Preparation method of functionalized gold nanostar/siRNA complex stabilized by RGD-modified dendrimers

A dendrimer, gold nanotechnology, applied in recombinant DNA technology, medical preparations containing active ingredients, and other methods of inserting foreign genetic materials, etc. The effect of monodispersity, simple operation and low cost of raw materials

Inactive Publication Date: 2017-12-26
DONGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Searching the literature at home and abroad, there is no report on the preparation of functionalized gold nanostars stabilized by RGD targeting dendrimers and their use in CT imaging and combined photothermal and gene therapy

Method used

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  • Preparation method of functionalized gold nanostar/siRNA complex stabilized by RGD-modified dendrimers
  • Preparation method of functionalized gold nanostar/siRNA complex stabilized by RGD-modified dendrimers
  • Preparation method of functionalized gold nanostar/siRNA complex stabilized by RGD-modified dendrimers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] (1) Weigh 10mg G3.NH 2 Dissolve in 20mL of ultrapure water, and add 3.25μL of methylthioglycolate (Methylthioglycolate, 95%, Mw=106.14, ρ=1.187) to it, react in a water bath at 60°C for 2 days, and use a dialysis bag with a molecular weight cut-off of 1000 in water Partially mercaptolated G3.NH was obtained after three days of dialysis 2 (G3.NH 2 -SH), and stored at -20°C after lyophilization. Take 3 mg of freeze-dried G3.NH 2 and G3.NH 2 -SH dissolved in deuterated water for NMR test, from figure 1 As can be seen in G3.NH 2 -SH(b) compared to G3.NH 2 (a), there is an additional enhanced absorption peak at 3.25, which is the characteristic peak of the amide bond generated by the reaction between the amino group on the dendrimer and methyl thioglycolate, and it can be known by peak area integration that each G3. NH 2 15.51 sulfhydryl groups are connected on it.

[0056] (2) Weigh 100mg trisodium citrate, dissolve it in 9.9mL ultrapure water, take 1.5mL and add ...

Embodiment 2

[0059] Take the aqueous solutions of Au seeds and Au nanostars prepared in Example 1 respectively in 2mL centrifuge tubes, add 700 μL of ultrapure water therein, ultrasonically uniform, and measure the ultraviolet absorption (see figure 2 , UV absorption spectra of gold seeds and AuNSs at 400 to 1000 nm). The ultraviolet results show that the absorption peak of the gold seed (a) is around 520nm, while the ultraviolet absorption peak of the gold nanostar (b) synthesized by the seed growth method is around 800nm, indicating that the gold nanostar (b) with a unique absorption peak in the near-infrared region has been successfully synthesized. Gold nanostars.

[0060] Take 5 μL of the aqueous solutions of Au seeds and Au nanostars prepared in Example 1, respectively, and then prepare 100 μL of nanoparticle suspension with ultrapure water. Drop 5 μL of the nanoparticle suspension on the surface of the copper grid and dry it in the air for TEM testing (see image 3 , morphology a...

Embodiment 3

[0063] Mix 3.70mg 6-M with 20mg NH 2 -PEG-COOH was mixed in 5mL DMSO solution, stirred and reacted for 10 hours to obtain COOH-PEG-6-M; then 6.91mg RGD fully dissolved in 2mL DMSO was added dropwise to COOH-PEG-6-M In DMSO solution, stirred and reacted for 1 day, dialyzed with a dialysis bag with a molecular weight cut-off of 1000 for 3 days, and then freeze-dried to obtain COOH-PEG-RGD. The COOH-PEG-RGD that gets 3mg synthesis is dissolved in deuterated DMSO and carries out nuclear magnetic resonance analysis ( 1 H NMR) (see Figure 5a). It can be seen from the figure that the spectral peaks at 7.3 and 7.4ppm prove that RGD is successfully connected to PEG, and it can be seen from the peak area integration that 0.35 RGD is connected to each PEG.

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Abstract

The invention relates to a preparation method for a functionalized gold nanostar / siRNA compound modified through RGD and stabilized through dendritic macromolecules. The method comprises the steps that gold nanoparticles are prepared through a sodium citrate reduction method, and AuNSs is prepared through a seed growing method; excessive MTG is dropwise added into a G3.NH2 solution, the excessive MTG and the G3.NH2 solution are subjected to thermostatic reaction to obtain G3.NH2-SH, then the G3.NH2-SH is dropwise added into an AuNSs aqueous solution, and the G3.NH2-SH and the AuNSs aqueous solution are stirred to obtain G3-AuNSs; 6-M and COOH-PEG-NH2 are subjected to reaction to obtain COOH-PEG-MAL; and then SH-RGD and the COOH-PEG-MAL are subjected to reaction to obtain COOH-PEG-RGD, the COOH-PEG-RGD is added into DMSO dispersion liquid of the G3-AuNSs after being activated, and Au DSNSs is obtained through reaction and hatched with VEGF siRNA, so that the functionalized gold nanostar / siRNA compound is obtained. According to the preparation method, the reaction condition is mild, the process is simple, and the method is easy to operate and has potential application value in the diagnosis and treatment integration field.

Description

technical field [0001] The invention belongs to the field of preparation of integrated nanomaterials for diagnosis and treatment, in particular to a method for preparing a functionalized gold nanostar / siRNA complex stabilized by RGD modified dendrimers. Background technique [0002] In recent years, nanomaterials have received increasing attention due to their potential applications in gene / drug delivery, molecular imaging, and targeted therapy. In particular, gold nanoparticles with special morphology are widely used in biosensors, photothermal therapy, and molecular imaging because of their good chemical stability, biocompatibility, catalytic activity, and unique optical properties. In view of the fact that the surface of gold nanomaterials is easily modified by chemotherapeutic drugs and other functional groups, it is often used as a star nano-platform for multimodal therapy and the integration of diagnosis and treatment. [0003] Gold nanostars, as a kind of gold nanopa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B22F9/24C12N15/87A61K49/00A61K41/00A61K48/00A61P35/00B82Y40/00
CPCA61K41/0052A61K48/0008A61K49/0065B22F9/24B82Y40/00C12N15/87
Inventor 沈明武韦平胡勇史向阳
Owner DONGHUA UNIV
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