Application of chylomicron as liver targeting gene treatment vector

A chylomicron, gene therapy technology, applied in gene therapy, non-effective components of polymer compounds, genetic material components, etc., can solve the problems of large cytotoxicity and weak carrier specificity, etc.

Active Publication Date: 2011-09-28
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the high positive charge density makes PEI exhibit great cytotoxicity at the same time, and the specificity of the carrier is not strong

Method used

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  • Application of chylomicron as liver targeting gene treatment vector
  • Application of chylomicron as liver targeting gene treatment vector
  • Application of chylomicron as liver targeting gene treatment vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation of a dual-targeting gene vector in which chylomicrons encapsulate the pAFP-TK-IRES2-EGFP plasmid vector.

[0024] Prepare the pAFP-TK-IRES2-EGFP plasmid vector with a concentration of 50 μg / mL, place the chylomicron in a sonicator and sonicate 10 times, each time for 15 s, with an ultrasonic power of 150 W, according to the mass volume ratio of the plasmid vector to chylomicron 2 μg / mL The plasmid vector and chylomicron were mixed, vortexed for 30 seconds, and left to stand for 30 minutes to prepare a dual-targeting gene carrier in which the chylomicron encapsulated the pAFP-TK-IRES2-EGFP plasmid vector.

[0025] The eukaryotic expression plasmid vector pAFP-TK-IRES2-EGFP was purchased from Beijing Xiercheng Industrial Technology Co., Ltd., and the plasmid map of the plasmid vector is as follows: figure 1 , which contains the alpha-fetoprotein (AFP) promoter for gene expression, the AFP promoter can only be activated under a specific AFP activatio...

Embodiment 2

[0026] Example 2: Encapsulation effect of chylomicron on pAFP-TK-IRES2-EGFP plasmid vector.

[0027] The dual-targeting carrier obtained in Example 1 was observed by scanning electron microscopy, and the 100,000-fold scanning electron microscope image showed that chylomicrons had a good encapsulation effect on the pAFP-TK-IRES2-EGFP plasmid vector, and the particle size was between 200 and 300 nm. ,Such as figure 2 .

Embodiment 3

[0028] Example 3: Protective effect of chylomicron on pAFP-TK-IRES2-EGFP plasmid vector.

[0029] Bovine pancreatic deoxyribonuclease I was added to the dual-targeting vector obtained in Example 1 to make the final concentration 20 μg / mL, and digested at 37°C for 30 minutes. The simple pAFP-TK-IRES2-EGFP plasmid vector was used as a control. The protective effect of chylomicron on pAFP-TK-IRES2-EGFP plasmid vector was observed by agarose gel electrophoresis. Observation see image 3 , Lane 1 shows that the simple pAFP-TK-IRES2-EGFP plasmid vector is degraded by bovine pancreatic deoxyribonuclease Ⅰ because there is no protective effect of chylomicrons; Lane 2 shows the effect of chylomicrons on pAFP-TK-IRES2 The -EGFP plasmid vector has a strong protective effect and can avoid its degradation by bovine pancreatic deoxyribonuclease Ⅰ. The results showed that the chylomicrons in the dual-targeting vector had a strong protective effect on the pAFP-TK-IRES2-EGFP plasmid vector a...

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Abstract

The invention relates to new application of chylomicron as a vector of a gene treatment medicament, in particular to application of chylomicron as a vector of a liver targeting gene treatment medicament. Apolipoprotein E (apo E) is contained in the chylomicron, and apo E receptors present on membrane surfaces of liver cells and liver cancer cells, and can identity and combine the apo E specifically, so a plasmid vector can be carried by the chylomicron specifically to be oriented to the liver cells and the liver cancer cells without being absorbed by other cells of bodies; and the chylomicronhas high liver cell targeting, and can be metabolized in the liver cells and the liver cancer cells without residues, so the problem of cytotoxicity is solved.

Description

technical field [0001] The present invention relates to the application of chylomicron, in particular to a brand-new application of chylomicron as a gene therapy carrier. Background technique [0002] Chylomicron (CM) is a reprocessed lipid droplet synthesized by small intestinal mucosal epithelial cells with a diameter of 80-500 nm, containing triglyceride, cholesterol ester and some apolipoproteins such as apoAI, apoAII, apoAIV, apoB 48 , apoCⅠ, apoCⅡ, apoCⅢ, apoE, etc., whose molecular weight is greater than 50×10 6 , the density is less than 0.95g / cm 3 , the main function is to transport exogenous triglycerides to skeletal muscle, cardiac muscle, fat and other tissues, and transport exogenous cholesterol to the liver. [0003] Lipids in food are re-esterified on the smooth endoplasmic reticulum of cells and apolipoprotein synthesized on the rough endoplasmic reticulum to form nascent chylomicrons, which are secreted out of the cell through the Golgi complex, enter the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/42A61P35/00
Inventor 解军于保锋徐钧司海东张小曼程凯王惠珍张悦红赵虹刘志贞张俊涛弓韬胡晓年向前
Owner SHANXI MEDICAL UNIV
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