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99Tcm-Cys-AnnexinV labeled compound with phosphatidylserine as target spot and labeling process and application thereof

A technology of 99tcm-cys-annexinv and phosphatidylserine, which is applied in the direction of radioactive physical shape and radioactive carrier, can solve the problems of increased radioactive uptake in the liver and the influence of AnnexinV activity, and achieves rapid liver and kidney clearance, which is conducive to clinical transformation and labeling The effect of the simple and easy process

Inactive Publication Date: 2013-04-17
NANJING FIRST HOSPITAL +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Direct reduction labeling method is more convenient and ideal than bifunctional chelating agent method. Under mild pH conditions, 99Tcm-AnnexinV-labeled products with high labeling rate (>95%) and good stability (>10h) can be obtained, and are easy to kit However, due to the combination of 99Tcm with the sulfhydryl group in AnnexinV, the activity of AnnexinV will be affected, and some denatured proteins will be produced, which will increase the radioactive uptake of the liver

Method used

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  • 99Tcm-Cys-AnnexinV labeled compound with phosphatidylserine as target spot and labeling process and application thereof
  • 99Tcm-Cys-AnnexinV labeled compound with phosphatidylserine as target spot and labeling process and application thereof
  • 99Tcm-Cys-AnnexinV labeled compound with phosphatidylserine as target spot and labeling process and application thereof

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Embodiment 1

[0039] Embodiment 1. Cys-AnnexinV 99 Tc m Marking technology

[0040] Cys-AnnexinV is provided by the State Key Laboratory of Medical Biotechnology of Nanjing University. It uses genetic engineering technology to clone, express and purify in Escherichia coli, and adds a cysteine ​​with a sulfhydryl group to the carboxyl end of Annexin V, and uses a metal affinity layer. Analysis and purification, recombinant preparation of Cys-AnnexinV. It can be prepared by referring to the method disclosed in the Chinese patent application No. 201110072451.2 entitled "Human Annexin V variant and its expression, preparation and application".

[0041] Cys-AnnexinV was detected by direct labeling method 99 Tc m mark. Take an appropriate amount of SnCl 2 2H 2 O, dissolved in HCl with a concentration of 0.05mol / L, and diluted to 1mg / mL with pH7.4 phosphate buffer. Take the prepared SnCl-containing 2 30μL of citric acid buffer solution, add Cys-AnnexinV 5~10μg, shake well, and then add ...

Embodiment 2

[0044] Embodiment 2. 99 Tc m The biodistribution characteristics of the -Cys-AnnexinV-labeled compound meet the requirements for imaging agents, and are illustrated by organ distribution experiments in mice.

[0045] 40 Kunming mice were randomly divided into 8 groups with 5 mice in each group. tail vein injection 99 Tc m -Cys-AnnexinV at a dose of 11.1MBq in a volume of 0.2mL. A group of animals were executed at 6 time points of 5min, 30min, 1h, 2h, 4h, and 6h after administration, and blood, heart, liver, spleen, lung, kidney, stomach, small intestine, brain, muscle, and bone were taken out. Organs were weighed and radioactive counts were determined, and the time radioactive curves of each organ were drawn; the percent injection dose rate per gram of tissue was calculated (%ID / g, radioactive count per gram of tissue or organ / total radioactive count injected into mice ×100%). The blood clearance curve was drawn with time as the abscissa and radioactive count as the ordi...

Embodiment 3

[0049] Embodiment 3. 99 Tc m -Application of a Cys-AnnexinV labeled compound as an imaging agent for detecting cell apoptosis.

[0050] The technical scheme of the present invention is 99 Tc m -Application of Cys-AnnexinV labeled compound as imaging agent for detecting cell apoptosis. The aforementioned apoptosis can theoretically be the apoptosis process of any normal or diseased tissue cells in the organism, such as cardiomyocytes, tumor cells, etc. In the following, the cardiomyocyte apoptosis, tumor cell apoptosis and platelet activation caused by ischemia-reperfusion-acute myocardial infarction and tumor chemotherapy are taken as examples to illustrate.

[0051] 1. 99 Tc m -Cys-AnnexinV labeled compound as imaging agent to detect cardiomyocyte apoptosis

[0052] Six experimental minipigs (provided by the Experimental Animal Center of Nanjing Military General Hospital), weighing 15-20 Kg, were anesthetized by intramuscular injection of 10 mg / Kg of ketamine hydrochlo...

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Abstract

The invention provides a radio pharmaceutical 99Tcm-Cys-AnnexinV with phosphatidylserine as a target spot, wherein a c-terminus of AnnexinV is connected with cysteine Cys, and radio nuclide 99Tcm is labeled on thiol of the cysteine. Cys-AnnexinV is subjected to 99Tcm label by utilizing a direct labeling method, and a label process comprises the following steps: (1) taking a suitable amount of SnCl2, adding the Cys-AnnexinV to tin protein; and (2) adding 99TcmO4- to the Cys-AnnexinV with tined protein, uniformly mixing and standing to form the 99Tcm-Cys-AnnexinV. According to the invention, the labeling method is more simple and practical, the activity and biologic functions of labeled protein can be kept to the maximum extent, and the label method is applied to image radio nuclide, can be used for detecting the degree and range of apoptosis in various diseases, is especially applied to detecting the apoptosis and the thrombus when serving as an imaging agent.

Description

technical field [0001] The invention belongs to the technical field of radionuclide labeling, in particular to a 99 Tc m -Cys-AnnexinV labeled compound and its labeling technology, and its application as an imaging agent in acute myocardial injury, tumor efficacy evaluation, and acute thrombus detection. Background technique [0002] Apoptosis is a highly conserved programmed cell death (PCD), which plays an important role in many physiological and pathological processes. Excessive apoptosis can lead to loss of tissue function, such as acute myocardial infarction, chronic heart failure, transplant rejection, stroke, neurodegenerative diseases, etc. In contrast, systemic lupus erythematosus and rheumatoid arthritis are characterized by insufficient apoptosis, which allows a large number of immune cells to survive and inappropriately kill and destroy normal tissues, resulting in impaired tissue and organ functions. Inhibition of apoptosis leads to excessive cell proliferati...

Claims

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Application Information

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IPC IPC(8): A61K51/08A61K51/12A61K103/10
Inventor 王峰华子春胡敏进方纬王自正
Owner NANJING FIRST HOSPITAL