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Method for preparing DNA nano preparation for photo-thermal-immune combined treatment of tumors

A technology of combination therapy and nano-formulation, which can be used 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 injury, and reducing volume.

Inactive Publication Date: 2020-04-17
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 nano preparation for photo-thermal-immune combined treatment of tumors

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Preparation of DNA-A@nucleic acid aptamer polymer chain

[0033] (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;

[0034] (2) Then the mixed solution was irradiated for 5 min under a purple light with a wavelength of 365 nm.

Embodiment 2

[0036] Preparation of CpG-ODN polymer chains

[0037] (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;

[0038] (2) Then the mixed solution was irradiated for 5 min under a purple light with a wavelength of 365 nm.

Embodiment 3

[0040] Preparation and collection of DNA microgels

[0041] (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.

[0042] (2) The mixed solution was incubated in a water bath at 80° C. for 5 min, and then slowly cooled to room temperature.

[0043] (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.

[0044] (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 nano preparation for photo-thermal-immune combined treatment of tumors. The method mainly comprises the following steps: (1) a DNA-A and nucleic acid aptamer polymer chain is prepared; (2) a CPG-ODN polymer chain is prepared; (3) the DNA-A polymer chain and the CPG-ODN polymer chain are complementary with a Linker DNA base, and the chains and the Linker DNA base can be crosslinked to form DNA microgel nanoparticles, and meanwhile, an indocyanine green (ICG) photo-thermal agent is added, and ICG-coated microgel particles are formed through crosslinking; and (4) loading of a chemotherapeutic drug DOX is carried out. A nucleic acid aptamer is mixed in a DNA-A solution to form a DNA-A polymer chain with a nucleic acid aptamer. The microgel nanoparticles combine chemotherapy and immunotherapy, so that the 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 of nanometer preparations, in particular to a method for synthesizing nanometer preparations for tumor treatment by using the principle of DNA base complementary pairing to connect 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 in...

Claims

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Application Information

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IPC IPC(8): A61K41/00A61K9/06A61K31/704A61K39/395A61K47/26A61K47/54A61P35/00
CPCA61K41/0052A61K31/704A61K47/26A61K47/549A61K9/06A61K9/0024A61K39/395A61P35/00A61K2039/55561A61K2300/00
Inventor 郑斌彭文畅明东刘爽甘霖
Owner TIANJIN UNIV
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