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Preparing process of nano c-FLIP antisense oligonucleotide particle

A technology of antisense oligonucleotides and nanoparticles, which can be used in medical preparations of non-active ingredients, pharmaceutical formulas, gene therapy, etc., and can solve the problems of large average particle size, polyvinyl alcohol, and low drug loading , to achieve the effect of small average particle size, volatile and stable physical properties

Inactive Publication Date: 2006-11-08
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
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  • Claims
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Problems solved by technology

Journal of Second Military Medical University, 2005, 26 (5): 547-550.] This method is practical and effective, simple and easy, but there are many situations of organic solvent and polyvinyl alcohol (PVA) remaining in the preparation process, they are harmful to Normal cells have certain toxicity; at the same time, the prepared nanoparticles have the defects of wide particle size distribution (50-150nm), large average particle size (95.5nm), and low drug loading (0.579%)

Method used

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Examples

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Effect test

Embodiment 1

[0020] Example 1: Preparation of c-FLIP antisense oligonucleotide nanoparticles by double emulsion-drying method

[0021] Take 660 μg of antisense oligonucleotides dissolved in 150ul of buffer solution (PBS pH=7.4) as the inner water phase, dissolve 50mg of PLGA (RG503H) in 1.5ml of dichloromethane as the oil phase, and add the inner water phase to Ultrasonic emulsification of the oil phase in an ice bath for 20s (power 20W) to obtain colostrum, add 3ml of 1% (w / v) poloxamer188 aqueous solution to the colostrum, continue ultrasonic emulsification in an ice bath for 30s (power 20W) to form double emulsion, and double emulsion Pour into 30ml of 0.3% (w / v) PVA aqueous solution, stir at a low speed at room temperature for 4 hours (stirring speed is 400rpm), volatilize and remove the organic solvent, collect nanoparticles by centrifugation, wash with ultrapure water three times, and freeze-dry to obtain the present invention. nanoparticles.

[0022] The nano-particles obtained by ...

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Abstract

The present invention relates to medicine technology, and especially preparation process of nanometer c-FLIP antisense oligonucleotide particle. The present invention prepares nanometer c-FLIP antisense oligonucleotide particle preparation through a composite emulsifying-drying process, which includes dissolving proper amount of c-FLIP-ASODN in super pure water to form water phase, dissolving proper amount of lactic acid-hydroxyaceteic acid copolymer in dichloromethane to obtain oil phase, mixing the water phase and the oil phase and ultrasolic emulsifying in ice bath to obtain initial emulsion, adding Poloxamer-188 solution and re-emulsifying to obtain composite emulsion, adding the composite emulsion to PVA solution, volatilizing organic solvent at room temperature through stirring, centrifuging precipitating, water washing, and freeze drying to obtain the powder.

Description

technical field [0001] The invention relates to the technical field of medicine, and relates to a preparation method of c-FLIP antisense oligonucleotide nanoparticles. Background technique [0002] Antisense oligonucleotides (antisense oligodeoxynucleotides, ASODN) are artificially synthesized DNA molecules and their analogs with a length of 10-30 bases, which specifically bind to specific mRNA or DNA through Watson-Crick base pairing , so as to inhibit or block the expression of the gene, make it lose its activity, and achieve the purpose of gene therapy. c-FLIP protein is an inhibitory protein in the process of cell apoptosis. It has been confirmed that c-FLIP is expressed in a variety of tumors, thereby causing tumor cells to have apoptosis disorders. c-FLIP-ASODN sequence is: 5'- ACT TG T CCC TGC TCC TTG AA -3′, Mr=6 000, the underlined part is thio modification to improve its stability [Bannerman DD, Tupper JC, Ricketts WA, et al.Aconstitut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/14A61K9/19A61K47/10A61K47/34A61K48/00A61P35/00
Inventor 魏锐利聂有华马晓晔朱煌梁莉
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY