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Preparation method of T1 strengthened nuclear magnetic resonance nanometer contrast agent

A nano-contrast agent, nuclear magnetic resonance technology, applied in the preparations, pharmaceutical formulations, emulsion delivery and other directions for in vivo experiments, can solve the problems of poor pharmacokinetics, low sensitivity, poor specificity, etc., and achieve biocompatibility Excellent, good enhancement, accurate visualization

Inactive Publication Date: 2020-01-03
广州市番禺区中心医院
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  • Abstract
  • Description
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Problems solved by technology

Although technetium-99-labeled fibrinogen and iodine-131-labeled anti-fibrin antibody have been used in the diagnosis of thrombosis, their clinical application is limited due to their poor pharmacokinetics, low sensitivity, and poor specificity in vivo (Leung J M, Berry L R, Chan A K C, et al.Surface modification of polydimethylsiloxane with a covalent antithrombin–heparin complex to prevent thrombosis[J].Journal of Biomaterials Science, Polymer Edition,2014,25(8):786-801.)

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  • Preparation method of T1 strengthened nuclear magnetic resonance nanometer contrast agent
  • Preparation method of T1 strengthened nuclear magnetic resonance nanometer contrast agent
  • Preparation method of T1 strengthened nuclear magnetic resonance nanometer contrast agent

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preparation example Construction

[0029] A preparation method of a T1-enhanced nuclear magnetic resonance nano-contrast agent, which includes selecting a Gd contrast agent as a part of the T1-enhanced MRI imaging contrast agent, realizing the dissociation and self-assembly of ferritin by adjusting the pH of the reaction solution, and then transferring the Gd contrast agent to In the ferritin nanocage, the KGDS tetrapeptide which is targeted to the activated platelet membrane glycoprotein GPIIb / IIIa was selected as the targeting ligand, and the KGDS tetrapeptide was coupled to the surface of apoferritin by chemical methods to obtain the The above-mentioned T1-enhanced nuclear magnetic resonance nano-contrast agent with activated platelet targeting.

[0030]The pH adjustment range of the reaction solution is between 2.0-7.4; the molar ratio of ferritin to KGDS tetrapeptide is 1:25-1:100; the nano-contrast agent has good biocompatibility, It has active targeting to activated platelets; the size of the nano-contra...

Embodiment 1

[0047] Step 1: Use 1M hydrochloric acid to adjust the pH value of the deferrified horse spleen ferritin aqueous solution to 2.0, the ferritin concentration is 10mg / mL, stir and incubate for 15min; take 100mg of Gd-DOTA solid, slowly add it to 2mL ferritin solution, and stir While adding, wait until the Gd-DOTA solid is completely dissolved, and mix with the apoferritin solution, stir at room temperature for 20 minutes, adjust the pH of the reaction solution to 7.4 with 1M sodium hydroxide, and continue stirring for 2 hours;

[0048] Step 2: After the reaction, centrifuge the reaction solution obtained in step (1) for 5 minutes at a speed of 12000 rpm / min to remove the protein precipitate produced during the ferritin self-assembly process;

[0049] Step 3: After centrifugation, transfer the supernatant in step (2) to a dialysis bag with a molecular weight cut-off of 8000-12000, put it into PBS buffer solution and dialyze for 24 hours, and change the dialysate every 8 hours to ob...

Embodiment 2

[0053] Step 1: Use 1M hydrochloric acid to adjust the pH value of the deferrified horse spleen ferritin aqueous solution to 2.0, the ferritin concentration is 10mg / mL, stir and incubate for 15min; take 100mg of Gd-DOTA solid, slowly add it to 2mL ferritin solution, and stir While adding, wait until the Gd-DOTA solid is completely dissolved, and mix with the apoferritin solution, stir at room temperature for 20 minutes, adjust the pH of the reaction solution to 7.4 with 1M sodium hydroxide, and continue stirring for 2 hours;

[0054] Step 2: After the reaction, centrifuge the reaction solution obtained in step (1) for 5 minutes at a speed of 12000 rpm / min to remove the protein precipitate produced during the ferritin self-assembly process;

[0055] Step 3: After centrifugation, transfer the supernatant in step (2) to a dialysis bag with a molecular weight cut-off of 8000-12000, put it into PBS buffer solution and dialyze for 24 hours, and change the dialysate every 8 hours to ob...

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Abstract

The invention discloses a T1 strengthened nanometer magnetic contrast agent having the function of activating targeting properties of blood platelets and a preparation method of the T1 strengthened nanometer magnetic contrast agent belonging to the field of nuclear magnetic resonance contrast agents. The characteristics of "dissociation-self-assembly" and surface modificability of an apoferritin nanometer cage under different pH environment are utilized, paramagnetic Gd-DOTA is loaded into the apoferritin nanometer cage, ligand-KGDS tetrapeptide specifically targeting and activating the bloodplatelets is coupled to the surface of Gd-loaded apoferritin, and therefore, nanometer particles which can actively target and activate the blood platelets and have nuclear magnetic resonance T1 weighted imaging strengthened contrast effects can be obtained. The particle diameter of the nanometer contrast agent is about 10nm, the nanometer contrast agent has favorable contrast effects, targeting properties for activating the blood platelets, and biocompatibility; precise visualization of thrombosis is hopefully realized through targeting and activating the blood platelets, and more comprehensive molecule information is provided for early prevention, diagnosis and treatment of cardiovascular diseases.

Description

technical field [0001] The invention belongs to the field of nuclear magnetic resonance contrast agents, in particular to a preparation method of a T1-enhanced nuclear magnetic resonance nano contrast agent. Background technique [0002] Atherosclerotic thrombosis is the acute thrombosis caused by sudden plaque rupture during the slow progression of atherosclerosis and can lead to serious clinical events such as acute coronary syndrome, transient ischemic attack, and stroke . According to statistics, approximately one out of every three deaths is due to atherosclerotic thrombosis (Xie Yang, Rao Bangfu. Current status of antiplatelet therapy for atherosclerotic thrombosis. World Today's Medical Journal. 2005, 6 (1 ):57-59.). Although intravascular ultrasound elastography, angioscopy, angiography, and temperature measurement have all been applied to imaging plaques, their applications are greatly limited due to their invasiveness. Therefore, it is of great significance to f...

Claims

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Application Information

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IPC IPC(8): A61K49/18A61K49/14
CPCA61K49/085A61K49/14A61K49/1866
Inventor 罗文峰李莉郭惠庄陈汉威叶裕丰黄晨
Owner 广州市番禺区中心医院
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