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Ultra-small functional nanocluster, nanoprobe and preparation method and application of nanoprobe

A nanoprobe and functionalization technology, applied in the field of nanomedicine, can solve problems such as low targeting efficiency, affecting imaging effect, and increasing particle size of nanomaterials, so as to improve targeting ability, high production yield, and low toxicity Effect

Pending Publication Date: 2018-07-13
THE SECOND HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the method of making nanomaterials have multiple imaging modalities is mainly to assemble different imaging elements, but usually the particle size of nanomaterials increases to a large extent, and they are easily absorbed by the endothelial network after entering the blood circulation system in the body. The system captures most of the nanomaterials into the liver, spleen and other tissues, resulting in only a small amount of nanomaterials reaching the tumor-targeted area. The targeting efficiency is low, which affects the proper imaging effect

Method used

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  • Ultra-small functional nanocluster, nanoprobe and preparation method and application of nanoprobe
  • Ultra-small functional nanocluster, nanoprobe and preparation method and application of nanoprobe
  • Ultra-small functional nanocluster, nanoprobe and preparation method and application of nanoprobe

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Put 2ml of chloroauric acid solution (concentration: 24mM), 1ml of gadolinium chloride solution (48mM), and 24mL of glutathione solution (4mM) in a 100mL single-necked flask, and stir slowly (1000rpm ) after 20 min, the temperature was raised to 70° C. to form nanoclusters, and after 24 h of reaction, the supernatant was removed by centrifugation for 20 min, and washed three times with 10 mL deionized water to obtain nanoclusters.

[0025] 30 μL of nanoclusters (50 mg / mL), 10 μL of EDC (8 mg / mL) and 20 μL of NHS (8 mg) were mixed in 1 mL of phosphate buffer (PBS buffer, 1×, pH 7.4), shaken at 37°C for 20 minutes, and then added 5 μL of cNGR polypeptide (1 mg / mL) was reacted for 2 hours, purified by centrifugation and stored in PBS buffer (1×, pH 7.4) to obtain ultra-small functionalized nanoprobes.

[0026] The obtained nanoprobe was diluted to 0.5 mg / mL, and its ultraviolet absorption spectrum was detected by a UV-visible spectrophotometer, its particle size was detect...

Embodiment 2

[0031] Put 2ml of chloroauric acid solution (concentration: 24mM), 1ml of gadolinium chloride solution (120mM), 24mL of glutathione solution (4mM) in a 100mL single-necked flask, stir slowly (1000rpm) under the condition of water bath (30°C) ) After 20 min, the temperature was raised to 70 °C to form nanoclusters. After 24 hours of reaction, the supernatant was removed by centrifugation for 20 minutes, and washed three times with 10 mL of deionized water to obtain nanoclusters.

[0032] 30 μL of nanoclusters (50 mg / mL), 10 μL of EDC (8 mg / mL) and 20 μL of NHS (8 mg) were mixed in 1 mL of phosphate buffer (PBS buffer, 1×, pH 7.4), shaken at 37°C for 20 minutes, and then added 10 μL of cNGR polypeptide (1 mg / mL), continued to react for 2 h, purified by centrifugation and stored in PBS buffer (1×, pH 7.4). The particle size of the prepared nanoprobe is 6nm.

Embodiment example 3

[0034] Put 2ml of chloroauric acid solution (concentration: 24mM), 1ml of gadolinium chloride solution (120mM), 24mL of glutathione solution (5mM) in a 100mL single-necked flask, and stir slowly (1000rpm ) After 20 min, the temperature was raised to 70 °C to form nanoclusters. After 24 hours of reaction, the supernatant was removed by centrifugation for 20 minutes, and washed three times with 10 mL of deionized water to obtain nanoclusters.

[0035]30 μL of nanoclusters (50 mg / mL), 10 μL of EDC (8 mg / mL) and 20 μL of NHS (8 mg) were mixed in 1 mL of phosphate buffer (PBS buffer, 1×, pH 7.4), shaken at 37°C for 20 minutes, and then added 15 μL of cNGR polypeptide (1 mg / mL), continued to react for 2 h, purified by centrifugation and stored in PBS buffer (1×, pH 7.4). The particle size of the prepared nanoprobe is 3.8nm.

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Abstract

The invention provides an ultra-small functional nanocluster. A nanoculster structure is formed by taking GSH as a template and chelating sulfydryl and carboxyl on the surface of GSH with Au atom andgadolinium Gd atom; two types of atoms are simultaneously coated in the nanoculster structure. The invention also provides an ultra-small functional nanoprobe with the surface modified by cNGR and a preparation method thereof. The ultra-small functional nanoprobe has the capability of specifically identifying tumor angiogenesis; the quantum confinement effect of the Au atom is used for generatingfluorescence and absorbing X-ray according to the adsorption coefficient, so that fluorescence imaging and CT imaging can be achieved; the Gd atom is used for shortening longitudinal relaxation time of free water protons and enhancing MR contrast ratio, so that the application of the ultra-small functional nanoprobe in the multimodal imaging of the tumor angiogenesis can be achieved. The preparednanoparticles are small in size being smaller than 10nm, high in biocompatibility and low in toxicity; through the application of modular assembly performance of the nano material, efficient synergy of multiple functions is achieved; the ultra-small functional nanoprobe is high in preparation yield and is suitable for large-batch production.

Description

technical field [0001] The invention relates to the field of nano-medicine, in particular to an ultra-small functionalized nano-cluster, a nano-probe and a preparation method and application thereof. Background technique [0002] The morbidity and mortality of malignant tumors are increasing year by year. Early diagnosis and personalized treatment of tumors are the most effective ways to improve the survival rate of patients. However, most patients have reached the middle and advanced stages when they are diagnosed, and the tumor has already metastasized, so they miss the best time for treatment. Accurate assessment of tumor microvessels has important clinical significance for evaluating tumor progression, formulating individualized treatment plans, and predicting prognosis. However, the existing clinical contrast agents (gadopentetate sodium, iohexol, etc.) lack targeting and poor sensitivity to directly image living microvessels with diameters less than 300 μm, making it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/18A61K49/14A61K49/04A61K49/00
CPCA61K49/0002A61K49/0056A61K49/0093A61K49/04A61K49/14A61K49/1866
Inventor 李雪张雪宁吴梦琳郭琪张燕燕王嘉慧李亮
Owner THE SECOND HOSPITAL OF TIANJIN MEDICAL UNIV
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