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Prussian blue nano MRI tracer agent as well as preparation method and application thereof

A Prussian blue and tracer technology, applied in the field of Prussian blue nano-MRI tracer and its preparation, achieves the effects of long-distance transportation, good biocompatibility, biosafety, and good stability

Active Publication Date: 2018-12-21
王金环
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the use of magnetic Prussian blue nanoparticles for targeted diagnosis of Alzheimer's disease has not been reported

Method used

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  • Prussian blue nano MRI tracer agent as well as preparation method and application thereof
  • Prussian blue nano MRI tracer agent as well as preparation method and application thereof
  • Prussian blue nano MRI tracer agent as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1) Preparation of cysteine ​​modified β-amyloid 42 (cys-Aβ42)

[0036] A cysteine ​​containing a sulfhydryl group is inserted between the 28th lysine and the 29th glycine of β-amyloid 42 to synthesize cysteine-modified β-amyloid 42, whose sequence is: DAEFRHDSGYEVHHQKLVFFAEDVGSNK(Cys)GAIIGLMVGGVVIA.

[0037] 2) Preparation of maleimidized Prussian blue nanoparticles (PB@PEI@MAL NPs)

[0038] Dissolve 0.5mM citric acid in 1mM FeCl3·6H2O deionized water solution to prepare mixture A, and dissolve 0.5mM citric acid in 1mM K4Fe(CN)6·3H2O deionized water solution to prepare mixture B, and then heat it at 60℃ Add 20ml of mixed solution B dropwise to 20ml of mixed solution A, and continue to stir for 5 minutes, the mixed solution will turn bright dark blue. Add a polyetherimide (PEI) solution with a concentration of 0.005mM, keep stirring at 60°C for 4 hours, centrifuge at 15000rpm for 30min to collect the precipitate and wash with acetone and deionized water, and finally resuspend...

Embodiment 2

[0044] The Prussian blue nano-NMR tracer PB@PEI@MALcys-Aβ42 NPs prepared in Example 1 was detected and characterized.

[0045] figure 2 It is a transmission electron micrograph of the Prussian blue nano-NMR tracer PB@PEI@MAL-cys-Aβ42 NPs prepared in Example 1.

[0046] The morphology of PB@PEI@MAL-cys-Aβ42 NPs was characterized by transmission electron microscope, the specific operation steps are as follows:

[0047] Dissolve PB@PEI@MAL-cys-Aβ42 NPs in PBS and then disperse ultrasonically. Pipette 10μl of the solution onto a copper mesh covered with ultra-thin carbon film, and fully adsorb for 10-30 minutes until the sample is dry and then proceed in a transmission electron microscope. Observe and take pictures. The result is figure 2 As shown, it can be seen that the particle size of PB@PEI@MAL-cys-Aβ42 NPs is about 60 nm, with a cubic structure, with a relatively uniform particle size distribution and good dispersion.

[0048] image 3 It is the particle size diagram of the Pru...

Embodiment 3

[0056] Figure 5 It is a graph of the relaxation time T1 and T2 with concentration in the MRI of the Prussian blue nano-NMR tracer PB@PEI@MAL-cys-Aβ42 NPs prepared in Example 1.

[0057] Configure PB@PEI@MAL-cys-Aβ42 NPs solution with a concentration gradient of 0-16mg / ml with PBS and place it in a NMR sample tube to evenly disperse; measure the spin of each concentration gradient solution by a 1.5 T NMR analyzer- Lattice relaxation time (T1) and spin-spin relaxation time (T2), while taking grayscale images. Figure 5 The relaxation time T1 and T2 of the weighted NMR imaging of the Prussian blue nano-NMR tracer PB@PEI@MAL-cys-Aβ42 NPs increased with the increase of the concentration of PB@PEI@MAL-cys-Aβ42NPs .

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Abstract

The invention discloses a Prussian blue nano MRI tracer agent as well as preparation method and application thereof, and provides nontoxic Prussian blue nanoparticles-based MRI tracer agent with highspecificity and sensitivity. The maleinimidized Prussian blue nanoparticles combined with cysteine-modified beta-amyloid protein can detect the Abeta deposited in the brain of an Alzheimers patient and clearly image the Abeta on MRI. In the invention, an MRI tracer agent capable of passing through the blood brain barrier and being specifically combined with the Abeta is prepared by adopting Abetahomologous polypeptide as a target of beta amyloid protein in brain and the Prussian blue as an imaging agent. As a detection agent of molecular images, the MRI tracer agent disclosed by the inventioncan provide a new means of AD early diagnosis and curative effect evaluation in the future.

Description

Technical field [0001] The invention belongs to the technical field of biological materials, and specifically relates to a Prussian blue nano-MRI tracer and a preparation method and application thereof. Background technique [0002] Alzheimer's disease (AD) is clinically dominated by intellectual impairment, manifested as senile dementia in middle-aged and elderly people, which seriously threatens the health of middle-aged and elderly people in my country. At present, there is still no effective treatment for AD. With the deepening of research, more and more evidences show that the accumulation of extracellular amyloid (Aβ), the tangles of intracellular nerve fibers and the loss of neurons are significant for AD Pathophysiological characteristics. The main component of amyloid is the enzymatic hydrolysis product of amyloidβ-protein precursor (APP), which is secreted from the cell to the outside of the cell. Based on the "amyloid hypothesis" of the onset of AD, a large number of...

Claims

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Application Information

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IPC IPC(8): A61K49/18A61K49/14B82Y5/00B82Y15/00
CPCA61K49/14A61K49/1866B82Y5/00B82Y15/00
Inventor 王金环常津徐立霞罗仲秋刘雅娟李家林王琼
Owner 王金环
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