Ultrasonic diagnosis target reagent for liver cancer and preparation method thereof

A technology for ultrasonic diagnosis and targeting reagents, which is applied in echo/ultrasonic imaging agents, liposome delivery, drug delivery, etc., and can solve the problems of affecting biological activity, reducing targeting function, and inability of targeting ultrasound contrast agents to target tissues. Tight adsorption and other issues to achieve the effect of high targeted biological activity

Inactive Publication Date: 2017-09-22
QIANFOSHAN HOSPITAL OF SHANDONG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, the method of linking specific antibodies or ligands to the surface of microbubbles usually adopts biotinylation of specific antibodies or ligands, and then adding avidin, and then through the linking effect of avidin and biotin to make Specific antibodies or ligands are connected to microbubbles. However, there are certain problems in the above-mentioned connection. Because the biotinylation of antibodies or ligands may affect their biological activity and be affected by the shear force of blood flow, targeting ultrasound contrast agents It cannot be tightly adsorbed to the target tissue, which reduces the targeting function, which is especially evident in the contrast agent of liver cancer

Method used

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  • Ultrasonic diagnosis target reagent for liver cancer and preparation method thereof
  • Ultrasonic diagnosis target reagent for liver cancer and preparation method thereof
  • Ultrasonic diagnosis target reagent for liver cancer and preparation method thereof

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preparation example Construction

[0037] In yet another typical embodiment of the present invention, a method for preparing a targeting reagent for ultrasound diagnosis of liver cancer comprises the following steps:

[0038] 1) Dissolve dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE) and polyethylene glycol 1500 in chloroform at a mass ratio of 1:4 to 7:3 to make a suspension, and dissolve the Mix the above suspension, and form a phospholipid film under the action of dry nitrogen;

[0039] 2) Add Tris buffer solution to the phospholipid film, shake the shaker at room temperature to disperse the phospholipid solution to a transparent lipid shell;

[0040] 3) placing the phospholipid solution containing the lipid shell in a vial, replacing the air in the vial with a biologically inert gas, and vibrating to prepare ultrasonic microbubbles;

[0041] 4) Dissolve the Endoglin monoclonal antibody in PBS-EDTA buffer (PE, 4×, pH7.4), dissolve mercaptoethanol in PBS buffer (pH7.4),...

Embodiment

[0056] Experimental procedure

[0057] 1. Animal model BALB / c nude mice, 3-4 weeks old, male, body weight 18-22g, raised in SPF environment, take logarithmic phase HepG2 liver cancer cells, prepare single cell suspension, adjust the cell concentration to 1× 10 7 / ml, subcutaneously inoculate 0.1ml of the suspension in the abdomen of each nude mouse, and perform imaging observation three weeks later;

[0058] 2. Ultrasonic instrument parameter setting adopts the second harmonic imaging mode, the imaging frequency is 10MHz, the mechanical index (MI) is 0.5, and the gain is -20dB;

[0059] 3. Using conventional immunohistochemistry, qRT-PCR, Western blot to analyze the liver tissue of nude mice and transplanted liver cancer tissue; use HepG2 liver cancer cell culture medium to culture human umbilical vein endothelial cells (HUVECs).

[0060] Experimental results

[0061] targeted ultrasound imaging

[0062] All experimental animals received ultrasound contrast imaging, and no...

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Abstract

The invention discloses an ultrasonic diagnosis target reagent for liver cancer. The reagent is an Endoglin target microbubble, the target microbubble comprises a lipid casing and a gas inner core, and an Endoglin monoclonal antibody is connected to the outer surface of the lipid casing. The lipid casing is prepared from dipalmitoyl phosphoethanolamine (DPPE), distearoyl phosphoethanolamine (DSPE) and polyethylene glycol 1500 which are mixed, and the gas inner core is biological inert gas and further optimized as perfluoropropane gas (C3F8). According to the designed ultrasonic diagnosis target reagent disclosed by the invention, the target microbubble can penetrate through tumor vessels through intravenous injection to enter tumor tissue gaps and be specifically combined with adult vascular endothelial cells and mesenchymal cells; thus, the microbubble can effectively enrich at the tumor position, retention of the microbubble is prolonged, lesion tissues and peripheral normal tissues can be clearly distinguished through gathering imaging, and a non-linear harmonic imaging technology is utilized to achieve target molecular imaging of the liver cancer.

Description

technical field [0001] The invention relates to a targeting reagent for ultrasound diagnosis, in particular to a targeting reagent for ultrasound diagnosis of liver cancer and a preparation method thereof. Background technique [0002] Liver cancer is an aggressive cancer that causes high mortality in Asia, and early diagnosis and treatment are important to improve the survival chances of liver cancer patients. Ultrasound examination is an important method for clinical diagnosis of tumors due to its advantages of non-invasive and radiation-free, convenient operation, low cost, and low requirements for equipment and space. However, due to the limitations of ultrasonographic diagnosis itself, it is difficult to detect small liver cancer tissues. [0003] With the emergence of various new ultrasound contrast agents and the continuous development of ultrasound imaging technology, microbubble contrast agents (such as Sonovue contrast agent produced by Braco Company in China) have...

Claims

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Application Information

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IPC IPC(8): A61K49/22
CPCA61K49/227
Inventor 孙洪军单容王蓓刘炬王爱光刘静
Owner QIANFOSHAN HOSPITAL OF SHANDONG
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