeze drying are conducted, and the endogenous hyperbranched polyamine kation genetic vector is obtained. The genetic vector obtained through the method has both biodegradability and non-toxic metabolism and can agglomerateand convey nucleic acid medicines, nano-composite particles used for curing DME can be obtained by compounding the genetic vector and the nucleic acid medicines, and thus safe efficient gene therapyis achieved.

Description

technical field [0001] The invention belongs to the field of biomedical engineering materials, in particular to an endogenous hyperbranched polyspermine cationic gene carrier and its preparation method and application. Background technique [0002] Among all complications of diabetes, the average prevalence of diabetic retinopathy is as high as 50%. Diabetic retinopathy occurs when diabetes affects the retina. Diabetic macular edema (DME) is the primary cause of vision loss in diabetic patients. In the late course of the disease, total retinal detachment can lead to complete blindness. Gene therapy has become a relatively potential treatment strategy for DME at present, especially ribonucleic acid interference technology, which not only provides a new method for studying the gene expression and regulation functions of organisms, but also opens up a new way for the research, prevention and treatment of DME . In gene therapy, RNAi (RNA interference) is a powerful tool for si...

Claims

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Application Information

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IPC IPC(8): C12N15/87C08G83/00A61K48/00
CPCA61K48/0041C08G83/005C12N15/87
Inventor 薛巍纪鑫马栋郭会龙
Owner JINAN UNIVERSITY