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Respiratory tract homing siRNA atomizing nanometer drug delivery system and preparation method thereof

A technology of drug delivery system and respiratory tract, applied in respiratory diseases, pharmaceutical formulations, drug combinations, etc., can solve problems such as structural damage of nucleic acid substances, and achieve the effect of improving the efficiency of uptake and gene transfection

Inactive Publication Date: 2011-09-14
广州医学院 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the problem is that high-frequency oscillation will damage the structure of nucleic acid substances, so it is difficult to adopt the inhalation treatment mode for small molecule RNA interference treatment of respiratory diseases in clinic

Method used

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  • Respiratory tract homing siRNA atomizing nanometer drug delivery system and preparation method thereof
  • Respiratory tract homing siRNA atomizing nanometer drug delivery system and preparation method thereof
  • Respiratory tract homing siRNA atomizing nanometer drug delivery system and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0090] Experimental example 1: the ability of Sal-guanidinylated chitosan nanoparticles to transport nucleic acids in vivo

[0091] main experimental materials

[0092] Natural chitosan nanoparticles: American Sigma company

[0093] Sal-guanidinated chitosan: prepared according to the method described in "Salbutamol modified guanidinated chitosan and its preparation method and application" (application number: 200910245193.6)

[0094] MEM medium: American Gibico company

[0095] Fetal bovine serum: Hangzhou Sijiqing Company

[0096] Plasmid extraction kit: Promega, USA

[0097] Lipofectamin 2000 (nucleic acid transfection reagent): American Invitrogen Company

[0098] siRNA: American AMBION company

[0099] Yoyo1 (fluorescent dye, used to label nucleic acid substances): Invitrogen, USA

[0100] experimental method

[0101] 1. Sal-guanidinylated chitosan nanoparticles transport nucleic acids into cells

[0102] 1.1 Take 1 ug of siRNA and Yoyo1 fluorescent dye to complex...

experiment example 2

[0110] Experimental example 2: Sal-guanidinylated chitosan and siRNA composite nanoparticles exert RNA interference effect in vivo

[0111] main experimental materials

[0112] Sal-guanidinated chitosan: prepared according to the method described in "Salbutamol modified guanidinated chitosan and its preparation method and application" (application number: 200910245193.6)

[0113] Natural chitosan nanoparticles: American Sigma company

[0114] Green fluorescent protein (GFP) siRNA: American AMBION company

[0115] Negative Control siRNA: American AMBION company

[0116] HEK293 cell line stably expressing GFP: screening and cultivation in our laboratory

[0117] EGFP transgenic mice: Saiye Biotechnology Co., Ltd. The EGFP transgenic mice are in the C57BL / 6 background, and the expression method is systemic expression. The EGFP gene was constructed on the pCAGGS vector, which has chicken β-actin promoter and CMV enhancer.

[0118] Ultrasonic atomizing head (Aeroneb Lab Neb...

experiment example 3

[0129] Experimental example 3: The ability of the composite nanoparticle of Sal-guanidinylated chitosan and nucleic acid to target and transport nucleic acid for RNA interference in vivo

[0130] Materials and methods

[0131] main experimental materials

[0132] Guanidinated chitosan: prepared according to the method described in "Salbutamol Modified Guanidinated Chitosan, Preparation Method and Application" (Chinese patent application, application number: 200910245193.6).

[0133] Salbutamol (Sal): American Sigma Company.

[0134] Sal-guanidinated chitosan nanocarrier: prepared according to the method described in "Salbuterol Modified Guanidinated Chitosan and Its Preparation Method and Application" (Chinese patent application, application number: 200910245193.6).

[0135] 16HBE cells (human bronchial epithelial cell line): a gift from Professor Steven.Holgate, University of Southampton, UK.

[0136] pEGFP-N2 plasmid: American Clontech Company.

[0137] Green Fluorescent...

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Abstract

The invention discloses a respiratory tract homing siRNA atomizing nanometer drug delivery system which uses salbutamol guanidinated chitosan treated by ultrasonic atomization as a carrier material, and is prepared by wrapping siRNA which interferes with target pathogenic genes. The invention also discloses a preparation method of the respiratory tract homing siRNA atomizing nanometer drug delivery system. The drug delivery system disclosed by the invention can be used to deliver siRNA which interferes with target pathogenic genes so as to play an RNA interference role and inhibit the replication of the pathogenic genes, can deliver siRNA with the target of lung respiratory tract epithelia in vivo, and is a non-virus delivery system which exerts corresponding drug effect by aiming at silence the target genes.

Description

technical field [0001] The invention relates to a drug delivery system, in particular to a respiratory tract homing siRNA atomized nano drug delivery system. The present invention also relates to the preparation method of the above respiratory tract homing siRNA atomized nano drug delivery system. Background technique [0002] RNA interference (interfering RNAs, RNAi) is one of the most promising areas of gene therapy research. As far as the general law of biology is concerned, RNAi can silence any gene. At the same time, due to the high specificity, efficiency and simplicity of RNAi-mediated gene silencing, it has great potential in the treatment of diseases caused by pathogen invasion such as virus infection or gene mutation. Obvious clinical application prospects. Studies have shown that the use of siRNA of certain structural protein genes of SARS coronavirus can inhibit virus replication in the lungs of macaques infected with SARS coronavirus. In addition, the use of ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K9/12A61K47/48A61P11/00A61K47/61
Inventor 徐军罗永峰刘文广翟欣昀孙鹏
Owner 广州医学院
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