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Preparation method and application of ferroferric oxide brain-targeted contrast agent

A contrast agent and reaction technology, which can be used in MRI/MRI contrast agents, preparations for in vivo experiments, pharmaceutical formulations, etc. The effect of enhancing the T2 dark effect, the preparation method is simple and easy to operate, and the application prospect is broad

Pending Publication Date: 2021-09-24
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most contrast agents are administered through intravenous injection, and it is difficult to pass through the blood-brain barrier, so there are still some defects in the diagnosis of brain diseases

Method used

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  • Preparation method and application of ferroferric oxide brain-targeted contrast agent
  • Preparation method and application of ferroferric oxide brain-targeted contrast agent
  • Preparation method and application of ferroferric oxide brain-targeted contrast agent

Examples

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Embodiment 1

[0031] Example 1 Expression and purification of cholera toxin subunit B

[0032]Take out the glycerol tube strain CTB stored in the -20°C refrigerator, inoculate it on a clean bench, and culture it overnight at 37°C on a shaker at 220 rpm for 12 hours. After the overnight culture, scale-up culture in Erlenmeyer flasks was carried out, and cultured on a shaker at 37° C. and 220 rpm for 4 hours. Next, IPTG was added to induce protein expression, and cultured on a shaker at 37° C. and 220 rpm for 5 h. Finally, collect the bacteria by centrifugation, discard the supernatant and save the precipitate. Add lysis buffer to the collected wet bacterial pellet, and stir overnight at 4°C until the bacterial cells in the solution are completely suspended. Freeze and thaw repeatedly between -80°C and 37°C to completely lyse the bacteria, then centrifuge, discard the supernatant and save the precipitate. Wash the precipitate with washing solution repeatedly to remove impurities. After was...

Embodiment 2

[0034] Example 2 Fe 3 o 4 @SiO 2 preparation and characterization

[0035] Take a 50mL plastic centrifuge tube and weigh 10mg of dry Fe 3 o 4 Add 2 mL of 0.1 mol / L dilute hydrochloric acid to the tube, and then use 100% power to ultrasonically vibrate for 10 min to make it fully mixed. After the sonication, the precipitate was adsorbed with a magnet, the supernatant was poured out and discarded, and the precipitate was washed three times with deionized water. Add 20mL of absolute ethanol, 5mL of deionized water, and 0.25mL of NH3·H2O to the sediment in the centrifuge tube in sequence, and use 100% power to ultrasonically vibrate for 10 minutes to make it fully mixed. After the sonication, 120 μL of tetraethyl silicate (TEOS) was added, placed in a shaker at 220 rpm, and shaken at room temperature for 12 hours. After the reaction, the product was adsorbed by a magnet, the supernatant was poured off, washed three times with absolute ethanol, and dried in vacuum. After dry...

Embodiment 3

[0043] Example 3 Fe 3 o 4 @SiO 2 Coupling with CTB to prepare T 2 contrast agent

[0044] Accurately weigh 50 mg dry Fe 3 o 4 @SiO 2 The magnetic nanoparticles were placed in a 50mL round-bottomed centrifuge tube, and then 20mL of PBS buffer (pH=6) was added, sonicated, and dispersed for 30min. After the sonication, 2 mL of EDC·HCl (2 mg / mL, 0.021 mmol) was added thereto, followed by sonication, and reacted for 20 min. After the end, 1.2ml of NHS (1mg / mL, 0.010mmol) was added, and then further ultrasonic treatment was performed, and the reaction was carried out for 20min. After the reaction of EDC and NHS on the magnetic nanoparticles, 18.9 mL of CTB buffer solution (0.3 mg / mL in PBS, pH=6) was added to the above reaction mixture and shaken at 4°C and 22 rpm for 6 h. After the reaction, the supernatant was separated by magnetic adsorption, placed in another clean centrifuge tube for storage, washed carefully with PBS buffer, and dried in vacuum after washing. Fourier ...

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Abstract

The invention belongs to the technical field of inorganic material binding protein, and particularly relates to preparation and application of a T2 contrast agent based on Fe3O4. The preparation method comprises the following steps of modifying Fe3O4 on the surface of SiO2, coupling the modified Fe3O4 with CTB through coupling agents NHS and EDC.HCl, and combining the Fe3O4 with the CTB to prepare the T2 contrast agent with relatively high biocompatibility, so that the problem that the contrast agent is difficult to penetrate through a blood brain barrier can be solved, and the brain lesion diagnosis can be better realized.

Description

technical field [0001] The invention belongs to the technical field of inorganic materials binding proteins, in particular to a Fe-based 3 o 4 the T 2 Preparation of contrast agents and their applications. Background technique [0002] Magnetic resonance imaging (MRI) is a non-invasive, non-invasive medical imaging diagnostic technology, which generates NMR signals through hydrogen atoms in fat and water in organisms to provide high-spatial resolution and high-contrast images of soft tissue lesions. . In order to increase the contrast with the background area and improve the signal-to-noise ratio and sensitivity, the participation of MRI contrast agents is often required. MRI contrast agents are mainly divided into T 1 contrast agent and T 2 contrast agent. T 1 Contrast agents can shorten the longitudinal relaxation time, appear bright signals in the target area, and produce bright contrast effects. Currently, gadolinium-based contrast agents based on gadolinium compl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/14A61K49/08
CPCA61K49/14A61K49/08
Inventor 王华倩杨静茹纪美琪何贤樱陈宇
Owner GUANGDONG UNIV OF TECH
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