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Novel albumin-siRNA composite nanometer particle, and preparation method and applications thereof

A composite nanoparticle and albumin technology, which is applied in the field of novel albumin-siRNA composite nanoparticles and its preparation, can solve the problems that have not yet been reported on the oral drug delivery targeting system of albumin and siRNA composite nanoparticles, and achieve Maintain solubility and tumor-targeted enrichment properties, simple operation, and good dispersion

Active Publication Date: 2018-09-04
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no relevant research report on the oral drug delivery targeting system based on the preparation of albumin and siRNA composite nanoparticles by this method

Method used

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  • Novel albumin-siRNA composite nanometer particle, and preparation method and applications thereof
  • Novel albumin-siRNA composite nanometer particle, and preparation method and applications thereof
  • Novel albumin-siRNA composite nanometer particle, and preparation method and applications thereof

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Embodiment 1

[0037]The preparation of embodiment 1 albumin-siRNA nano drug delivery system (HSA-siRNA NP):

[0038] (1) Stir 200mg / mL albumin solution and 20mM glutathione solution (phosphate buffered saline solution of glutathione, pH 5.0-10.0) at 20°C for 100min to obtain the final protein A homogeneous solution of albumin with a spatial structure expansion at a concentration of 150 mg / mL;

[0039] (2) Add 5 mM siRNA (targeting the GRP78 gene as an example, the same below) DEPC aqueous solution and 20 mM sodium selenite solution to the albumin homogeneous solution in which the spatial structure is unfolded to obtain albumin in the albumin solution The final concentration was 120mg / mL, and stirred at 4°C for 6h to obtain a crude solution of albumin-siRNA nanoparticles;

[0040] (3) Put the crude solution into a dialysis bag, dialyze molecular cut-off not less than 1000, and dialyze in PBS solution at 0-20°C to remove excess glutathione, selenium compounds and their by-products to obtain ...

Embodiment 2

[0042] Example 2 Preparation of albumin-SH-siRNA nano drug delivery system (HSA-(SH) siRNA NP):

[0043] (1) Mix 180mg / mL albumin solution and 50mM glutathione solution (phosphate buffer solution of glutathione, pH value 5.0-10.0) at 30°C for 60min to obtain the final protein A homogeneous solution of albumin with a spatial structure expansion at a concentration of 130 mg / mL;

[0044] (2) Add 6mM DEPC aqueous solution of SH-siRNA and 30mM sodium selenite solution to the albumin homogeneous solution with spatial structure unfolding to obtain a final albumin concentration of 100mg / mL in the albumin solution, stir at 4°C 16h, to obtain a crude solution of albumin-siRNA nanoparticles;

[0045] (3) Put the crude solution into a dialysis bag, dialyze molecular cut-off not less than 1000, and dialyze in PBS solution at 0-20°C to remove excess glutathione, selenium compounds and their by-products to obtain albumin-(SH ) siRNA composite nanoparticles (Note: During the preparation pro...

Embodiment 3

[0047] Example 3 Preparation of albumin-SH-siRNA nano drug delivery system (HSA-(SH) siRNA NP):

[0048] (1) Stir 80mg / mL albumin solution and 30mM glutathione solution (phosphate buffered saline solution of glutathione, pH 5.0-10.0) at 34°C for 50min to obtain the final protein A homogeneous solution of albumin with a spatial structure expansion at a concentration of 60 mg / mL;

[0049] (2) Add 4mM DEPC aqueous solution of SH-siRNA and 10mM sodium selenite solution to the albumin homogeneous solution developed in spatial structure to obtain a final albumin concentration of 40mg / mL in the albumin solution, stir at 4°C 24h, to obtain a crude solution of albumin-(SH)siRNA nanoparticles;

[0050] (3) Put the crude solution into a dialysis bag, dialyze molecular cut-off not less than 1000, and dialyze in PBS solution at 0-20°C to remove excess glutathione, selenium compounds and their by-products to obtain albumin-(SH ) siRNA composite nanoparticles.

[0051] Albumin-(SH)siRNA c...

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Abstract

The invention belongs to the technical field of biological medicine, and discloses a novel albumin-siRNA composite nanometer particle, and a preparation method and applications thereof. According to the preparation, the internal space structure of albumin is destroyed using a disulfide bond reducing agent, and proteins containing mercapto group active groups are obtained, siRNA and / or a modified substance SH-siRNA is added, and a selenium compound is added so as to obtain the novel albumin-siRNA composite nanometer particle. The preparation method is simple; the nanometer size is uniform at different dilution concentrations, the dispersion performance is excellent, the stability in artificial gastric juice, intestinal juice, and blood environment is excellent; and in addition, the novel albumin-siRNA composite nanometer particle possesses intracellular reduced glutathione microenvironment responsiveness; the novel albumin-siRNA composite nanometer particle possesses oral administrationand injection administration living tumor targeting characteristic, is capable of increasing siRNA anti-cancer drug oral administration or injection administration anti-cancer activity, and reducingtoxic or side effect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a novel albumin-siRNA composite nanoparticle and its preparation method and application. Background technique [0002] Albumin is a biological endogenous protein with many characteristics such as biodegradability, non-toxicity, and non-antigenicity. It is considered to be an ideal drug carrier. research direction. The combined albumin drug delivery system is a very ideal drug loading mode. Encapsulating drug molecules in albumin nanoparticles can significantly improve the stability and stability of water-insoluble drugs in aqueous solution (that is, in the blood circulation system in vivo). Solubility. At the same time, the enhanced permeability and retention effect (EPR effect) of tumor tissue can be used to make the albumin nano-drug delivery system achieve the purpose of targeted drug delivery. In addition, albumin can improve the solubility and blood circul...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K33/04A61K31/7088A61K47/64
CPCA61K31/7088A61K33/04A61K47/6931
Inventor 谭蔚泓彭咏波刘腾李雄
Owner HUNAN UNIV
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