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Antiacid nano oral deoxyribonucleic acid (DNA) anti-tumor vaccine with potential of hydrogen (pH) sensitive characteristic and preparation method

A sensitive and anti-tumor technology, which is applied in the direction of anti-tumor drugs, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc. It can solve the problems of low expression efficiency, large toxic and side effects of nanoparticle carriers, and oral administration. Pathway instability and other issues, to achieve strong antigen presentation, facilitate phagocytosis, and reduce the effect of inhibition

Inactive Publication Date: 2012-08-01
NANKAI UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problems that the nanoparticle carriers currently used in experimental tumor immunotherapy have high toxicity and side effects, unstable oral administration route, low expression efficiency and difficulty in carrying DNA through the gastric acid environment and entering the intestinal tract to efficiently express tumor antigens. A novel acid-resistant nanoparticle oral DNA antitumor vaccine with pH-sensitive properties

Method used

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  • Antiacid nano oral deoxyribonucleic acid (DNA) anti-tumor vaccine with potential of hydrogen (pH) sensitive characteristic and preparation method
  • Antiacid nano oral deoxyribonucleic acid (DNA) anti-tumor vaccine with potential of hydrogen (pH) sensitive characteristic and preparation method
  • Antiacid nano oral deoxyribonucleic acid (DNA) anti-tumor vaccine with potential of hydrogen (pH) sensitive characteristic and preparation method

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Experimental program
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Effect test

preparation example Construction

[0034] 1. Preparation of Chitosan Nanoparticles

[0035] 1.1. Dissolve long-chain chitosan with a molecular weight of 28000 to 32000 in double-distilled water, adjust the pH to 5.2, and stir at 55°C for 30 minutes to form liquid A;

[0036] Section 1.2, at the same time 25mM Na 2 SO 4 Mix the solution with the required DNA plasmid and heat to 55°C, stir at a constant speed for 30 minutes to form liquid B;

[0037] 1.3. Then mix the two solutions A and B in a volume ratio of 1:1, and mix them immediately, and store the prepared chitosan nanoparticles at 4°C after 5 minutes;

[0038] 2. Activation of the carboxyl group on the side chain of alginic acid

[0039] Dissolve the alginic acid powder in double distilled water, and mix the alginic acid solution with the added N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. Mix with a ratio of 1:5:5, adjust the pH to 9.0, stir overnight at room temperature, and store the activated alginic acid at 4°C for late...

Embodiment 1

[0062] 1. Preparation of acid-resistant nanoparticle oral DNA anti-tumor vaccine with pH sensitivity:

[0063] (1) Preparation of chitosan nanoparticles: Dissolve long-chain chitosan with a molecular weight of 28,000 to 32,000 in double distilled water, adjust the pH to 5.2, and stir at 55°C for 30 minutes.

[0064] (2) At the same time, add 25mM Na 2 SO 4 The solution was mixed with Legumain DNA plasmid and heated to 55°C and stirred at a constant speed for 30 minutes. Then mix the two solutions at a volume ratio of 1:1, and mix them immediately, and store the prepared chitosan nanoparticles at 4°C after 5 minutes;

[0065] (3) Activation of the carboxyl group on the side chain of alginic acid: dissolve the alginic acid powder in double distilled water, and mix the alginic acid solution with the added N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)- 3-ethylcarbodiimide hydrochloride was mixed at a molar ratio of 1:5:5, and the pH was adjusted to 9.0, stirred overnight at room te...

Embodiment 2

[0071] The pH-sensitive acid-resistant nanoparticle oral DNA anti-tumor vaccine expresses the encoded protein in the body of normal BALB / c mice.

[0072] (1) Gavage and administration dose: each BALB / c mouse is given 150μg DNA plasmid with acid-resistant nanoparticles oral DNA anti-tumor vaccine or 10 8 Salmonella in CFU, once a day;

[0073] (2) Repeat the above steps 5 times;

[0074] (3) In vivo fluorescence expression detection of BALB / c tumor-bearing mice after oral immunization: the tumor-bearing mice were sacrificed, and the small intestinal Peyer's lymph nodes were taken to observe the oral DNA of acid-resistant nanoparticles encoding the pH-sensitive properties of green fluorescent protein The expression of anti-tumor vaccine and compare it with other controls.

[0075] The result is Figure 7 As shown, green fluorescent protein is highly expressed in macrophages in mouse Peyer's lymph nodes. Figure 8 The results of flow cytometry analysis showed that the expression of green...

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Abstract

Provided are an antiacid nano particle oral deoxyribonucleic acid (DNA) anti-tumor vaccine with the potential of hydrogen (pH) sensitive characteristic and a preparation method. The DNA oral vaccine is nano particles formed by combining chitosan with alginic acid for surface finish and encoded tumor specific antigen Legumain protein DNA plasmids. The DNA oral vaccine is capable of being efficiently phagocytized by dendritic cells and macrophage in peyer's patches and expressing the encoded tumor antigen to activate immune damage of a host for tumor cells. The nano particles have small toxic and side effects and a strong antigen presentation role. Specifically, the DNA plasmids and chitosan are led to be in static negative crosslinking, and then, alginic acid is used for surface finish of chitosan nano particles. By means of immunofluorescent staining and flow cytometry, growth and transfer conditions of breast cancer of a mouse are estimated, immune response of the mouse is analyzed, and antigen presentation capability and anti-tumor effect of the antiacid nano particle oral DNA anti-tumor vaccine with the potential of pH sensitive characteristic are determined.

Description

Technical field [0001] The invention relates to a novel acid-resistant nanoparticle oral DNA anti-tumor vaccine with pH-sensitive characteristics, and a treatment method using the same. It belongs to the field of genetic engineering pharmaceutical technology. Background technique [0002] Breast cancer has become a common tumor in women. According to the National Cancer Institute (NCI) statistics, from 2004 to 2008, the average age of patients diagnosed with breast cancer was 61 years old, and the annual incidence rate among women was about 0.0122%, of which Asian / Pacific Island The incidence rate of ethnicity is about 0.0937%; from 2003 to 2007, the average age of death of breast cancer patients was 68 years old, and the annual mortality rate among women was 0.024%, of which the annual mortality rate of Asian / Pacific Islanders was 0.0122%; Rate and death rate are the first and second place among women. The current treatment of breast cancer is mainly traditional therapies, na...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/00A61K9/10A61K47/36A61P35/00
Inventor 刘赜谭小月刘淑熊敏吕丹向荣
Owner NANKAI UNIV
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