SiRNA directed at HBx and liposome preparation thereof

A liposome and plasmid technology, which can be used in liposome delivery, DNA/RNA fragments, recombinant DNA technology, etc., and can solve problems such as low toxicity

Active Publication Date: 2014-09-10
CSPC ZHONGQI PHARM TECH (SHIJIAZHUANG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The inventors of the present invention aim at the main problems restricting the clinical application of siRNA at present, and through in-depth research, the siRNA and liposome preparations thereof loaded with HBx not only have good in vivo and in vitro stability, but also can reach target part, and obtained a higher transfection efficiency, which is conducive to its role in delivering siRNA to cells to inhibit the expression of the hepatitis B virus gene HBx. In addition, it also has lower toxicity, which is conducive to its application in vivo research

Method used

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  • SiRNA directed at HBx and liposome preparation thereof
  • SiRNA directed at HBx and liposome preparation thereof
  • SiRNA directed at HBx and liposome preparation thereof

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Embodiment 1

[0072] .

[0073] The preparation method is as follows:

[0074] 1) Blank liposome preparation: Weigh the phospholipid mixture according to the above prescription, add absolute ethanol, heat at 60°C until completely dissolved, mix with the corresponding buffer solution (volume ratio 7:13), hydrate at 60°C for 30 minutes, and complete Particles were obtained to obtain blank liposomes (containing 35% alcohol) with suitable particle size for later use.

[0075] 2) Preparation of siRNA plasmid solution: Take the concentrated solution of siRNA plasmid and dilute it with 100mM sodium acetate buffer (pH5.2) and absolute ethanol to form an siRNA solution of corresponding concentration containing 35% alcohol.

[0076] 3) Drug loading: the blank liposome and siRNA plasmid solution were mixed in equal volumes in a cross-flow manner to prepare siRNA plasmid liposomes, and incubated at 37°C for 30 minutes.

[0077] 4) Dialysis: use sucrose as the dialysate to dialyze the above-mention...

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Abstract

The invention relates to an siRNA (small interfering RNA) directed at HBx and a liposome for transferring the siRNA. The liposome contains siRNA plasmids, cationic lipid 98N12-5, cholesterol and PEGylated phospholipid. The liposome not only has good in vitro-in vivo stability, can well reach the target part, but also acquires high transfection efficiency, thus being in favor of playing a role in conveying siRNA into cells to inhibit hepatitis B virus gene HBx expression. In addition, the liposome also has low toxicity, thus being beneficial to its application in-vivo study.

Description

technical field [0001] The invention relates to the field of pharmaceutical preparations, in particular to a siRNA for HBx and a liposome for transferring the siRNA. Background technique [0002] RNA interference (RNA interfering, RNAi) refers to the phenomenon that double-stranded RNA induces specific degradation of target gene mRNA after entering cells, leading to the silencing of target gene mRNA. It is a defense system against foreign substances such as viruses formed during the long-term evolution of organisms. . siRNA (Small interfering RNA, small interfering nucleic acid) with a length of 21-23nt can trigger RNAi, can specifically knock out or shut down the expression of specific genes, and has the characteristics of high efficiency, easy synthesis and easy operation, so this technology has been Widely used in the field of gene therapy for exploring gene function and infectious diseases and malignancies. [0003] HBV (hepatitis B virus) can cause acute and chronic h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N15/66A61K48/00A61K9/127A61K47/34A61K47/18A61P31/20
Inventor 李春雷王雅娟高玉清王彩霞倪蓓蓓王艳玲
Owner CSPC ZHONGQI PHARM TECH (SHIJIAZHUANG) CO LTD
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