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Method for preparing DNA nanopreparation used for treating tumors via combination of immunotherapy and chemotherapy

A technology of combination therapy and nano-formulation, applied in anti-tumor drugs, drug combinations, pharmaceutical formulations, etc., can solve problems such as limiting the application of gel particles, and achieve the effects of improving biocompatibility, avoiding potential toxicity and sensitive induction.

Inactive Publication Date: 2020-04-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Now most of the chemical reagents are used as the cross-linking agent of microgel particles, but the potential toxicity of chemical reagents limits the application of gel particles in the human body

Method used

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  • Method for preparing DNA nanopreparation used for treating tumors via combination of immunotherapy and chemotherapy

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of DNA-A@nucleic acid aptamer polymer chain

[0032] (1) At room temperature, mix 4uL 15mg / ml DNA-A aqueous solution, 3uL 5mg / ml nucleic acid aptamer aqueous solution, 2uL 1mg / mL N-isopropylacrylamide, 1uL 1% 2-carboxy-40-( 2-Carboxyethoxy)-2-methyl-propiophenone initiator aqueous solution, 15uL of Tris buffer (pH=8.0) and 25uL of ultrapure water were mixed, and stirred at a speed of 400rpm;

[0033] (2) Then the mixed solution was irradiated for 5 min under a purple light with a wavelength of 365 nm.

Embodiment 2

[0035] Preparation of DNA-B polymer chains

[0036] (1) At room temperature, mix 10uL 10mg / ml CpG-ODN aqueous solution, 1uL 1mg / mL N-isopropylacrylamide, 1uL 1% 2-carboxy-40-(2-carboxyethoxy)-2- Methyl-propiophenone initiator aqueous solution, 25uL of Tris buffer (pH=8.0) and 25uL of ultrapure water were mixed, and stirred at a speed of 400rpm;

[0037] (2) Then the mixed solution was irradiated for 5 min under a purple light with a wavelength of 365 nm.

Embodiment 3

[0039] Preparation and collection of DNA microgels

[0040] (1) At room temperature, fully mix 50uL DNA-A polymer chain solution, 5uL CpG-ODN polymer chain solution and 5u 5mg / ml ligated DNA aqueous solution to obtain a mixed solution.

[0041] (2) The mixed solution was incubated in a water bath at 80° C. for 5 min, and then slowly cooled to room temperature.

[0042] (3) Put the obtained microgel solution into a dialysis bag with a molecular weight cut-off of 8000, dialyze in Tris buffer for 2-3 days, finally collect by centrifugation, and wash with water three times.

[0043] (4) The collected microgel particles were freeze-dried in a vacuum oven at -55°C to a powder state.

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Abstract

The invention discloses a method for preparing a DNA nanopreparation used for treating tumors via a combination of immunotherapy and chemotherapy. The method mainly comprises the following steps: (1)preparing a DNA-A@nucleic acid aptamer polymer chain; (2) preparing a CPG-ODN polymer chain; (3) preparation of a DNA microgel nanoparticle through cross-linking of the DNA-A polymer chain, the CPG-ODN polymer chain and Linker DNA since the DNA-A polymer chain and the CPG-ODN polymer chain are in base complementary with the Linker DNA base, wherein the DNA-A and CPG-ODN are linked with the LinkerDNA by utilizing a DNA base complementary pairing principle so as to form the microgel particles through crosslinking; and 4) loading of a chemotherapeutic drug, i.e., DOX. A DNA-A solution is mixed with an aptamer to form the DNA-A polymer chain with the aptamer. The microgel nanoparticle combines chemotherapy and immunotherapy together, so tumor treatment effect is greatly enhanced, and tumor treatment is more thorough.

Description

technical field [0001] The invention relates to the technical field of preparation preparation, specifically a method for synthesizing nano-preparations for tumor treatment by using the principle of DNA base complementary pairing to link DNA-A and CpG-ODN with connecting DNA. Background technique [0002] Microgels are polymer colloid particles with a size of micron or nanometer, which have the characteristics of high loading efficiency, good biocompatibility and excellent biostability. Moreover, microgels are more sensitive to external stimuli, and can quickly respond to changes in the external environment through large deformations. These characteristics make microgels have great advantages and potentials in the field of drug delivery. Nowadays, chemical reagents are mostly used as cross-linking agents of microgel particles, but the potential toxicity of chemical reagents limits the application of gel particles in the human body. [0003] The invention adopts the mechani...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K47/58A61K9/06A61P35/00A61K31/704
CPCA61K31/704A61K39/39A61K47/58A61K9/06A61K9/0024A61P35/00A61K2039/55561A61K2300/00
Inventor 郑斌彭文畅明东刘爽甘霖
Owner TIANJIN UNIV
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