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Method for preparing RGD peptide targeted ultra-small ferriferrous oxide MRI positive nanoprobe

A technology of ultra-small iron ferric oxide and ferric iron tetroxide, applied in preparations for in vivo tests, pharmaceutical formulations, etc., can solve problems such as difficult biological functionalization and no discovery, and achieve increased water solubility and biophase Capacitive, easy to operate, easy to prepare

Inactive Publication Date: 2015-08-12
DONGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It has been reported in the literature that ultra-small iron oxide nanoparticles can be used as T 1 Enhanced MRI contrast agent, but its synthesis method is generally an oil phase pyrolysis method, the disadvantage is that the particles need to be transferred from the oil phase to the water phase, and it is difficult to implement biofunctionalization on the particle surface (Sun et al., J.Am .Chem.Soc.2008,7542–7543)
[0003] Searching the domestic and foreign literature, there is no relevant information about the MRI positive contrast agent prepared by one-step solvothermal method to prepare ultra-small ferric oxide nanoparticles modified with PEG-RGD cyclic polypeptide and the targeted MRI diagnosis of tumor models in vivo. to report

Method used

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  • Method for preparing RGD peptide targeted ultra-small ferriferrous oxide MRI positive nanoprobe
  • Method for preparing RGD peptide targeted ultra-small ferriferrous oxide MRI positive nanoprobe
  • Method for preparing RGD peptide targeted ultra-small ferriferrous oxide MRI positive nanoprobe

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Embodiment 1

[0067] 1.09g of ferric chloride hexahydrate was dissolved in 40mL of diethylene glycol (also known as diethylene glycol, DEG), and then 0.47g of sodium citrate (Na 3 Cit) was dissolved in the above solution, and stirred at 80° C. for 1 hour under an air atmosphere. After the sodium citrate was completely dissolved, 1.312 g of anhydrous sodium acetate was added to the above solution, and the stirring was continued until the sodium acetate powder was completely dissolved, and then Transfer the solution to a 50mL autoclave, and react at 200°C for 4 hours; after the reaction, cool to room temperature naturally, transfer the product to a 50mL centrifuge tube and centrifuge at 8500rpm for 15 minutes, discard the supernatant, and return to the solution with absolute ethanol. Dissolve, centrifuge at 8500rpm for 15 minutes, repeat the operation 3 times, and then dry the precipitate at 60°C to obtain ultra-small ferric oxide nanoparticles; from the surface potential and hydrated particle...

Embodiment 2

[0073] Get the Fe prepared by embodiment 1 respectively 3 o 4 , Fe 3 o 4 -Fe prepared by PEG-RGD nanoparticles and comparative example 1 3 o 4 -Dissolve 2 mg of PEG nanoparticles in ultrapure water to obtain a nanoparticle suspension, which is homogenized by ultrasound, and the surface potential and hydrated particle size are measured. The experimental results show that the prepared Fe 3 o 4 , Fe 3 o 4 -PEG-RGD and Fe 3 o 4 The surface potentials of -PEG nanoparticles were -39.7, -10.1 and -8.8mV; the hydrated particle diameters were 14.6, 212.5 and 168.7nm. From the changes of surface potential and hydrated particle size, it can be concluded that PEG or PEG-RGD has been successfully modified on the surface of Fe3O4 nanoparticles.

[0074] Take by weighing two kinds of materials that embodiment 1 prepares respectively: Fe 3 o 4 , Fe 3 o 4 The control group material Fe obtained by -PEG-RGD and comparative example 1 3 o 4 -PEG 2mg for infrared spectroscopy test ...

Embodiment 3

[0078] In order to ensure that the nanoparticles prepared by the present invention can be safely used for in vivo bioimaging diagnosis, the prepared Fe 3 o 4 -PEG-RGD nanoparticles and control material Fe 3 o 4 -PEG blood compatibility was evaluated. Calculate and weigh Fe according to the iron concentration calculation of two kinds of materials measured in embodiment 2 3 o 4 -PEG-RGD nanoparticles (embodiment 1) and contrast material Fe 3 o 4 -Two kinds of nanoparticles with 1 mg of total iron in PEG (comparative example 1) were respectively dispersed in PBS to prepare a concentration of 1 mg / mL as the mother liquor, and then the concentrations of 10 μg / mL, 20 μg / mL, and 40 μg were sequentially prepared in PBS / mL, 60 μg / mL and 80 μg / mL nanoparticle suspensions. Take an appropriate amount of human fresh blood, first centrifuge (2000rpm, 5 minutes) to remove the supernatant, then wash the red blood cells 5 times with PBS, collect healthy red blood cells and dilute them ...

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Abstract

The invention relates to a method for preparing an RGD peptide targeted ultra-small ferriferrous oxide MRI positive nanoprobe. The method comprises the steps of: conducting a one-step solvothermal synthesis method to obtain surface sodium citrate stabilized ultra-small Fe3O4 nanoparticles, then modifying NH2-PEG-RGD to the nanoparticle surface to obtain the RGD targeted ultra-small ferriferrous oxide MRI positive molecular probe. The PEG-RGD modified ultra-small ferriferrous oxide nanoprobe has long cycle time in mice, and realizes targeting T1-weighted enhancement MRI on the surface of cancer cells with alphavbeta3 integrin high expression (U87MG cells, human brain glioma cells) and subcutaneously transplanted tumor on animal level and cellular level. The Fe3O4 nanoparticles prepared by the invention can be in stably dispersed in an aqueous solution for a long time, and do not agglomerate. The preparation method provided by the invention has the advantages of simpleness, low cost and potential in industrialization and commercialization.

Description

technical field [0001] The invention belongs to the field of preparation of MRI nano-probes, in particular to a preparation method of ultra-small iron ferric oxide MRI-positive nano-probes targeted by RGD polypeptides. Background technique [0002] Cancer, also known as malignant tumor in medical terms, has directly or indirectly affected the lives of many people and has become the number one killer threatening human health. Therefore, early diagnosis and treatment become the key to cure cancer. In terms of early diagnosis of tumors, traditional imaging techniques can only understand tumor volume and anatomical location, while molecular imaging techniques can obtain more detection parameters, such as evaluation of tumor growth kinetics, detection of molecular abnormalities before malignant transformation, tumor cell Markers, etc., and in vivo molecular imaging can realize the study of pathogenesis without damaging the microenvironment of organisms, and help decipher complex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/14A61K49/12
Inventor 史向阳罗宇张贵祥杨嘉
Owner DONGHUA UNIV
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