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Targeted nano-magnetic resonance contrast agent for brain epileptic foci, preparation and application thereof

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

Active Publication Date: 2016-04-20
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 foci, preparation and application thereof
  • Targeted nano-magnetic resonance contrast agent for brain epileptic foci, preparation and application thereof
  • Targeted nano-magnetic resonance contrast agent for brain epileptic foci, preparation and application thereof

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Embodiment 1

[0048] This embodiment provides a targeted nano-magnetic resonance contrast agent for brain epileptic foci and preparation thereof, which are specifically as follows:

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

[0050] Preparation of pepstatin A-PEG-USPIO nanoparticles: add 2ml (5mg / mLUSPIO) of prepared deionized water-dispersed nanoparticles to the sulfhydryl activating reagent pepstatin A2mg / ml0.5ml, then stir at low speed for 4h. After centrifugation at 2500 rpm 4°C for 10 min, the supernatant was discarded, and 0.5 mL 0.01 MPBS (pH 7.4) was dispersed and precipitated to obtain the nanoparticles wit...

Embodiment 2

[0053] Divide BCEC cells (rat brain capillary endothelial cells) into 5×10 5 The concentration of cells in each well is inoculated in a six-well plate for experiment; wells 1~3 are blank control group; wells 4~6 are experimental group, add 100uM (1.5g / 100ml) glutamic acid to each well and incubate at 37℃ for 30min After that, PBS (phosphate buffered saline) was washed twice, and complete culture medium was added for routine culture; after 24 hours, each group was added with PBS, PEG-USPIO, and pepstatin A-PEG-USPIO (USPIO concentration 50μg / ml). After 30 minutes, samples were taken for immunofluorescence confocal qualitative analysis and flow cytometry FITC fluorescence quantitative analysis. The results are as follows Figure 4 , Figure 5 Shown: Figure 4 Shown that glutamate can induce high P-glycoprotein in BCEC cells, Figure 5 The shown targeting functional molecule pepstatin A can occupy the binding site of P-glycoprotein, which helps nanoparticle transport and visualizati...

Embodiment 3

[0055] Stereotactically locate the left hippocampal CA3 area of ​​SD rats, microneedle injection of 3μL kainic acid (KA, 0.5mg / mL) to establish an animal model of epilepsy; injection of 3μL PBS (phosphate buffered saline) as a sham operation control group; 4% poly Perfused with formaldehyde, the brain was taken, and the expression of P-glycoprotein in the hippocampus of the epilepsy group and sham operation group was observed by immunofluorescence confocal imaging and WesternBlot 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 are 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 brain epileptic foci and its preparation and application. Background technique [0002] Epilepsy is one of the most common neurological disorders with chronic, spontaneous, and repetitive clinical features. According to statistics, there are about 9 million patients with epilepsy in my country, and 450,000 new cases are added every year. Although about 1 / 3 of the patients are treated with regular anti-epileptic drugs, their seizures cannot be controlled, which significantly increases the severity of the disease, mortality and Social family economic burden. At present, the effect of epileptic foci resection is relatively positive, but clinical practice shows that the operation is difficult to completely remove, and the disability rate is high, which seriously affects the quality of life of patients. One of the important rea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/12A61K49/14A61K49/18A61K49/00A61K47/04A61K47/42A61K47/48A61K9/51A61P25/08A61K47/60A61K47/62A61K47/69
Inventor 王剑虹张军于向荣耿道颖庞志清潘嘉炜
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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