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Preparation method of RGD-modified subminiature superparamagnetic iron oxide nanoparticles

A technology of superparamagnetic iron oxide and nanoparticles, which can be used in preparations for in vivo tests, pharmaceutical formulations, emulsion delivery, etc., can solve problems such as no discovery, and achieve simple and easy operation, low raw material cost, high target tropism effect

Inactive Publication Date: 2015-03-11
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Searching the literature at home and abroad, there is no relevant report on the preparation of USPIO nanoprobes with ultra-high relaxation rate RGD-specific targeting ability by reduction method for targeted MR imaging diagnosis of brain glioma models in vitro and in vivo

Method used

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  • Preparation method of RGD-modified subminiature superparamagnetic iron oxide nanoparticles
  • Preparation method of RGD-modified subminiature superparamagnetic iron oxide nanoparticles
  • Preparation method of RGD-modified subminiature superparamagnetic iron oxide nanoparticles

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Effect test

Embodiment 1

[0063] 1.30g FeCl 3 ·6H 2 O was dissolved in 20mL of ultrapure water, transferred to a 250mL three-necked flask and stirred; after blowing nitrogen for 15 minutes, dissolved 0.2g of sodium sulfite in 10mL of ultrapure water and added to the three-necked flask, and continued stirring for 30 minutes; then 0.5g of Aqueous solution of hyperbranched polyethyleneimine PEI and 2 mL of NH 3 ·H 2 O was also added to the three-necked flask, and stirred at 70°C for 30 minutes; then reacted at room temperature for 1.5 hours; after the reaction, the obtained black USPIO / PEI.NH 2 Magnetic separation, remove the supernatant, add an appropriate amount of ultra-pure water for ultrasonic dispersion, and then magnetic separation, repeat the ultra-pure water washing three times to remove impurities, and then re-disperse in 20mL ultra-pure water to obtain PEI-coated USPIO Nanoparticles (USPIO / PEI.NH 2 ). At the same time, bare USPIO was prepared as a control in the absence of PEI by the same ...

Embodiment 2

[0065] Take 20mL USPIO / PEI.NH 2 (154 mg) (Example 1) was washed with DMSO and redispersed in 20 mL of DMSO solution. Next, 2 mL of FI solution (3.6 mg) dissolved in DMSO was added to USPIO / PEI.NH 2 DMSO solution, stirred for 1 day, the product USPIO / PEI.NH 2 -FI is magnetically separated and washed with ultrapure water, then dispersed into water. Take 25μL USPIO / PEI.NH respectively 2 (Example 1), USPIO / PEI.NH 2 Put the aqueous solution of -FI in a 2mL centrifuge tube, then add 700μL ultrapure water to it, ultrasonically homogenize, and measure the ultraviolet absorption (see appendix figure 2 ). As can be seen from the figure, USPIO / PEI.NH 2 There is no obvious ultraviolet absorption peak at 400 to 600nm, while USPIO / PEI.NH 2 -FI has an obvious UV absorption peak at 500nm, which shows that FI has been successfully modified to USPIO / PEI.NH 2 nanoparticle surface.

Embodiment 3

[0067] 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 vacuum 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 attached image 3 ). 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 0.5 RGD is connected to each PEG.

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Abstract

The invention relates to a preparation method of RGD-modified subminiature superparamagnetic iron oxide nanoparticles. The preparation method comprises the following steps of preparing PEI-coated subminiature superparamagnetic iron oxide nanoparticles by a mild reduction method, modifying surfaces of USPIO / PEI.NH2 nanoparticles by fluorescein isothiocyanate (FI) and a RGD compound (COOH-PEG-RGD) subjected to pegylation, and carrying out acetylation treatment on the surfaces of the nanoparticles. The preparation method has simple processes and mild reaction conditions and can be operated easily. The RGD-modified subminiature superparamagnetic iron oxide nanoparticles have small sizes, a high relaxation rate, good colloid stability and biocompatibility, and high alpha<v>beta<3> integrin high-expression tumor cell targeting, and have a latent application value in the field of in-vivo tumor targeting MR imaging diagnosis.

Description

technical field [0001] The invention belongs to the field of preparation of nuclear magnetic resonance imaging contrast agents, in particular to a method for preparing RGD-modified ultra-small superparamagnetic iron oxide nanoparticles. Background technique [0002] In recent years, superparamagnetic Fe3O4 nanoparticles have been widely used in the field of biomedicine, especially as contrast agents for magnetic resonance imaging (MRI). Our research group has prepared two kinds of ferric oxide nanoparticles with different surface modifications through a simple hydrothermal synthesis method, and explored the feasibility of their application in MR imaging diagnosis (Shen Mingwu, Cai Hongdong, Shi Xiangyang. An APTS Preparation method of modified iron oxide magnetic nanoparticles. Patent No.: ZL 2011 1 0104443.1; Cai et al., ACS Appl.Mater.Interfaces 2013, 5(5), 1722–1731; 37. Shi Xiangyang, Cai Hongdong, Shen Mingwu. A preparation method of HPEI-wrapped iron oxide magnetic na...

Claims

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Application Information

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IPC IPC(8): A61K49/18A61K49/14
Inventor 沈明武胡勇李静超韦平史向阳
Owner DONGHUA UNIV
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