Method for constructing core-shell superstructure dendrimer with amino groups on surface through host and guest self-assembling effect

A technology of host-guest self-assembly and dendrimers, applied in the field of core-shell superstructure dendrimers, to achieve high transfection efficiency, good stability, and good gene transfection effect

Inactive Publication Date: 2017-11-17
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The search results of related literature and patents at home and abroad show that the method of using the host-guest self-assembly in supramolecular chemistry to construct a core-shell superstructure dendrimer for gene transfection has not been reported yet.

Method used

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  • Method for constructing core-shell superstructure dendrimer with amino groups on surface through host and guest self-assembling effect
  • Method for constructing core-shell superstructure dendrimer with amino groups on surface through host and guest self-assembling effect
  • Method for constructing core-shell superstructure dendrimer with amino groups on surface through host and guest self-assembling effect

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Experimental program
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Effect test

Embodiment 1

[0040] Weigh 2.11 mg of 1-adamantaneacetic acid (Ada-COOH) and dissolve it in 5 mL of DMSO. Weigh 20.81mg of EDC and 12.49mg of NHS, dissolve them in 2mL of DMSO respectively, add the EDC solution and DMSO solution to the Ada-COOH solution dropwise, and stir for 3 hours to obtain the activated Ada-COOH solution. Weigh 50 mg of the third-generation polyamidoamine dendrimer (G3), and dissolve it in 5 mL of DMSO. Then the above activated Ada-COOH solution was added dropwise to the G3 solution, and the reaction was stirred at room temperature for 3 days. After the reaction was completed, the reaction solution was transferred to a dialysis bag with a molecular weight cut-off of 1000 Da, dialyzed in phosphate (PBS) buffer solution for 1 day, and then replaced with ultrapure water for dialysis for 2 days. Finally, the product G3-AD (M 1 ), stored at -20°C.

[0041] Weigh 21.82mg β-CD and 31.17mg CDI and dissolve them in 5mL DMSO respectively, and mix and stir for 6h. The fifth-ge...

Embodiment 2

[0044] For the M prepared in Example 1 1 , M 2 , M 3 To characterize. 1 H NMR characterization results as figure 1 As shown in a, the proton peak at chemical shift 1.6-1.9ppm represents the proton peak in the molecular structure of adamantane group, and the proton peak at chemical shift 2.2-3.4ppm represents the proton peak on the amide bond in G3, indicating that Ada- COOH has been successfully modified on the surface of G3. By integrating the proton peaks in these two regions, it was found that 1.3 Ada molecules were modified on each G3; 1 H NMR characterization results as figure 1 As shown in b, the proton peak at chemical shift 2.3-3.2ppm represents the proton peak on the amide bond in G5, and the proton peak at chemical shift 3.5-4.1ppm and 5.1ppm represents the proton peak in the molecular structure of β-CD, It shows that β-CD has been successfully modified on the surface of G5. By integrating the proton peak areas of G5 and β-CD, it was calculated that 8.7 β-CD...

Embodiment 3

[0046] With 3 kinds of materials M prepared in embodiment 1 1 , M 2 , M 3 Vector / pDNA complexes were prepared with different N / P and subjected to gel retardation experiments. Prepare 1% agarose gel containing ethidium bromide (1 mg / mL) in 8 wells, and place at room temperature until the agarose gel is solidified. The N / P ratios are: 0.125:1, 0.25:1, 0.5:1, 1:1, 2:1, 5:1. Add 1 μg pDNA to each well to prepare a vector / pDNA complex and incubate at 37°C for 30min. The pDNA alone without loading was used as a control. After the vector / pDNA complexes were prepared, the corresponding complexes were respectively added to the wells of the agarose gel, and electrophoresed at 80V for 30min. After electrophoresis, the gel was placed in a gel imager to analyze the migration of pDNA in the gel. The result is as Figure 5 shown. All three materials can complex well with pDNA at lower N / P (N / P=2), which hinders the migration of pDNA. You can also see the M 2 and M 3 When N / P = 1, ...

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Abstract

The invention relates to a method for constructing a core-shell superstructure dendrimer with amino groups on the surface through a host and guest self-assembling effect. The method comprises the following steps: synthesizing and characterizing a polyimide-amine dendrimer modified with host and guest molecules; and constructing and characterizing a core-shell superstructure dendrimer nano-compound based on the host and guest self-assembling effect. The core-shell superstructure dendrimer with amino groups on the surface has the advantages of low toxicity, safety, simple transfection conditions, high transfection efficiency, and good potential application prospect in gene therapy.

Description

technical field [0001] The invention belongs to the field of macromolecular nanocarriers, and in particular relates to a method for constructing surface amino core-shell superstructure dendrimers through host-guest self-assembly. Background technique [0002] Gene therapy refers to the introduction of exogenous therapeutic genes into target cells in a certain way to correct or compensate diseases caused by gene defects and abnormalities, so as to achieve the purpose of treating diseases. As a revolutionary treatment, gene therapy has been widely used in the treatment of genetic diseases, tumors and viral diseases, and has achieved certain results. It has become one of the most popular research topics in the field of life sciences and clinical sciences . [0003] The biggest challenge facing gene therapy is to construct safe and efficient gene delivery vectors. Polyamidoamine (PAMAM) dendrimers have been extensively studied as excellent vectors for gene delivery due to thei...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G81/00C08B37/16
CPCC08B37/0015C08G81/00
Inventor 沈明武陈锋孔令丹史向阳
Owner DONGHUA UNIV
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