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Method for preparing polyethylene glycol-modified polyamide-amine dendrimer macromolecule-wrapped gold nanoparticle-loaded CpG ODN

A technology of amine dendrimer and gold nanoparticles, which is applied in the field of preparation of CpGODN loaded by polyamide-amine dendrimer wrapped with gold nanoparticles, and achieves good gene loading effect, easy operation and good biocompatibility.

Inactive Publication Date: 2019-08-06
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Retrieval of relevant literature and patents at home and abroad showed that polyamide-amine dendrimers coated with polyethylene glycol encapsulated gold nanoparticles were used as gene carriers to load CpG ODN, stimulate the maturation of dendritic cells, and then utilize mature dendritic cells Activation of T cells for immunotherapy of cancer has not been reported yet

Method used

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  • Method for preparing polyethylene glycol-modified polyamide-amine dendrimer macromolecule-wrapped gold nanoparticle-loaded CpG ODN
  • Method for preparing polyethylene glycol-modified polyamide-amine dendrimer macromolecule-wrapped gold nanoparticle-loaded CpG ODN
  • Method for preparing polyethylene glycol-modified polyamide-amine dendrimer macromolecule-wrapped gold nanoparticle-loaded CpG ODN

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Experimental program
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Effect test

Embodiment 1

[0062] Weigh 18.55 mg of mPEG-COOH (molecular weight: 2000) and dissolve it in DMSO, mix 17.78 mg of EDC and mPEG-COOH solution uniformly at a ratio of 10:1, and then add 10.67 mg of NHS to react for 3 hours. 24.13 mg of G5.NH 2 Dissolve in DMSO solution and mix well according to the ratio of 20:1, and react for 72 hours. Add 636.78 μl HAuCl to the product of the above reaction according to the molar ratio of dendrimers to gold of 1:25 4 React for 30 minutes, then add 0.88mg NaBH 4 The reaction was carried out for 3 hours with stirring. After 3 days of dialysis, {(Au 0 ) 25 -G5.NH 2 -mPEG 20}, get the material.

Embodiment 2

[0064] Weigh 5mg of the material and dissolve in 700mL of D 2 O, the solution was sonicated, and then the NMR spectrum of the material was measured using a 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-COOH is successfully connected to G5.NH 2 Above, use the origin software to calculate the integral area of ​​the proton peak, and get the average of each G5.NH 2 About 16.9 mPEG-COOH are attached to the surface. The results showed that {(Au 0 ) 25 -G5.NH 2 -mPEG 20}.

Embodiment 3

[0066] The material was 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: image 3 As shown, G5.NH in the material 2 There are absorption peaks around 520nm, indicating that the gold nanoparticles are successfully wrapped.

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Abstract

The invention relates to a method for preparing polyethylene glycol-modified polyamide-amine dendrimer macromolecule-wrapped gold nanoparticle-loaded CpG ODN and a method for activating immune cells,wherein the method comprises the steps: dissolving polyamide-amine dendrimer macromolecules in a solvent, adding mPEG-COOH, then adding a chloroauric acid solution and a sodium borohydride solution successively, dialysing and freeze-drying, and thus obtaining [(Au<0>)25-G5.NH2-mPEG20]; and then compounding with CPG ODN, and thus obtaining the product. The method has the advantages of short preparation period and easy operation; the material has the advantages of low toxicity in a use dosage, good biocompatibility and good compression of CpG ODN and can effectively stimulate maturation of BMDCcells, the mature BMDC cells can effectively activate T cells, and reference is provided for application of non-viral vectors in cancer immunotherapy.

Description

technical field [0001] The invention belongs to the field of tumor immunotherapy, and particularly relates to a preparation method of polyethylene glycol-modified polyamide-amine dendrimers encapsulating nano-gold particles and loading CpG ODN. Background technique [0002] Cancer is one of the major diseases threatening our country's health. Its incidence rate is now the highest cause of death among Chinese residents. Early detection, early diagnosis and early treatment of cancer is one of the main strategies to reduce mortality and improve survival rate. At present, surgical treatment, radiotherapy and chemotherapy are commonly used methods for malignant tumors, but there are problems such as tumor metastasis and adverse reactions of radiotherapy and chemotherapy. Due to the rapid development of immunology and tumor biology research, immunotherapy has become the fourth tumor treatment method after the three classic tumor treatment methods, and was selected as one of the ...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/34A61K47/02A61K47/10A61K31/711A61P35/00C12N5/0783C12N5/0784
CPCA61K9/5115A61K9/5146A61K31/711A61P35/00C12N5/0636C12N5/0639C12N2501/056C12N2502/1121
Inventor 曹雪雁史向阳陈环郝欣欣
Owner DONGHUA UNIV
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