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Targeting gene transferring method of folic acid-functionalized PAMAM (polyamidoamine dendrimers) wrapped by gold nanoparticles

A technology for converting polyamidoamine trees and gold nanoparticles, which is applied in the field of targeted gene transfection of folic acid-functional polyamidoamine dendrimer carriers

Inactive Publication Date: 2013-03-13
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Retrieval of related literature and patents at home and abroad shows that the method of targeting gene transfection using folic acid-modified polyamidoamine dendrimers coated with gold nanoparticles has not yet been reported.

Method used

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  • Targeting gene transferring method of folic acid-functionalized PAMAM (polyamidoamine dendrimers) wrapped by gold nanoparticles
  • Targeting gene transferring method of folic acid-functionalized PAMAM (polyamidoamine dendrimers) wrapped by gold nanoparticles
  • Targeting gene transferring method of folic acid-functionalized PAMAM (polyamidoamine dendrimers) wrapped by gold nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Weigh 4.22 mg of folic acid (FA) and dissolve it in 3 mL of DMSO. Weigh 18.33mg EDC and dissolve it in 2ml DMSO, add the EDC solution to the FA solution, and stir for 3h. Weigh the fifth generation polyamidoamine dendrimer (G5.NH 2 ) 40mg, dissolved in 4mL DMSO to prepare a solution with a concentration of 10.0mg / mL. According to FA / G5.NH 2 The molar ratio is 5 / 1, add the activated folic acid solution in (1) to G5.NH 2 Solution, magnetic stirring at room temperature, reaction 3d. Add 0.42mL HAuCl to the above solution 4 The aqueous solution (20mg / mL) was mixed and stirred for half an hour, and then 0.453mL of 10mg / mL NaBH was added 4 Solution (H 2 O:CH 3 OH (volume ratio)=2:1), react for 2h. After the reaction, the reaction product was transferred to a dialysis bag with a molecular weight cut-off of 14,000, and dialyzed in distilled water for two days (2 L×6, three times a day). Then freeze-drying was carried out to obtain the Au DENPs-FA reaction product. The...

Embodiment 2

[0055] The Au DENPs-FA prepared according to the method of Example 1 and the Au DENPs prepared in Comparative Example 1 formed complexes with pDNA, and a gel retardation experiment was performed. Prepare 8 wells of agarose gel (1.0% w / v) containing ethidium bromide (0.1 μg / mL), and place at room temperature until the agarose gel solidifies. Taking 1 μg of pDNA as an example, prepare vector / pDNA complexes according to different N / P (0.125, 0.25, 0.5, 1, 2, 5), and use naked pDNA as a control. Then the corresponding vector / pDNA complexes were respectively added to the wells of the agarose gel with a voltage of 80V and a time of 30min. The migration of DNA in the gel was analyzed using a gel imager. The result is as shown in Figure 3. The results showed that both Au DENPs-FA and Au DENPs could well complex with DNA at a low N / P (N / P=0.5) and block DNA.

Embodiment 3

[0057] After the Au DENPs-FA prepared according to the method of Example 1 and the Au DENPs prepared in Comparative Example 1 were complexed with 5 μg DNA (N / P=1, 2.5, 5), the volume was adjusted to 1 mL with deionized water respectively. The particle size and surface potential were characterized by a Malvern laser particle size analyzer (Malvern, MK, 633nm laser), and the results are shown in Figure 4. The results showed that with the increase of N / P, the sizes of all complexes first decreased and then slightly increased. Under the same N / P, the size of Au DENPs-FA / pDNA complex was not much different from that of Au DENPs / pDNA complex. In addition, with the increase of N / P, the surface potential of Au DENPs-FA / pDNA complexes and Au DENPs / pDNA complexes both increased.

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Abstract

The invention relates to a targeting gene transferring method of folic acid-functionalized PAMAM (polyamidoamine dendrimers) wrapped by gold nanoparticles. The targeting gene transferring method comprises the steps of preparing the folic acid-functionalized PAMAM wrapped by gold nanoparticles; preparing a carrier of the folic acid-functionalized PAMAM wrapped by gold nanoparticles and a plasmid compound; and transferring and expressing targeting cervical cancer cells of the compound. The carrier prepared by the invention is used for transferring genes, has the advantages of moderate transferring condition, easiness in operation, high transferring efficiency, good specificity and the like, and has good application prospects in the aspect of cancer treatment and the like.

Description

technical field [0001] The invention belongs to the field of targeted gene transfection of macromolecular nanocarriers, and in particular relates to a targeted gene transfection method of a folic acid-functionalized polyamidoamine dendritic macromolecule carrier wrapped with nano gold particles. Background technique [0002] At present, as an important means to deal with various genetic diseases and cancers, gene therapy is receiving widespread attention. Generally, gene therapy vectors can be divided into viral vectors and non-viral vectors. Viral vectors are commonly used because of their high transfection efficiency, but they have disadvantages such as immunogenicity, high toxicity, and incapability of large-scale production. Non-viral vectors, especially polyamidoamine dendrimers (PAMAMs), have come from behind and attracted more and more attention due to their relatively low toxicity, high efficiency, and large-scale production. In the treatment of diseases at specifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/34A61K47/22A61K47/02C12N15/87A61K47/54A61K47/59A61K47/69
Inventor 史向阳肖童雨温诗辉
Owner DONGHUA UNIV
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