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Preparation method of double-targeting cationic ultrasound microbubbles carrying cell-penetrating peptide iRGD

An ultrasonic microbubble and cation technology, which is applied in the field of basic research, can solve the problems of low plasmid DNA carried by ultrasonic microbubbles, difficult to achieve microbubble aggregation, and limited number of genes, so as to improve gene transfection efficiency, imaging stability, and echo performance. Good results

Inactive Publication Date: 2015-12-02
SHENZHEN PEOPLES HOSPITAL
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Problems solved by technology

[0004] However, compared with other gene delivery systems, ultrasonic microbubbles currently have a lower ability to carry plasmid DNA, and it is difficult to achieve the accumulation of microbubbles around tumor cells, so that the number of genes entering tumor cells is limited when given ultrasound irradiation, thus affecting the transfection rate

Method used

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  • Preparation method of double-targeting cationic ultrasound microbubbles carrying cell-penetrating peptide iRGD

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Embodiment

[0042] A method for preparing dual-targeted cationic ultrasonic microbubbles carrying iRGD membrane-penetrating peptides, comprising the following process steps:

[0043] (1) Preparation of DSPE-PEG2000-iRGD

[0044] ① Weighing: Weigh the drug iRGD and DSPE-PEG2000-maleimide according to the molar ratio of 1.5:1;

[0045] ②Dissolution: transfer the drug iRGD and DSPE-PEG2000-maleimide into a vial, add 4ml of double distilled water to dissolve, and blow evenly;

[0046] ③ Reaction: Place the vial containing the drug in ice cubes and stir it magnetically for 24 hours;

[0047] ④ Dialysis: Take a large beaker, fill it with distilled water, transfer the reacted solution into a dialysis bag (Mw: 3500) and place it in a large beaker, keep the large beaker in an ice cube environment, and perform magnetic stirring for dialysis for 48 hours. Change the double distilled water and ice every 4 hours;

[0048] ⑤ Freeze-drying: After the dialysis is completed, transfer the dialyzed solut...

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Abstract

The invention discloses a preparation method of double-targeting cationic ultrasound microbubbles carrying cell-penetrating peptide iRGD and belongs to the field of fundamental research. The preparation method uses materials, including DSPC, DSPE-PEG2000-maleimide, DSPE-PEG2000-Biotin, iRGD peptide, CCR2 antibody, stearic acid, branched-chain polyetherimide-600, N, N'-carbonyldiimidazole, and avidin; by bonding integrin Alpha v Beta3 targeting cell-penetrating peptide iRGD and the CCR2 antibody to surfaces of the microbubbles, double-targeting cationic ultrasonic contrast agent is constructed and has the double functions of carrying plasmid DNA and targeting tumor cells; combining ultrasound-mediated biological effect and iRGD cell-penetrating action, the agent is capable of promoting entry of genes into the tumor cells and improving gene transfection efficiency and is expected to be developed and applied to visual ultrasound-controlled-release genetic treatment of tumors.

Description

Technical field [0001] The invention belongs to the field of basic research, and relates to a preparation method of ultrasonic microbubbles, in particular to a preparation method of double-targeted cationic ultrasonic microbubbles carrying iRGD penetrating peptide. Background technique [0002] Ultrasonic microbubbles (microbubble, MB), the main component of ultrasound contrast agents, have the characteristics of high safety, enhanced ultrasound scattering intensity and rich harmonic signals, and are widely used in various types of disease diagnosis, significantly improving the The sensitivity and specificity of diagnosis, the use of ultrasound contrast agent for ultrasound imaging has become an important part of ultrasound diagnosis. [0003] In recent years, with the development of ultrasonic molecular imaging, the use of ultrasonic microbubbles is no longer limited to the field of imaging. Proper modification of ultrasonic microbubbles through biochemical techniques can c...

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Application Information

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IPC IPC(8): A61K49/22A61K48/00A61K47/42A61K41/00A61P35/00
Inventor 徐金锋刘莹莹魏章洪
Owner SHENZHEN PEOPLES HOSPITAL
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