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A preparation method of zwitterions and morpholine-modified polyamide-amine dendrimers wrapped with gold nanoparticles

A technology of amine dendrimers and nano-gold particles, which is applied in the field of preparation of polyamide-amine dendrimers to achieve simple reaction conditions, simple transfection conditions, and good gene transfection effects

Active Publication Date: 2021-02-26
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Retrieval of related literature and patents at home and abroad shows that the method of using zwitterions and morpholine-modified PAMAM dendrimers wrapped with gold nanoparticles as a carrier for gene transfection has not been reported yet.

Method used

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  • A preparation method of zwitterions and morpholine-modified polyamide-amine dendrimers wrapped with gold nanoparticles
  • A preparation method of zwitterions and morpholine-modified polyamide-amine dendrimers wrapped with gold nanoparticles
  • A preparation method of zwitterions and morpholine-modified polyamide-amine dendrimers wrapped with gold nanoparticles

Examples

Experimental program
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Effect test

Embodiment 1

[0045] (1) With 10mL of anhydrous propanol as a solvent, β-propiolactone (2.97g) and N-[(3-dimethyl)propyl]acrylamide (DMAPA, 4.62g) were dissolved in anhydrous, N 2 And stirring and reacting at 0°C for 3 hours, suction filtration, washing with anhydrous acetone three times, 5 mL each time, and vacuum drying to obtain a white powder of carboxybetaine acrylamide (CBAA).

[0046] (2) Weigh 10 mg of the fifth generation polyamide-amine dendrimer G5.NH 2 and 16.9mg of CBAA were dissolved in 5mL of NaCl solution (0.138M) and 5mL of methanol respectively. After each was completely dissolved, they were stirred together and reacted for 3 days to obtain G5.NH 2 - CBAA solution. Dialyze with a dialysis bag with a molecular weight cut-off of 8000-14000 for three days (dialyze in PBS buffer for one day, change the solution 3 times; dialyze in distilled water for 2 days, change the water 6 times), freeze-dry to obtain dry G5.NH 2 -CBAA.

[0047] (3) Weigh 1.29 mg of (R)-morpholine-3-car...

Embodiment 2

[0051] HOOC-PEG-Morpholine, G5.NH prepared in Example 1 2 -CBAA, Mor+ and the prepared {(Au 0 ) 25 -G5.NH 2 -CBAA-mPEG} (denoted as Mor-) for characterization. 1 H NMR characterization results as figure 1 as shown, figure 1 The proton peak of -NH-CO- appeared at 6.32ppm in (a), indicating that PEG and HCl.Morpholine-COOH were successfully connected through -NH-CO-, and 0.25 Morpholines were connected to each PEG through integration. figure 1 (b) for G5.NH 2 -CBAA's proton NMR spectrum, the characteristic peak of CBAA appears at 1.89ppm, after integration, it is found that each G5 is loaded with 18.5 CBAA molecules on average. figure 1 (c) is {(Au prepared in comparative example 1 0 ) 25 -G5.NH 2 -CBAA-mPEG} H NMR spectrum, the characteristic peak of mPEG appears at 3.5-3.8ppm, after integration, it is found that each G5 is loaded with 15.3 mPEG molecules on average. figure 1 (d) for G5.NH 2-CBAA-PEG-Morpholine's H NMR spectrum, the characteristic peak of –CH2- ...

Embodiment 3

[0053] The two materials Mor+ in Example 1 and Mor- in Comparative Example 1 were used to prepare vector / pDNA complexes with different N / P, and gel retardation experiments were performed. 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: 0.25:1, 0.5:1, 1:1, 2:1, 4:1, 8:1. 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 4 As shown, both materials can well complex with two pDNAs at a lower N / P (N / P=0.5), which blocks the migration of pDNAs. The ability to compress and wrap DNA ...

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Abstract

The invention relates to a preparation method of zwitterions and morpholine-modified polyamide-amine dendrimers that wrap nano-gold particles, comprising: preparation of carboxybetaine acrylamide CBAA, G5.NH 2 Preparation of ‑CBAA, preparation of zwitterion and morpholine-modified polyamide‑amine dendrimers solution, preparation of zwitterions and morpholine modified polyamide‑amine dendrimers wrapped with gold nanoparticles. The surface of the dendrimer prepared by the present invention has abundant amino groups, has good gene transfection effect, and provides an application basis for constructing a safe and efficient gene carrier for gene therapy; the reaction conditions of the present invention are simple, and the synthesis and preparation are easy to operate. The obtained zwitterions and morpholine-modified polyamidoamine dendrimers wrapped with gold nanoparticles are used as carriers for gene transmission, with simple transfection conditions and high transfection efficiency, and have potential applications in gene therapy of cancer, etc. prospect.

Description

technical field [0001] The invention belongs to the field of preparation methods of functional polymer nanomaterial carriers, in particular to a preparation method of zwitterions and morpholine-modified polyamide-amine dendrimers wrapping nano-gold particles. 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 key to gene therapy is to find safe and efficient gene carriers. Usually, the carriers used for gene delivery include viral vectors and non-viral vectors. Although the research on ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G73/02C08K3/08C12N15/87A61P35/00
CPCA61P35/00C08G73/028C08K3/08C08K2003/0831C08K2201/011C12N15/87
Inventor 史向阳熊智娟沈明武
Owner DONGHUA UNIV
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