CN-121971658-A - Imaging method for displaying brain disease death neuron based on polyethylene glycol modified nano-contrast agent

CN121971658ACN 121971658 ACN121971658 ACN 121971658ACN-121971658-A

Abstract

The invention discloses an imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent, which belongs to the technical field of biomedical imaging. After being injected into the brain disease model animal body by intravenous injection, the brain imaging is performed by using an in-vivo imaging device. The result shows that the contrast agent can cross the damaged blood brain barrier and specifically gather at dead neurons in a focus area, and accurate positioning and visualization of a cell molecular layer can be realized by detecting enrichment signals. The invention breaks through the limitation of the traditional anatomical imaging, and provides a brand-new technical means for early diagnosis, treatment time window judgment and curative effect evaluation of brain injury, neurodegenerative diseases and the like.

Inventors

ZHANG KUN
GAO MINGYUAN
Jiao Mingxia
ZHANG PEISEN
ZENG JIANFENG
WANG JINGYUN

Assignees

苏州欣影生物医药技术有限公司

Dates

Publication Date: 20260505
Application Date: 20260129

Claims (10)

1. An imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent, which is characterized by comprising the following steps: S1, preparing a polyethylene glycol modified nano contrast agent; s2, injecting the polyethylene glycol modified nano-contrast agent into a body of a to-be-detected object in an intravenous injection mode; s3, imaging the brain of the injected object to be detected by using in-vivo imaging equipment; S4, analyzing an imaging result, wherein a specific aggregation area of the polyethylene glycol modified nano-contrast agent in brain tissue indicates the position of dead neurons, so that brain disease diagnosis at a cell or molecular level is realized.
2. The imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent according to claim 1, wherein the preparation method of the polyethylene glycol modified nano-contrast agent in step S1 is selected from any one of the following methods (1), (2) or (3): (1) A preparation method based on an iron oxide nano material comprises the following steps: ① Dissolving organic ferric salt, oleylamine and polyethylene glycol in diphenyl ether or toluene to form a mixed reaction solution, removing oxygen, heating and reacting for 0.5-4 hours to form a nanomaterial, cooling the reaction solution to room temperature, precipitating, centrifuging and washing to obtain the polyethylene glycol modified nano contrast agent; ② Dissolving organic ferric salt and oil-soluble micromolecules in an organic solvent to form a mixed reaction solution, removing oxygen, heating and reacting for 0.5-4 hours to form a nano material, cooling the reaction solution to room temperature, precipitating, centrifuging and washing to obtain a solid nano material, dispersing the solid nano material in tetrahydrofuran, adding polyethylene glycol for surface modification reaction, and purifying after the reaction to obtain the polyethylene glycol modified nano contrast agent; (2) A preparation method based on semiconductor nano-materials comprises the following steps: ① The synthesis of the nano-cores comprises the steps of dissolving inorganic metal salt compounds and water-soluble micromolecules or macromolecules in water to form a mixed reaction solution, adding a first sulfur source or a first selenium source after removing oxygen, and heating and reacting for 0.5-5 hours to form the nano-cores; ② Cooling the reaction solution obtained in the step ① to 70-90 ℃, adding metal inorganic salt for cladding and a second sulfur source or a second selenium source, and continuing to react for 0.5-5 hours to form a shell layer on the surface of the nano core to obtain the nano material with the core-shell structure; ③ Cooling the reaction solution obtained in the step ② to room temperature, precipitating, centrifuging and washing to obtain a solid nano material, dispersing the solid nano material in water, adding polyethylene glycol aqueous dispersion to perform surface modification reaction, and purifying after the reaction to obtain the polyethylene glycol modified nano contrast agent; (3) A preparation method based on rare earth nano materials comprises the following steps: Dissolving an inorganic metal salt compound and oil-soluble micromolecules in an organic solvent to form a mixed reaction solution, removing oxygen, adding a fluorine source, heating and reacting for 0.5-4 hours to form a nano material, cooling the reaction solution to room temperature, precipitating, centrifuging and washing to obtain a solid nano material, dispersing the solid nano material in tetrahydrofuran, adding polyethylene glycol for surface modification reaction, and purifying after the reaction to obtain the polyethylene glycol modified nano contrast agent.
3. The method for imaging neurons showing brain disease death based on polyethylene glycol modified nano-contrast agent according to claim 2, wherein, In the method (1), the organic ferric salt is ferric acetylacetonate or ferric oleate with the concentration of 0.01-100 mmol/L, and the oil-soluble micromolecule is one or more of oleic acid, oleylamine and stearyl alcohol with the concentration of 0.1-1000 mmol/L; in the method (2), the inorganic metal salt compound is a soluble salt of group IIB, group IIIA, group IVA or transition metal, the concentration is 0.01-100 mmol/L, the water-soluble small molecule is an organic small molecule containing carboxyl or sulfhydryl, the high molecule is a water-soluble high polymer, and the concentration is 0.1-1000 mmol/L; The first sulfur source is selected from one or more of sodium sulfide, thiourea or thioacetamide, the first selenium source is selected from one or more of sodium selenate, sodium selenite or selenouride, the concentration is 0.01-600 mmol/L, the metal inorganic salt for cladding is soluble salt selected from zinc, manganese, silver or gold, the concentration is 0.01-100 mmol/L, the second sulfur source is selected from one or more of sodium sulfide, thiourea or glutathione, the second selenium source is selected from one or more of sodium selenate, sodium selenite or glutathione, and the concentration is 0.1-600 mmol/L; In the method (3), the inorganic metal salt compound is selected from one or more of gadolinium chloride, erbium for greening, erbium acetate, thulium chloride, ytterbium acetate and yttrium acetate, and the oil-soluble small molecule is one or more of octadecene and oleic acid; in the methods (1), (2) and (3), the mass ratio of the solid nanomaterial to the polyethylene glycol is 1:10-50.
4. The imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent according to claim 1, wherein the surface of the polyethylene glycol modified nano-contrast agent prepared in step S1 is not modified with any functional molecule targeting death neurons.
5. The imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent according to claim 1, wherein in step S2, the polyethylene glycol modified nano-contrast agent is dispersed in physiological saline to prepare a solution with a concentration of 0.1-20 mg/mL, and the solution is injected into a mouse body through tail vein by 50-200 microliters.
6. The imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent according to claim 1, wherein in step S3, the in vivo imaging device is an optical imaging device, a magnetic resonance imaging device or a computed tomography device.
7. The imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent according to claim 6, wherein when the in vivo imaging device is an optical imaging device, the polyethylene glycol modified nano-contrast agent is a fluorescent quantum dot or a rare earth nanomaterial, and three-dimensional optical imaging of brain tissue is performed in step S3; when the in-vivo imaging device is a magnetic resonance imaging device, the polyethylene glycol modified nano-contrast agent is nano-ferric oxide, and in step S3, brain tissue magnetic resonance imaging is performed.
8. A polyethylene glycol modified nano-contrast agent, wherein the polyethylene glycol modified nano-contrast agent is prepared by the preparation method of the polyethylene glycol modified nano-contrast agent according to claim 2 or 3.
9. Use of a polyethylene glycol modified nano-contrast agent for the preparation of a diagnostic formulation for imaging dead neurons of brain diseases, wherein the diagnostic formulation is capable of specifically aggregating dead neuronal regions in brain tissue after intravenous injection.
10. A kit for carrying out the imaging method according to any one of claims 1 to 7, comprising a dose of polyethylene glycol modified nano-contrast agent according to claim 8 capable of effecting imaging of brain-disease-dead neurons, together with a physiological saline diluent and instructions for use.

