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Targeted gene carrier material based on graphene oxide and its preparation and application

A gene carrier and graphene technology, applied in gene therapy, medical preparations of non-active ingredients, inorganic non-effective ingredients, etc., can solve the problems of high toxicity of siRNA carriers, no clinical application value, low transfection efficiency, etc. Achieve effects that are not easy to be degraded, easy to modify, and conducive to release

Inactive Publication Date: 2011-12-14
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a targeted gene carrier material based on graphene oxide and its preparation and application. It is aimed at the existing siRNA carrier with high toxicity, low transfection efficiency and instability, and no clinical application value. Insufficiency of using functionalized graphene oxide as a targeted delivery carrier for siRNA and other gene molecules, providing a low-toxicity and high-efficiency siRNA transfection agent, which can be delivered into tumor cells, so as to achieve the purpose of inhibiting the expression of target genes and related proteins

Method used

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  • Targeted gene carrier material based on graphene oxide and its preparation and application
  • Targeted gene carrier material based on graphene oxide and its preparation and application
  • Targeted gene carrier material based on graphene oxide and its preparation and application

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Embodiment 1

[0035] The first step: synthesis of graphene oxide materials, the preparation process can refer to (ACSNano, 2008, 2, 463-470); the core of this step is to obtain graphene oxide materials. Similarly, other methods to obtain graphene oxide materials can also be used.

[0036] The second step: preparation of polyethylene glycol and folic acid conjugate:

[0037] Mix 44mg of folic acid and 420mg of amino-terminated polyethylene glycol (Mn=2100) in a mixed solvent of 40mL of N,N-dimethylsulfoxide and 10mL of water, add 50mg of 1-ethyl-3 (3-dimethylamino -propyl)carbodiimide (EDC), 30mg N-hydroxy succinimide (NHS) was stirred at room temperature for one day, the system was rotary evaporated to remove the solvent, the concentrate was passed through a Sephadex column, the yellow solution that flowed out first was collected, and the product was concentrated and freeze-dried to obtain The coupling product of the two.

[0038] The third step: graphene oxide is connected with the produ...

Embodiment 2

[0051] The first step: synthesis of graphene oxide materials, the preparation process can refer to (ACSNano, 2008, 2, 463-470); the core of this step is to obtain graphene oxide materials. Similarly, other methods to obtain graphene oxide materials can also be used.

[0052] The second step: preparation of polyethylene glycol and folic acid conjugate:

[0053] Mix 44mg of folic acid and 300mg of amino-terminated polyethylene glycol (Mn=1500) in a mixed solvent of 40mL N,N-dimethylsulfoxide and 10mL of water, add 50mg of 1-ethyl-3 (3-dimethylamino -propyl)carbodiimide (EDC), 30mg N-hydroxy succinimide (NHS) was stirred at room temperature for one day, the system was rotary evaporated to remove the solvent, the concentrate was passed through a Sephadex column, the yellow solution that flowed out first was collected, and the product was concentrated and freeze-dried to obtain The coupling product of the two.

[0054] The third step: graphene oxide is connected with the product ...

Embodiment 3

[0063] The first step: synthesis of graphene oxide materials, the preparation process can refer to (ACSNano, 2008, 2, 463-470); the core of this step is to obtain graphene oxide materials. Similarly, other methods to obtain graphene oxide materials can also be used.

[0064] The second step: preparation of polyethylene glycol and folic acid conjugate:

[0065] Mix 44mg of folic acid and 420mg of amino-terminated polyethylene glycol (Mn=2100) in a mixed solvent of 40mL of N,N-dimethylsulfoxide and 10mL of water, add 50mg of 1-ethyl-3 (3-dimethylamino -propyl)carbodiimide (EDC), 30mg N-hydroxy succinimide (NHS) was stirred at room temperature for one day, the system was rotary evaporated to remove the solvent, the concentrate was passed through a Sephadex column, the yellow solution that flowed out first was collected, and the product was concentrated and freeze-dried to obtain The coupling product of the two.

[0066] The third step: graphene oxide is connected with the produ...

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Abstract

The invention relates to a tumor-targeted gene vector material and a preparation method and use thereof. In the invention, functional graphene oxide is used as a targeted transport vector of genomic molecules such as siRNA. The preparation method comprises: connecting folic acid tumor cell targeted molecules and amino-terminated polyethylene glycol by amido bonds; connecting the graphene oxide with the coupling material by amido bonds to obtain the tumor targeted functional graphene oxide; anchoring aminated molecules with an aromatic conjugated structure onto the surface of the graphene oxide through pi-pi conjugation; and finally, preparing the tumor targeted high-efficiency gene transport system by static attraction between the positively charged functional graphene oxide and the negatively charged genomic molecules. The vector material has a high targeting performance for tumor cells in which folic acid receptor is expressed more highly, and after being loaded with siRNA, the corresponding target gene level and related protein expression can be reduced. The invention provides a new method for the in-vivo targeted transportation of genes.

Description

technical field [0001] The present invention relates to a targeting gene carrier material and its preparation and application, in particular to a novel tumor-targeting small interfering RNA carrier material based on functionalized graphene oxide and its preparation method and application. Background technique [0002] RNA interference therapy is currently the most potential and effective genetic therapy. It is a gene silencing mechanism in which double-stranded RNA specifically inhibits the expression of its complementary gene, and is a homologous mRNA degradation phenomenon mediated by double-stranded RNA in various organisms. The mechanism of small interfering ribonucleic acid (small interfering dsRNA, siRNA) molecules blocking gene expression is because it targets mRNAs that are homologous and complementary to the sequence, and degrades specific mRNAs, resulting in gene silencing effects. However, siRNA itself is unstable, easily degraded, and difficult to penetrate into...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K48/00A61P35/00A61K47/34A61K47/22A61K47/02A61K47/26A61K47/18
Inventor 杨晓英段宏泉王银松陈永胜马延风黄毅
Owner TIANJIN MEDICAL UNIV
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