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Application method for gene transfection by using polyamide-amine dendritic polymer and gold nano-particle compound as non-viral vector

A technology of amine dendrimer and gold nanoparticles, which is applied in the application field of gene transfection of the complex of polyamidoamine dendrimer and gold nanoparticles as a non-viral vector, and can solve the problem of immune safety detection. See reports and other issues to achieve good gene transfection effect, low immunogenicity and good biocompatibility

Inactive Publication Date: 2017-09-08
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Retrieval of related literature and patents at home and abroad showed that polyamide-amine dendrimers were co-modified with polyamide-amine dendrimers and gold nanoparticles were used as gene carriers for gene transfection. Post-immune safety testing has not been reported

Method used

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  • Application method for gene transfection by using polyamide-amine dendritic polymer and gold nano-particle compound as non-viral vector
  • Application method for gene transfection by using polyamide-amine dendritic polymer and gold nano-particle compound as non-viral vector
  • Application method for gene transfection by using polyamide-amine dendritic polymer and gold nano-particle compound as non-viral vector

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] 20.25 mg of mPEG (molecular weight: 2000) was weighed and dissolved in DMSO, 9.59 mg of EDC and mPEG solution were evenly mixed in a molar ratio of 10:1, and then 5.25 mg of NHS was added to react for 3 h. 47.34 mg of G5.NH 2 Dissolved in DMSO solution and mixed thoroughly according to the molar ratio of 10:1, and reacted for 72h. Add 41.39 mg HAuCl to the product of the above reaction according to the molar ratio of dendrimer to gold of 1:50 4 React for 30min, then add 18.9mg NaBH 4 The reaction was stirred for 3h. After 3 days of dialysis, {(Au 0 ) 50 -G5.NH 2 -mPEG 10}, recorded as S1 (such as Figure 17 a). Weigh 11.26 mg of FA, 4.4 mg of EDC and 2.64 mg of NHS and mix uniformly according to the ratio of 1:0.9:0.9. Add 20.25 mg of PEG and activated FA to 47.34 mg of G5.NH in a molar ratio of 1:2 2 In, avoid light and stir for 72h. Then add HAuCl 4 After reacting for 30 minutes, quickly add 5 times the amount of NaBH 4 It was reduced and stirred for 3h u...

Embodiment 2

[0054] Weigh 5mg of S1 and S2 respectively and dissolve in 700mL of D 2 In O, the two solutions were sonicated, and then the H NMR spectra of the two complexes were measured using an NMR spectrometer. NMR results such as figure 1 as shown, 1 H NMR spectrum was used to characterize the grafted G5.NH 2 The number of carbon chains on the G5.NH 2 The proton peak of the characteristic group is between 2.0-3.5ppm in chemical shift, and the proton absorption peak appears between 3.4-3.6ppm, indicating that mPEG is successfully connected to G5.NH 2 Above, the proton absorption peaks appearing between 6.6-8.5ppm indicate that FA was successfully connected to G5.NH 2 superior. Use the origin software to calculate the integral area of ​​the proton peak, and get the average of each G5.NH 2 There are about 10 mPEGs and about 3.2 FAs attached to the surface. The results showed that {(Au 0 ) 50 -G5.NH 2 -mPEG 10} and {(Au 0 ) 50 -G5.NH 2 -PEG 10 -FA 5}.

Embodiment 3

[0056] Material S1 and material S2 were respectively dissolved in ultrapure water to prepare a solution with a concentration of 200 μg / mL, and the ultraviolet absorption value in the range of 300nm-800nm ​​was detected by an ultraviolet-visible spectrophotometer. The results of UV-Vis spectrophotometry were as follows: figure 2 As shown, in the complex G5.NH 2 There are absorption peaks around 520nm, indicating that the gold nanoparticles are successfully wrapped. There is an absorption peak at 280nm, indicating that FA has successfully received G5.NH 2 superior.

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Abstract

The invention discloses an application method for gene transfection by using a polyamide-amine dendritic polymer and gold nano-particle compound as a non-viral vector. The surface of the dendritic polymer is the amino terminal, and the chemical modification is executed to the amino terminal so that the toxicity of the dendritic polymer can be reduced and the transfection efficiency is improved; the terminal of the dendritic polymer is connected with a targeted group, comprising a folic acid, RGD, and a hyaluronic acid, and the gene transfer can be efficiently executed. The prepared polyamide-amine dendritic polymer and gold nano-particle compound has the good gene transfection effect. The transfection process is easily operated, the biocompatibility is good, and the immunogenicity is low. The method can provide the reference for the application of the non-viral vector on the gene transfer.

Description

technical field [0001] The invention relates to a method for applying a compound of polyamide-amine dendrimers and nano-gold particles as a non-viral carrier for gene transfection, in particular to a polyamide-amine dendrimer modified with polyethylene glycol and folic acid The invention relates to an application method of carrying out gene transfection and immunological safety research using molecularly encapsulated nano-gold particles as a non-viral carrier, and belongs to the technical field of gene transfection vectors of functional polymer nanomaterials. Background technique [0002] Gene therapy refers to the method of treating or preventing diseases through manipulation at the gene level. At present, gene therapy is mainly aimed at the treatment of diseases that seriously threaten human health, and cancer is its main application field. Gene therapy technology is developing rapidly, and some gene therapy programs have entered the stage of clinical trials. During clin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C08G73/02
CPCC08G73/028C12N15/87
Inventor 曹雪雁史向阳徐琲李爱军郝欣欣
Owner DONGHUA UNIV
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