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Gene medicine conveying system and method of preparing the same

A gene drug and delivery system technology, applied in drug delivery, gene therapy, powder delivery, etc., can solve the problems of the stability of gene drugs and the inevitable degradation of gene drugs, so as to enhance targeting and tissue specificity, prolong Plasma half-life, cytotoxicity reduction effect

Inactive Publication Date: 2008-03-12
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the factors involved in the preparation process of the above-mentioned nanoparticles include emulsification energy, organic solvent, surface tension between organic phase / water phase, molecular weight and concentration of polymers, etc. will all have an impact on the stability of gene medicines.
In order to control the influence of the preparation process, technologies such as spray drying and reverse solvent diffusion are widely used, but the degradation of gene drugs is still inevitable

Method used

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  • Gene medicine conveying system and method of preparing the same
  • Gene medicine conveying system and method of preparing the same

Examples

Experimental program
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Effect test

preparation Embodiment 1

[0052] 120 μg of DNA was dissolved in 500 μl of deionized water, and added to 100 μl of an aqueous solution containing 72 μg of poly-L-lysine and 30 μg of chloroquine under vortex mixing, mixed well, and allowed to stand at room temperature for 30 minutes.

[0053] 100mg of PEG-polycyanoacrylate copolymer was dissolved in 2ml of dichloromethane to form an organic phase, mixed with the above complex solution, ultrasonicated for 5 seconds in an ice bath, and the formed W / O colostrum was dispersed into 10ml of 1.0% PVA In the aqueous solution, ultrasonic 5 seconds in the ice bath, the W / O / W type double emulsion that forms continues to disperse in the aqueous solution of 20ml 0.3%PVA, and magnetic force stirs, and organic solvent is evaporated, and the nanoparticle that forms is solidified. Nanoparticles were collected by centrifugation at 39,000×g for 20 minutes.

[0054] 80 mg of transferrin was dissolved in 1 ml of 30 mM sodium acetate buffer (pH 5), and 50 μl of sodium acetate...

preparation Embodiment 2

[0057]50 μg of DNA was dissolved in 300 μl of deionized water, and added to 50 μl of an aqueous solution containing 5 μg of polyethyleneimine under vortex mixing, thoroughly mixed, and allowed to stand at room temperature for 30 minutes.

[0058] Dissolve 80mg of PLGA-PEG in 2ml of chloroform to form an organic phase, mix with the above complex solution, and ultrasonicate for 5 seconds in an ice bath. Seconds, the formed W / O / W type double emulsion continued to be dispersed into 20ml of 0.2% HS15 aqueous solution, stirred by magnetic force, and the organic solvent was evaporated to solidify the formed nanoparticles. Nanoparticles were collected by centrifugation at 39,000×g for 20 minutes.

[0059] 40mg of folic acid was dissolved in 2ml of DMSO, 30mg of NHS, 20mg of DCC and a few drops of triethylamine were added, protected from light, and incubated for 90 minutes.

[0060] 0.4 μmol of NHS-folate was quickly mixed with 0.4 ml of PBS buffer containing 10 mg of the above-mentio...

preparation Embodiment 3

[0062] 20 μg of DNA was dissolved in 200 μl of deionized water, and added to 80 μl of an aqueous solution containing 60 μg of protamine sulfate and 30 μg of chloroquine under vortex mixing, thoroughly mixed, and allowed to stand at room temperature for 30 minutes.

[0063] Dissolve 30mg of phosphatidylethanolamine in 1ml of ethanol to form an organic phase, mix with the above complex solution, and ultrasonicate for 5 seconds in an ice bath, and the formed W / O colostrum is dispersed into 10ml of 2.0% aqueous solution of sodium fatty acid sulfonate, and placed in an ice bath Ultrasonic for 5 seconds, the formed W / O / W type double emulsion continued to be dispersed in 20ml of 0.5% aqueous solution of sodium fatty acid sulfonate, magnetically stirred, and the organic solvent was evaporated to solidify the formed nanoparticles. Nanoparticles were collected by centrifugation at 39,000×g for 20 minutes.

[0064] 0.2 μmol of activated RGD peptide was quickly mixed with 0.5 ml of PBS bu...

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Abstract

The present invention relates to a transportation system for the genetic medicine and the preparation method. The transportation system is a composite consisting of the genetic medicine, the cationic peptide or the polymer and any selected adjuvant ingredient. The composite is packaged in a nano particle of polymer material modified by the PEG. The ligand modification is conducted in the nano particle surface. The preparation method comprises the following steps. Step one, the genetic medicine, the cationic peptide or the polymer and any selected adjuvant ingredient are mixed in order to prepare the composite solution; step two, the organic solution of the polymer material is added into the composite solution and dispersed into the water phase, therefore the W / O / W-typed multiple emulsion is prepared, followed by separation of the collected nano particles; step three, the nano particle surface modified by the ligand and the collected nano particles are separated after modification. The stability of the genetic medicine in the preparation process and the encapsulation efficiency are both improved in the system. The surface modification not only prolongs the plasma half-life and improves the targeting property in the transportation system, but also enhances the transfection efficiency and greatly reduces the cytotoxicity in the transportation system, which therefore guarantees the safety of the medicine application.

Description

technical field [0001] The invention relates to the field of pharmaceutical preparations. Specifically, the present invention relates to a new gene drug delivery system and its preparation method. Background technique [0002] Nucleic acid substances such as plasmid DNA, antisense RNA, and oligodeoxynucleotides, as gene therapy drugs and vaccines, have shown great advantages and potentials in the treatment of major diseases such as tumors and hemophilia and genetic diseases. [0003] Naked DNA is easily degraded by nucleases, and after intravenous injection into the body, it is quickly cleared from the plasma, so it needs an appropriate carrier to protect and transport it. Due to safety hazards such as immunogenicity and potential pathogenicity, viral vectors are gradually being replaced by non-viral vectors. [0004] Traditional non-viral vectors such as liposomes, nanoparticles, etc., after entering the body, due to the interaction with plasma proteins, they are easily s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K9/14A61K9/00
Inventor 李亚平顾王文陈伶俐
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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