Gene-ordrug-carrying-carrying ultrasonic microvesicle contrast-media and preparing method thereof

An ultrasonic microbubble and contrast agent technology, applied in the direction of MRI/magnetic resonance imaging contrast agent, etc., can solve the problems of inability to ensure gene delivery to target tissues, poor repeatability and stability, and inability to prevent nuclease degradation, etc. Prevent washout and dilution, increase efficacy, and stabilize the effect

Inactive Publication Date: 2006-08-09
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For gene-carrying microbubbles, at this stage, the adhesion of albumin or the surface charge characteristics of lipid microbubbles are mostly used to simply attach genes to the surface of microbubbles to prepare gene-adhered microbubbles. Not only the number of genes it carries Limited, high variability, poor reproducibility and stability, seriously affecting gene transfection efficiency
More importantly, since the gene is exposed on the surface of the microbubble, when the gene is applied in vivo, it cannot prevent the degradation of the gene by nuclease in the blood, so the microbubble cannot guarantee the delivery of a large amount of gene to the target tissue

Method used

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  • Gene-ordrug-carrying-carrying ultrasonic microvesicle contrast-media and preparing method thereof
  • Gene-ordrug-carrying-carrying ultrasonic microvesicle contrast-media and preparing method thereof
  • Gene-ordrug-carrying-carrying ultrasonic microvesicle contrast-media and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] [Example 1] Preparation of ultrasonic microbubble contrast agent

[0073] 1. Weigh raw materials:

[0074] Distearoylphosphatidylcholine DSPC 5mg

[0075] Dipalmitoylphosphatidylethanolamine DPPE 2mg

[0076] span60 1mg

[0077] Glycerol 50μl

[0078] Plasmid PBS 450μl;

[0079] 2. After mixing the above raw materials, place them in a 1.5ml vial containing 100% perfluoropropane;

[0080] 3. Place the vial in a water bath at 40°C for 30 minutes;

[0081] 4. Oscillate the vial with a dental oscillator for 35 seconds;

[0082] 5. Let it stand for 5 minutes, add 0.5ml of PBS to dilute, and then obtain the ultrasonic microbubble contrast agent.

Embodiment 2

[0083] [Example 2] Preparation of ultrasonic microbubble contrast agent loaded with vascular endothelial growth factor VEGF

[0084] 1. Weigh raw materials:

[0085] Distearoylphosphatidylcholine DSPC 5mg

[0086] Dipalmitoylphosphatidylethanolamine DPPE 2mg

[0087] span60 1mg

[0088] Glycerol 50μl

[0089] PBS 450μl

[0090] Vascular endothelial growth factor VEGF 1mg;

[0091] 2. After mixing the above raw materials by shaking method, put them in a 1.5ml vial containing perfluoropropane with a purity of 99.99%;

[0092] 3. Place the vial in a water bath at 40°C for 30 minutes;

[0093] 4. Oscillate the vial with a dental oscillator for 35 seconds;

[0094] 5. Let it stand for 5 minutes, add 0.5ml PBS to dilute, and you can get the gene-carrying ultrasonic microbubble contrast agent.

Embodiment 3

[0095] [Example 3] Preparation of ultrasonic microbubble contrast agent loaded with basic fibroblast growth factor bFGF

[0096] 1. Weigh raw materials by weight percentage:

[0097] Distearoylphosphatidylcholine DSPC 4mg

[0098] Dipalmitoylphosphatidylethanolamine DPPE 2.5mg

[0099] span60 0.9mg

[0100] Glycerol 40μl

[0101] PBS 460μl

[0102] Basic fibroblast growth factor bFGF 2.5mg;

[0103] 2. After mixing the above raw materials by means of oscillation, place them in a 1.5ml vial containing perfluoropropane with a purity of 99%;

[0104] 3. Place the vial in a water bath at 40°C for 30 minutes;

[0105] 4. Oscillate the vial with a dental oscillator for 35 seconds;

[0106] 5. Let it stand for 5 minutes, add 0.5ml of PBS to dilute, and then the gene-loaded ultrasonic microbubble contrast agent can be obtained.

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Abstract

This invention discloses an ultrasonic microbubble contrast agent of carrier gene, it includes microbubble, fluorocarbon gas, gene or medicine. Microbubble wall is made up by lipid bielement, fluorocarbon gas is encapsulated in it, gene or medicine entrapped in microbubble envelope. This invention also discloses its preparation method, two stearyl phosphatidylcholine, two palmitoyl phosphatidyl ethanolamine, sapn-60, glycerol, phosphate buffer solution and gene or medicine is mixed as proper proportion for mechanical oscillations. The property of carry gene or medicine microbubble is stable, particle diameter disperses equally, local tissue medicine concentration can be obviously advanced though effect of transonic breaking microbubble, and curative effect is advanced and side effect can be reduced. Flushing and dilution of blood to gene and medicine can be prevented when used in body, and also prevent gene or medicine from degrading by nuclease in cycle to ensure that lot of gene and medicine can be transported to target tissue. And it is linear increase follow gene or medicine input amount in certain range.

Description

technical field [0001] The invention relates to an ultrasonic microbubble contrast agent capable of efficiently carrying genes or drugs and a preparation method thereof, which provides a new type of high-efficiency delivery carrier that can be injected intravenously for gene or drug targeted therapy, and is not only suitable for ultrasonic rupture of microbubble targets The transfer of genes and drugs to treat diseases of the cardiovascular system can also provide an effective delivery tool for targeted therapy of parenchymal organ diseases. Background technique [0002] 1. Development status of ultrasound contrast agents [0003] Acoustic contrast agent is composed of gas-containing microbubbles similar to the size of red blood cells. After being injected into the vein, it can reach the parts that red blood cells can reach. For example, myocardium, liver, kidney, etc. produce strong contrast when detected by two-dimensional ultrasound equipment. Compared with other imagin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/06A61K49/08
Inventor 王志刚冉海涛李兴升任红凌智瑜张群霞郑元义许川山杨春江
Owner CHONGQING MEDICAL UNIVERSITY
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