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Preparation method of PCADK composite microspheres encapsulating apoptotic genes

A technology of composite microspheres and genes, applied in the field of preparation of PCADK composite microspheres, can solve the problems of low gene transfection efficiency, difficulty in effectively overcoming various barriers of gene delivery, etc., so as to improve gene transfection efficiency and achieve controllable The effect of delivering, increasing the inhibition rate

Active Publication Date: 2020-01-24
SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with viral vectors, non-viral vectors are easy to be functionalized, but it is difficult to effectively overcome various barriers in the gene delivery process, and the gene transfection efficiency is low

Method used

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  • Preparation method of PCADK composite microspheres encapsulating apoptotic genes
  • Preparation method of PCADK composite microspheres encapsulating apoptotic genes
  • Preparation method of PCADK composite microspheres encapsulating apoptotic genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Preparation of BBC3@PCADK Composite Microspheres Encapsulating Apoptosis Gene

[0041] (1) Weigh 200 mg of PCADK and dissolve it in 2 mL of dichloromethane to obtain a PCADK dichloromethane solution, which is used as the oil phase for use; 20 μL of the plasmid solution containing the gene BBC3 is extracted as the inner water phase for use, wherein the plasmid concentration is 5 μg / μL Measure 60 mL, with a concentration of 1% PVA aqueous solution as the external water phase for subsequent use;

[0042] (2) Under the homogeneous condition of 13500 rpm / min, slowly drop 20 µL of the plasmid solution into the ice bath of the PCADK dichloromethane solution, and continue to stir for 2 min to obtain an emulsion;

[0043](3) The 1% PVA aqueous solution was continuously stirred at a speed of 1500 rpm / min, and then the emulsion obtained in step (2) was dropped into the above PVA aqueous solution, and the stirring was continued, and the dichloromethane was removed by volatilization...

Embodiment 2

[0046] Preparation of Bax@PCADK Composite Microspheres Encapsulating Apoptosis Gene

[0047] (1) Weigh 250 mg of PCADK and dissolve it in 3 mL of dichloromethane to obtain a PCADK dichloromethane solution, which is used as the oil phase for use; 25 μL of the plasmid solution containing the gene Bax is extracted as the inner aqueous phase for use, wherein the plasmid concentration is 5 μg / μL Measure 80 mL, with a concentration of 1% PVA aqueous solution as the external water phase for subsequent use;

[0048] (2) Under the homogeneous condition of 14500 rpm / min, slowly drop 25 µL of the plasmid solution into the ice bath of the PCADK dichloromethane solution, and continue to stir for 3 min to obtain an emulsion;

[0049] (3) The 1% PVA aqueous solution was continuously stirred at a rate of 2500 rpm / min, then the emulsion obtained in step (2) was dropped into the above-mentioned PVA aqueous solution, the stirring was continued, the dichloromethane was removed by volatilization, ...

Embodiment 3

[0051] Preparation of Noxa@PCADK Composite Microspheres Encapsulating Apoptosis Gene

[0052] (1) Weigh 100 mg of PCADK and dissolve it in 2 mL of dichloromethane to obtain a PCADK dichloromethane solution, which is used as the oil phase for use; 20 μL of the plasmid solution containing the gene Noxa is extracted as the inner water phase for use, wherein the plasmid concentration is 5 μg / μL Measure 60 mL, with a concentration of 1% PVA aqueous solution as the external water phase for subsequent use;

[0053] (2) Under the homogeneous condition of 13500 rpm / min, slowly drop 20 µL of the plasmid solution into the ice bath of the PCADK dichloromethane solution, and continue to stir for 2 min to obtain an emulsion;

[0054] (3) The 1% PVA aqueous solution was continuously stirred at a speed of 3000 rpm / min, and then the emulsion obtained in step (2) was dropped into the above PVA aqueous solution, and the stirring was continued, and the dichloromethane was removed by volatilizatio...

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Abstract

The present invention belongs to a preparation method of PCADK composite microspheres encapsulating apoptotic genes. The preparation method comprises the following specific steps: (1) weighing 200 mgof PCADK in 2 mL of dichloromethane to obtain a PCADK dichloromethane solution which is used as an oil phase for a standby application; extracting 20 [mu]L of a plasmid solution containing apoptotic genes as an internal aqueous phase for a standby application, wherein plasmid concentration is 5 [mu]g / [mu]L; and measuring 60 mL of a PVA aqueous solution at a concentration of 1% as an external aqueous phase for a standby application; (2) slowing dropping 20 [mu]L of the plasmid solution into ice bath of the PCADK dichloromethane solution at a homogenizing condition of 13,500 rpm / min and continuing stirring for 2 min to obtain an emulsion; (3) continuously stirring the 1% PVA aqueous solution at 1,500 rpm / min, then dropping the obtained emulsion in the step (2) into the above PVA aqueous solution, continuing stirring, conducting evaporation to remove the dichloromethane, then conducting centrifugation, washing, vacuum freeze-drying and storing at -20 DEG C to obtain the PCADK composite microspheres encapsulating the apoptotic genes. The PCADK is used to encapsulate the plasmid having a therapeutic effect, realizes controlled delivery of the genes and improves gene transfection efficiency.

Description

technical field [0001] The invention belongs to the technical field of medicinal materials, and in particular relates to a preparation method of PCADK composite microspheres encapsulating apoptotic genes. Background technique [0002] With the advancement of science and technology, gene therapy, a method of treating diseases at the molecular level, has attracted more and more attention of researchers. The principle is to introduce nucleic acid molecules (DNA or siRNA, etc.) Specific selectivity, regulating gene expression, and then exerting a therapeutic effect. The key to gene therapy is that genes must be effectively targeted to target cells to function efficiently. However, there are many defects in free nucleic acid molecules that limit its wide application, including poor in vivo stability, easy to be quickly cleared or degraded by nucleases, and its negative charge. The interaction with the cell membrane is hindered, and the target cells cannot be effectively reached,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63
CPCC12N15/63
Inventor 刘义王巧鲁越王云雷宇黄维孟渂偲
Owner SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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