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siRNA (small interfering ribose nucleic acid) micro emulsion carrier and preparation method of siRNA micro emulsion carrier

A technology of microemulsion and carrier, which is applied in the field of medicine to achieve the effects of prolonging residence time, improving stability and protecting stability

Inactive Publication Date: 2012-12-05
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no literature report on an siRNA carrier that can protect the stability of siRNA and facilitate its smooth penetration through the skin or mucous membrane

Method used

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  • siRNA (small interfering ribose nucleic acid) micro emulsion carrier and preparation method of siRNA micro emulsion carrier
  • siRNA (small interfering ribose nucleic acid) micro emulsion carrier and preparation method of siRNA micro emulsion carrier
  • siRNA (small interfering ribose nucleic acid) micro emulsion carrier and preparation method of siRNA micro emulsion carrier

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Preparation of NC-FAM siRNA microemulsion

[0039] In order to prevent RNase contamination, all medicines, containers, and materials used in the microemulsion preparation process must be treated without enzymes or must be enzyme-free products. The oil phase and two emulsifiers used in the preparation of microemulsions were baked in an oven at 75°C for 8 hours; metal containers were baked in an oven at 180°C for 3 hours; plastics, rubber and other materials that are not resistant to high temperatures were soaked in 0.1% DEPC water 24h, can withstand high temperature and high pressure and then 120 ℃, 20min sterilization; disposable materials use enzyme-free products.

[0040] NC-FAM siRNA microemulsion formulation and ratio:

[0041]

[0042] The preparation method is as follows: design siRNA (NC siRNA) of any sequence, modify it with carboxyfluorescein (FAM) to form NC-FAM siRNA, and dissolve it in enzyme-free water. Weigh the oil, emulsifier and co-em...

Embodiment 2

[0044] Embodiment 2: Preparation of eotaxin siRNA microemulsion

[0045] In order to prevent RNase contamination, all medicines, containers, and materials used in the microemulsion preparation process must be treated without enzymes or must be enzyme-free products. The oil phase and two emulsifiers used in the preparation of microemulsions were baked in an oven at 75°C for 8 hours; metal containers were baked in an oven at 180°C for 3 hours; plastics, rubber and other materials that are not resistant to high temperatures were soaked in 0.1% DEPC water 24h, can withstand high temperature and high pressure and then 120 ℃, 20min sterilization; disposable materials use enzyme-free products.

[0046] Formula and ratio of Eotaxin siRNA microemulsion:

[0047]

[0048]

[0049] The preparation method is: design eotaxin-specific siRNA, i.e. eotaxin siRNA (design according to the Mus musculus small chemokine (C-C motif) ligand 11 (CCL11) mRNA (982bp) whose sequence Accession Num...

Embodiment 3

[0050] Embodiment 3: Preparation of STAT6siRNA microemulsion

[0051] SATA6siRNA microemulsion formula and ratio:

[0052]

[0053]STAT6 (Signal transducers and activators of transcription 6, signal transduction and transcription activator 6)-specific siRNA, that is, STAT6siRNA, the sequence refers to the report of Rippmann JF et al. (Rippmann JF, Schnapp A, Weith A, Hobbie S. Gene silencing with STAT6specific siRNAs blocks eotaxin release in IL-4 / TNFalpha stimulated human epithelial cells. FEBS Lett.2005Jan 3; 579(1): 173-8.), Sense strand: 5'CAG UUC CGC CAC UUG CCA ATT 3'; Antisense strand: 5'UUG GCA AGU GGC GGA ACU GTT3'. All the other are with embodiment 2.

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Abstract

The invention relates to the technical field of medicine. No siRNA (small interfering ribose nucleic acid) percutaneous administration mode exists in the prior art for lack of a percutaneous administration transfer system. The invention aims at providing a percutaneous administration transfer system of a siRNAmicro emulsion carrier capable of encapsulating the siRNA of relevant genes of specific diseases into the siRNA micro emulsion carrier. The siRNAmicro emulsion carrier protects the stability of the siRNA encapsulated inside the siRNAmicro emulsion carrier, and in addition, the percutaneous capability is high. The invention has another goal of providing a preparation method of the siRNA micro emulsion carrier. The invention provides the siRNA micro emulsion carrier with the proportion of oil phase: emulsifying agents: co-emulsifying agents: water phase being (40-70 percent):(10-20 percent):(10 to 20 percent):(5 to 20 percent). The siRNA micro emulsion carrier has the advantages that macromolecular siRNA can be encapsulated into the water phase, the siRNA is brought into the skin and the mucosa through the skin and the mucosa, the siRNA percutaneous transfer problem is solved, meanwhile, the stability of the siRNA is greatly improved, and the siRNA micro emulsion carrier and the preparation method can be used for siRNA medicine development and scientific research application.

Description

technical field [0001] The invention relates to the field of medical technology, and relates to a transdermal delivery system in which siRNA of a specific disease-related gene can be encapsulated—siRNA microemulsion carrier and a preparation method thereof. Background technique [0002] Gene therapy is a major breakthrough in the concept, technology and method of disease treatment. The discovery of RNA interference and the application of siRNA provide a new method for gene therapy and enrich the content of gene drug development. The phenomenon of RNA interference (RNA interfering, RNAi) is that double-stranded RNA (double-stranded RNA, dsRNA) homologous to the target gene sequence is cleaved into small interfering RNA (small interfering RNA, siRNA) consisting of 21-25 nucleotides. ) and the post-transcriptional silencing process of sequence-specific genes widely present in organisms (see literature ① Hannon GJ. RNA interferences. Nature. 2002 Jul 11; 418(6894): 244-51. . Ki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/107A61K48/00A61K47/34A61K47/14
Inventor 胡晋红彭程
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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