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Brain-glioma-targeting therapy gene delivery vector and delivery system

A technology for gene delivery and brain glioma, applied in gene therapy, antineoplastic drugs, drug delivery, etc., can solve the problem that the therapeutic gene needs to be improved, and achieve the effect of improving efficiency and targeting efficiency

Active Publication Date: 2020-12-18
BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the application of the above-mentioned technologies still needs to be improved to improve the targeted penetration ability of therapeutic genes.

Method used

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  • Brain-glioma-targeting therapy gene delivery vector and delivery system
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  • Brain-glioma-targeting therapy gene delivery vector and delivery system

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Experimental program
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preparation example Construction

[0048] The preparation method of the therapeutic gene delivery system of the present application is illustrated as follows, wherein:

[0049] The reduction-sensitive amphiphilic copolymer is PCL-ss-PEG-ss-PCL, the functionalized phospholipid is DSPE-PEG-Mal with active groups, the cationic lipid is DOTAP, the active brain targeting molecule is Ang, and the cell penetration The membrane peptide is TAT. Therapeutic genes can be self-extracted, amplified or provided by commercial companies.

[0050] Preparation methods include:

[0051] PCL-ss-PEG-ss-PCL, DSPE-PEG-Mal, DOTAP and Fe 3 o 4 Add it into the eggplant-shaped bottle with a molar ratio of (1-20):1:(1-20):(10-30), add 5mL of dichloromethane, mix well, and install the eggplant-shaped bottle on the rotary evaporator On the top, the bottom of the bottle is in contact with the water bath device, heated to 37°C, and coated by rotary evaporation at 120rpm and 580kPa for 1 hour. After the liquid in the bottle is completely e...

Embodiment 1

[0059] (1) 20mg PCL 3750 -ss-PEG 7500 -ss-PCL3750, 0.8mg DSPE-PEG 2000 -Mal, 1.02mg DOTAP and 1.0mg oleic acid modified Fe 3 o 4 (Molar ratio: 5:1:5:15, PCL 3750 -ss-PEG 7500 -ss-PCL 3750 The molecular weight is 15000, DSPE-PEG 2000 -Mal has a molecular weight of 2893, where PCL 3750 -ss-PEG 7500 -ss-PCL 3750 Synthesized according to conventional methods; DSPE-PEG 2000 -Mal and DOTAP were purchased from Nanocs Biological Technology (Beijing, China); oleic acid-modified Fe 3 o 4 The magnetic nanoparticles were synthesized by Xi'an Ruixi Biotechnology Co., Ltd. (R-CY1010, particle size 10nm) was added to a 5mL eggplant-shaped bottle, 5mL of dichloromethane was added, mixed evenly, and the eggplant-shaped bottle was installed on a rotary evaporator , the bottom of the bottle is in contact with the water bath device, heated to 37°C, and applied by rotary evaporation at 120rpm and 580kPa for 1h. After the liquid in the bottle is completely evaporated, adjust the rotary ev...

Embodiment 2

[0073] Comparison of particle size, potential, stability and cytotoxicity of different modified phospholipid hybrid polymer nanomicelles

[0074] Add 20mg of PCL-ss-PEG-ss-PCL, 0.8mg of DSPE-PEG-Mal, and 1.02mg of DOTAP into a 5mL eggplant-shaped bottle, add 5mL of dichloromethane, mix well, and install the eggplant-shaped bottle on a rotary evaporator , the bottom of the bottle is in contact with the water bath device, heated to 37°C, and applied by rotary evaporation at 120rpm and 580kPa for 1h. After the liquid in the bottle is completely evaporated, adjust the rotary evaporator to vacuum and continue to dry for 1h. Dry overnight in a desiccator. After overnight, 5 mL of HEPES buffer was added and hydrated at 65 °C for 5 h. After returning to room temperature, ultrasound was performed in an ice bath for 10 minutes (1-2 minutes rest every 5 minutes of ultrasound, 5 mm probe, Ampl 30%). The sonicated carrier was dialyzed with PBS for 2 h, during which the dialysate was chan...

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Abstract

The invention provides a brain-glioma-targeting therapy gene delivery vector and a delivery system. The therapy gene delivery vector comprises targeting molecules, wherein the targeting molecules comprise active-brain-targeting molecules and cell-penetrating peptides; a vector skeleton, wherein the vector skeleton comprises an amphiphilic copolymer with reduction sensitivity, functionalized phospholipid and cation lipid and forms a hydrophobic core, one end of the functionalized phospholipid is embedded to a hydrophilic end of the vector skeleton, the other end of the functionalized phospholipid is connected with the targeting molecules, one end of the cation lipid is embedded to the hydrophilic end of the vector skeleton, and the other end of the cation lipid is used for being connected with a therapy gene; and a Fe3O4 magnetic target, wherein the Fe3O4 magnetic target is coated in the hydrophobic core of the vector skeleton. The transported therapy gene is effectively released basedon the reduction sensitivity of the amphiphilic copolymer with reduction sensitivity; and the targeting molecules can achieve double actions of crossing blood-brain barrier and targeting glioma cells,and the targeting efficiency can be further increased by combining two kinds of targeting molecules with the Fe3O4 magnetic target.

Description

technical field [0001] The invention relates to the technical field of targeted therapy for glioma, in particular to a therapeutic gene delivery carrier and delivery system targeting glioma. Background technique [0002] Glioma is the most common central nervous system tumor, accounting for about 46% of intracranial tumors. The current treatment for glioma is mainly surgical resection, but due to its infiltrative growth, the boundary between the tumor and normal brain tissue is unclear, and it is difficult to completely remove it during surgery, which often leads to recurrence. Therefore, it is hoped that a treatment method can be developed to make up for the shortcomings that it is difficult to completely remove the lesion by surgery, and to treat glioma efficiently. Nowadays, gene therapy is considered as an effective treatment method, which intervenes in the occurrence, development and process of diseases by manipulating genetic material, including replacing or correctin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/62A61K47/52A61K47/54A61K47/60A61K47/64A61K47/69A61P35/00A61K49/06
CPCA61K48/0041A61K48/005A61K47/62A61K47/52A61K47/544A61K47/60A61K47/54A61K47/645A61K47/642A61K47/6909A61P35/00A61K49/06
Inventor 金旭乔岚歆梁艺王雅欣
Owner BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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