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Method for preparing CT/MR dual-mode imaging contrast agent based on hyperbranched polyethyleneimine molecules

A technology of polyethyleneimine and dual-modal imaging, which is applied in the preparation of X-ray contrast agents, preparations for in vivo tests, pharmaceutical formulations, etc., can solve the problem of preparation and application of CT/MR dual-modal imaging contrast agents. And other issues

Inactive Publication Date: 2015-01-07
DONGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A search of domestic and foreign literature and patents on CT / MR dual-mode imaging contrast agents shows that: at present, no CT / MR dual-mode imaging based on hyperbranched polyethyleneimine molecules wrapped with gold nanoparticles and chelated gadolinium ions has been found. Report on preparation and application of state imaging contrast agent

Method used

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  • Method for preparing CT/MR dual-mode imaging contrast agent based on hyperbranched polyethyleneimine molecules
  • Method for preparing CT/MR dual-mode imaging contrast agent based on hyperbranched polyethyleneimine molecules
  • Method for preparing CT/MR dual-mode imaging contrast agent based on hyperbranched polyethyleneimine molecules

Examples

Experimental program
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Embodiment 1

[0064] (1) In 5 mL of mPEG-COOH in DMSO (12 mg / mL), add 5 mL of EDC in DMSO (11.5 mg / mL), stir for 0.5 h, then add 3 mL of NHS in DMSO (11.5 mg / mL), Continue to stir the reaction for 3h. It was then added dropwise to 10 mL of PEI in DMSO (2.5 mg / mL), and the reaction was stirred for 3 d. 5 mL DOTA-NHS in DMSO (1.52 mg / mL) was added dropwise to the above mixed solution, and the reaction was stirred for 1d. First, it was dialyzed against PBS buffer solution for 1d (3×2L), then dialyzed against deionized water for 2d (6×2L), and freeze-dried to obtain white solid powder PEI-mPEG-DOTA.

[0065] (2) Take 50mg PEI-mPEG-DOTA (0.25mg / mL, ultrapure water as solvent), stir and mix well, add 5.31mL HAuCl dropwise 4 Aqueous solution (10mg / mL) was stirred at room temperature for 30min, the solution turned light yellow, then 5mL NaBH was added 4 Ice solution (4.88mg / mL), the solution turned dark red instantly, and continued to stir at room temperature for 3h;

[0066] (3) Add 1mL Gd(NO ...

Embodiment 2

[0069] The ultraviolet absorption value of the Gd-Au PENPs solution prepared in Example 1 prepared by 0.1 mg / mL of ultrapure water prepared in different temperatures (4, 25, 37 and 50° C.) was measured with an ultraviolet absorption spectrometer, and it was found that the Gd-Au PENPs solution Under different temperature conditions, the SPR peaks of gold nanoparticles are all located at about 520nm, and there is no obvious shift. Similarly, the UV absorption value of the Gd-Au PENPs solution prepared in Example 1 prepared by the buffer solution of 5, 6, 7 and 8 with a pH of 5, 6, 7 and 8 was measured with an ultraviolet absorption spectrometer, and it was found that the Gd-Au PENPs solution was prepared in different The SPR peaks of gold nanoparticles do not shift significantly under pH conditions. Illustrate that the Gd-Au PENPs material prepared by the present invention has good stability under different temperature and pH conditions. Figure 5 .

Embodiment 3

[0071] The cytotoxicity of the Gd-Au PENPs prepared in Example 1 was detected by MTT assay. Weigh 1 mg of Gd-Au PENPs, prepare 1 mg / mL PBS solution, and sterilize overnight with ultraviolet irradiation. Then prepare sterile nanoparticle colloids with Au concentrations of 0, 100, 250, 500, 1000, and 2000 μM with sterile PBS on the ultra-clean workbench (the material is added to the medium, and its concentration is diluted 10 times). KB cells (10,000 cells per well) were planted in 96-well plates overnight to allow them to adhere to the wall, and then culture medium containing different concentrations of Gd-Au PENPs was added for another 24 hours. After 24 hours, add 20 μL MTT to each well, continue to culture at 37°C for 4 hours, then discard the culture medium, add 150 μL DMSO, shake for 15 minutes, measure the UV absorption value at 570 nm with a microplate reader, and calculate based on this value Cell viability (attached Figure 7 ). It was found that the cell viability ...

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Abstract

The invention relates to a method for preparing a CT / MR dual-mode imaging contrast agent based on hyperbranched polyethyleneimine molecules. The method comprises the following steps: (1) adding an m-PEG-COOH solution activated by EDC / NHS into a hyperbranched polyethyleneimine solution, adding a DOTA-NHS solution, stirring, and reacting, thereby obtaining the functionalized polyethyleneimine PEI-mPEG-DOTA; (2) adding an HAuCl4 solution into the aqueous solution of PEI-mPEG-DOTA, reducing by using a NaBH4 solution, adding a Gd(NO3)3 solution, continuously stirring, and finally adding triethylamine and acetic anhydride, and reacting for 20-30 hours; and (3) dialyzing the solution obtained in the step (2), performing freeze drying, thereby obtaining the CT / MR dual-mode imaging contrast agent. According to the method disclosed by the invention, the low-price and readily available polyethyleneimine molecules are taken as carriers, so that the cost of the material is reduced, and the prepared CT / MR dual-mode imaging contrast agent has high biocompatibility and has good in-vivo and in-vitro CT and MR imaging effects.

Description

technical field [0001] The invention belongs to the field of preparation of nano-contrast agents, in particular to a preparation method of CT / MR dual-mode imaging contrast agents based on hyperbranched polyethyleneimine molecules. Background technique [0002] Due to its low price, high spatial resolution, short image acquisition time, and the ability to provide high-resolution 3D slice information, CT imaging technology has become the most common tool for clinical disease diagnosis, but CT imaging There are also some problems that need to be solved urgently, such as poor soft tissue resolution, high radioactive radiation during the detection process, and kidney toxicity when traditional iodine-containing small molecule CT contrast agents are used at high concentrations. MR imaging technology is a molecular imaging method developed rapidly with the development of integrated circuit technology and superconductor technology. It has high soft tissue resolution and sensitivity, ...

Claims

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Application Information

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IPC IPC(8): A61K49/04A61K49/12
Inventor 史向阳周本青汤光宇彭琛付繁繁
Owner DONGHUA UNIV
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