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Recombinant human endostatin liposome microbubble and application thereof in preparing medicament for inhibiting tumor angiogenesis under ultrasound mediation

A vascular endothelium and lipid microbubble technology, which can be used in antitumor drugs, echo/ultrasound imaging agents, drug combinations, etc. Anti-tumor angiogenesis effect, significantly inhibiting tumor growth effect

Inactive Publication Date: 2014-03-19
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The currently developed drug-loaded ultrasonic microbubbles are mostly in the micron level, and cannot directly deliver drugs to tumor cells through the vascular endothelial space, making it difficult to achieve true tumor-targeted therapy

Method used

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  • Recombinant human endostatin liposome microbubble and application thereof in preparing medicament for inhibiting tumor angiogenesis under ultrasound mediation
  • Recombinant human endostatin liposome microbubble and application thereof in preparing medicament for inhibiting tumor angiogenesis under ultrasound mediation
  • Recombinant human endostatin liposome microbubble and application thereof in preparing medicament for inhibiting tumor angiogenesis under ultrasound mediation

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Embodiment

[0019] 1. Preparation of Biotinylated Recombinant Human Endostatin

[0020] Biotin-N-hydroxysuccinimide ester (BNHS-ester) is a product of Sigma Company in the United States, 250 mg, molecular weight 341.38, and is prepared into a 1 mg / mL solution with dimethyl sulfoxide. Dissolve BNHS-ester and recombinant human endostatin at a molar ratio of 15:3 in carbonate buffer solution at pH 8.5, incubate at 4°C for 24 hours, add an equal volume of glutamic acid after 24 hours to terminate the reaction, and the reaction solution is supercharged at 10,000 rpm. Filter and centrifuge for 20 min, wash 3 times to obtain biotinylated recombinant human endostatin.

[0021] 2. Preparation of rh-ES-carrying lipid microvesicles

[0022] Disperse DSPE-PEG2000-Biotin and DSPC at a molar ratio of 1:1 in 1 ml of pH 7.40 phosphate buffer; pass inert gas SF6, mechanically shake for 40s; add avidin (avidin: DSPE-PEG2000- Biotin molar ratio is 1:2), incubate at room temperature for 1 h; add biotinylat...

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Abstract

The invention discloses a recombinant human endostatin liposome microbubble, which is prepared by a method in the following steps: dispersing DSPE-PEG (distearoyl phosphoethanolamine-polyethylene glycol) 2000-Biotin and DSPC (disaturated phosphatidylcholine) in a molar ratio of 1:(0.5-4) in a phosphate buffer; introducing inert gas, mechanically oscillating, adding streptavidin, and incubating at room temperature, wherein the molar ratio of the streptavidin to the DSPE-PEG2000-Biotin is 1:(1-2); and adding biotinylated recombinant human endostatin, and incubating at room temperature to prepare the recombinant human endostatin liposome microbubble, wherein the molar ratio of the streptavidin to the biotinylated recombinant human endostatin is 1:(1-2). The invention also discloses application of the recombinant human endostatin liposome microbubble in preparing a medicament for inhibiting tumor angiogenesis under ultrasound mediation. The recombinant human endostatin liposome microbubble has a tumor inhibition rate of 41.3 percent, which is remarkably superior to the inhibition effect of a single recombinant human endostatin liposome microbubble.

Description

technical field [0001] The present invention relates to a new preparation for anti-tumor angiogenesis. Specifically, the present invention prepares a lipid microbubble of recombinant human endostatin, which can cause the drug-loaded microbubble to burst and release the drug in the local tumor under ultrasonic irradiation. And the ultrasound biological effect is used to synergistically inhibit tumor angiogenesis. Background technique [0002] Endostatin specifically acts on neovascular endothelial cells, inhibits their growth and migration, induces apoptosis, and thus inhibits angiogenesis. Recombinant human endostatin (rh-ES) has 9 amino acids added to the N-terminus of endostatin, which improves the purity, stability and biological activity of the protein, and has been used in clinical anti-tumor. However, due to the short plasma half-life of rh-ES, daily intravenous administration is required clinically, which is cumbersome to operate. At the same time, due to the lack o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/22A61K41/00A61K38/17A61K9/00A61K47/34A61P35/00
Inventor 陈志奎林礼务薛恩生张秀娟杨菁
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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