Description

Imaging method for displaying brain disease death neuron based on polyethylene glycol modified nano-contrast agent Technical Field The invention relates to the technical field of biomedical imaging, in particular to an imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent. Background Brain diseases, including brain injury, neurodegenerative diseases, and the like, are one of the major health challenges worldwide leading to high mortality and long-term disability. Currently, computed Tomography (CT) and Magnetic Resonance Imaging (MRI) are routine techniques for clinically diagnosing brain lesions. CT is critical for identifying acute hematomas and skull fractures, but has limited ability to visualize the typical features of many brain diseases, such as diffuse axonal lesions, microstructure soft tissue lesions, and the like. MRI, while having superior soft tissue contrast, is more sensitive than CT in detecting microstructural lesions, it essentially provides mainly anatomical information. However, the pathological course of brain disease begins with subtle changes at the cellular and molecular level, much earlier than changes in macroscopic anatomy. The existing clinical imaging modalities are not sufficiently sensitive to these critical cellular and molecular events (e.g., death of specific neurons) at the earliest stage of the lesion, resulting in failure to perform early interventions (e.g., administration of neuroprotective agents) within a critical treatment window, often missing the optimal treatment opportunity, resulting in poor prognosis. In recent years, nano-contrast agents offer new possibilities for high-precision imaging due to their tunable size and surface chemistry. Research has explored the use of various nanomaterials for anatomical localization of brain lesions by MRI or optical imaging. Polyethylene glycol modified nanocontrast agents have also been widely used in the imaging arts to improve biocompatibility and cycle time. However, in brain disease diagnosis, how to achieve accurate targeting and imaging of specific cell types (such as dead neurons) in the disease microenvironment with polyethylene glycol modified nano-contrast agents is still unclear in the interaction mechanism, and the specific diagnostic capability of the cell/molecule level is still limited. Therefore, in order to solve the problems, the invention provides an imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent, which can specifically gather in death neuron areas even though an unmodified targeting molecule is synthesized, and detection is carried out by using in-vivo imaging equipment, so that early accurate diagnosis of brain disease in a cell/molecule layer is realized, and the limitations of traditional anatomical imaging are overcome. Disclosure of Invention The invention aims to provide an imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent, which can specifically display brain disease death neurons and realize accurate diagnosis on a cell/molecular level. The aim of the invention is achieved by the following technical scheme: An imaging method for displaying brain disease death neurons based on polyethylene glycol modified nano-contrast agent, comprising the following steps: S1, preparing a polyethylene glycol modified nano contrast agent; s2, injecting the polyethylene glycol modified nano-contrast agent into a body of a to-be-detected object in an intravenous injection mode; s3, imaging the brain focus of the object to be detected after injection by using in-vivo imaging equipment; S4, analyzing an imaging result, wherein a specific aggregation area of the polyethylene glycol modified nano-contrast agent in brain tissue indicates the position of dead neurons, so that brain disease diagnosis at a cell or molecular level is realized. Preferably, the preparation method of the polyethylene glycol modified nano-contrast agent in the step S1 is selected from any one of the following methods (1), (2) or (3): (1) A preparation method based on an iron oxide nano material comprises the following steps: ① Dissolving organic ferric salt, oleylamine and polyethylene glycol in diphenyl ether or toluene to form a mixed reaction solution, removing oxygen, heating and reacting for 0.5-4 hours to form a nanomaterial, cooling the reaction solution to room temperature, precipitating, centrifuging and washing to obtain the polyethylene glycol modified nano contrast agent; ② Dissolving organic ferric salt and oil-soluble micromolecules in an organic solvent to form a mixed reaction solution, removing oxygen, heating and reacting for 0.5-4 hours to form a nano material, cooling the reaction solution to room temperature, precipitating, centrifuging and washing to obtain a solid nano material, dispersing the solid nano material in tetrahydrofu