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Targeted nano magnetic resonance contrast agent for brain epileptic focus and its preparation and application

A magnetic resonance contrast agent and epilepsy technology, which is applied in the field of molecular imaging to achieve the effect of evaluating prognosis, improving accuracy and safety, and evaluating curative effect

Active Publication Date: 2018-11-16
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the existing diagnostic techniques cannot solve the problem of early and accurate display of epileptic focus from the molecular mechanism level, and the existing contrast agents cannot quantitatively reflect the expression of P-glycoprotein, which is helpful for early detection of drug-resistant epilepsy , in order to help the clinic to improve the accuracy and safety of treatment, and there is no drug delivery system to target the relevant therapeutic drugs to the lesion, so as to realize the integration of imaging and treatment

Method used

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  • Targeted nano magnetic resonance contrast agent for brain epileptic focus and its preparation and application
  • Targeted nano magnetic resonance contrast agent for brain epileptic focus and its preparation and application
  • Targeted nano magnetic resonance contrast agent for brain epileptic focus and its preparation and application

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Embodiment 1

[0048] This embodiment provides a targeted nano-magnetic resonance contrast agent for cerebral epileptic focus and its preparation, as follows:

[0049] Preparation of PEG-USPIO nanoparticles: Accurately weigh 10 mg lipid USPIO, 50 mg DSPE-PEG-MAL, 5 mg DSPE-PEG-FITC, dissolve in 2 ml of dichloromethane, sonicate for 10 min in an ice-water bath, and spin steam at 37°C for 15 min to remove Chloromethane, centrifuged at 2500rpm at 4°C for 10min, discarded the supernatant, dispersed and precipitated with 0.5mL deionized water to obtain nanoparticles.

[0050] Preparation of pepstatin A-PEG-USPIO nanoparticles: add sulfhydryl activating reagent pepstatin A 2mg / ml0.5ml to the prepared deionized water-dispersed nanoparticles 2ml (5mg / mL USPIO) and stir for 4h at low speed , centrifuged at 2500rpm at 4°C for 10min, discarded the supernatant, dispersed and precipitated with 0.5mL 0.01M PBS (pH7.4) to obtain the nanoparticles with targeting functional molecules we needed.

[0051] Use...

Embodiment 2

[0053] BCEC cells (rat brain capillary endothelial cells) were 5×10 5 The concentration of cells in each well was seeded in a six-well plate for experiments; 1-3 wells were the blank control group; 4-6 wells were the experimental group, and 100uM (1.5g / 100ml) glutamic acid was added to each well, and incubated at 37°C for 30min Afterwards, PBS (phosphate buffered saline) was washed twice, and complete culture medium was added for routine culture; 24 hours later, each group was added with PBS, PEG-USPIO, pepstatin A-PEG-USPIO (USPIO concentration was 50 μg / ml), and After 30 minutes, samples were taken for immunofluorescence confocal qualitative analysis and flow cytometry FITC fluorescence quantitative analysis. The results were as follows: Figure 4 , Figure 5 Shown: Figure 4 The indicated glutamate can induce high P-glycoprotein in BCEC cells, Figure 5 The indicated targeting function molecule pepstatin A can occupy the binding site of P-glycoprotein, which is helpful f...

Embodiment 3

[0055] Stereotaxic SD rat left hippocampus CA3 area, microneedle injection of 3 μL kainic acid (KA, 0.5 mg / mL) to establish animal models of epilepsy; injection of 3 μL PBS (phosphate buffer) as the sham operation control group; more than 4% The heart was perfused with polyformaldehyde, and the brain was removed. The expression of P-glycoprotein in the hippocampus of the epilepsy group and the sham operation group was observed by immunofluorescence confocal imaging and Western Blot analysis. The results are as follows: Image 6 Shown: Image 6 It showed that the expression of P-glycoprotein in the hippocampus of the epilepsy group was significantly higher than that of the sham operation group, suggesting that epileptic seizures were closely related to the increased expression of P-glycoprotein.

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Abstract

The invention discloses a targeted nano-magnetic resonance contrast agent for brain epileptic foci, preparation and application thereof. The magnetic resonance contrast agent comprises a target functional molecule, a fluorescent marker and a nano-carrier. The target functional molecule is short peptide that has high affinity to epileptic focus area high expression P-glycoprotein. The fluorescent marker is encapsulated in the nano-carrier by means of covalent connection, and the short peptide is in connection with polyethylene glycol on a nanoparticle surface through covalent connection. The contrast agent can combine the short peptide with the epileptic focus area high expression P-glycoprotein through intravenous injection, promotes gathering of the nano-carrier at epileptic foci, and makes MRI T2 weighted imaged focus area show significant low signal, thereby accurately and visually display idiopathic / cryptogenic epilepsy foci. The contrast agent is conducive to clinical improvement of the treatment accuracy and safety, evaluation of the efficacy of anti-epilepsy treatment means, and assessment of the prognosis of epilepsy patients.

Description

technical field [0001] The invention belongs to the technical field of molecular imaging, and relates to a targeted nano magnetic resonance contrast agent for cerebral epileptic focus and its preparation and application. Background technique [0002] Epilepsy is one of the most common neurological disorders characterized by chronic, spontaneous, and recurrent clinical symptoms. According to statistics, there are about 9 million epilepsy patients in my country, and 450,000 new cases are added every year. About 1 / 3 of them have been treated with regular antiepileptic drugs, but their seizures cannot be controlled, which significantly increases the severity of the disease, mortality and Social and family economic burden. At present, the effect of epileptic foci resection is relatively certain, but clinical practice shows that it is difficult to completely remove the epilepsy, and the morbidity rate is high, which seriously affects the quality of life of patients. One of the im...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/12A61K49/14A61K49/18A61K49/00A61K47/04A61K47/42A61K47/64A61K9/51A61P25/08A61K47/60A61K47/62A61K47/69
Inventor 张军王剑虹于向荣耿道颖庞志清潘嘉炜
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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