Anti-tumor immunotherapy nano-drug delivery system and construction method thereof

A technology of anti-tumor immunity and drug delivery system, which is applied in the field of anti-tumor immunotherapy nano drug delivery system and its construction, and drug delivery system. It can solve the problems of poor anti-tumor DNA or peptide vaccine effects, and achieve or reverse tumor immunity. Escape, inhibit tumor immune escape, increase the effect of anti-tumor immune response

Active Publication Date: 2016-03-16
ZHEJIANG PHARMA COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the above-mentioned technical problem of poor in vivo effect of existing anti-tumor DNA or polypeptide vaccines, the present invention provides an anti-tumor immunotherapeutic nanotechnology that can not only increase the anti-tumor immune response of tumor-bearing organisms, but also inhibit or reverse tumor immune escape. The drug delivery system and its construction method have the advantages of simple preparation method and high targeting efficiency

Method used

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  • Anti-tumor immunotherapy nano-drug delivery system and construction method thereof
  • Anti-tumor immunotherapy nano-drug delivery system and construction method thereof
  • Anti-tumor immunotherapy nano-drug delivery system and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of paclitaxel / anionic cyclodextrin-DNA / mannose modified trimethyl chitosan nanocomposite. The specific steps are as follows:

[0038] 1.1 Preparation of paclitaxel / anionic cyclodextrin inclusion compound (hereinafter referred to as PTX / SBE inclusion compound), paclitaxel (Paclitaxel, hereinafter referred to as PTX) was added to 0.25 mg·mL at a molar ratio of 1:1 -1 Anionic cyclodextrin (Sulfobutylether-β-Cyclodextrin, hereinafter referred to as SBE), at 25 ℃ temperature and 1000w power for 4h, then centrifuged at 10000g for 10min, and finally filtered through 0.45μm microporous membrane, A PTX / SBE clathrate was obtained.

[0039] 1.2 Construction of pEGFP-N2-Trp2-GM-CSF-Fc plasmid.

[0040] (1) Design primers according to the functional coding region sequences of Trp2, GM-CSF, and Fc in GenBank, extract total RNA from human peripheral leukocytes or lymphocytes, and synthesize cDNA, using the first cDNA that was reverse-transcribed as a template,...

Embodiment 2

[0046] Example 2: Structural characterization of the PTX / SBE-DNA / Man-TMC nanocomposite.

[0047] 2.1 Select an appropriate amount of PTX / SBE-DNA / Man-TMC nanocomposite, add ultrapure water to dilute, use a laser particle size analyzer to measure the particle size and Zeta potential value of the PTX / SBE-DNA / Man-TMC nanocomposite, Each experiment was repeated three times, and the average value was used for statistical analysis. Please also refer to figure 2 , image 3 and Table 1, figure 2 It is a histogram of paclitaxel / anionic cyclodextrin-DNA / mannose modified trimethyl chitosan nanocomposite nanoparticle particle size provided by the present invention; image 3 It is the distribution diagram of Zeta voltage curve of paclitaxel / anionic cyclodextrin-DNA / mannose modified trimethyl chitosan nanocomposite nanoparticle provided by the present invention. Depend on figure 2 And table 1 can observe that the particle diameter of described PTX / SBE-DNA / Man-TMC nanocomposite nanopa...

Embodiment 3

[0051] Example 3: Determination of the effect of the PTX / SBE-DNA / Man-TMC nanocomposite on dendritic cells (Dendritic cells, hereinafter referred to as DC).

[0052] 3.1 To investigate the effect of nanocomposites on the phenotype of dendritic cells.

[0053] The DC immature dendritic cells on the 7th day of culture were collected, washed once with PBS, and seeded in 6-well cell culture plates (1×10 6 / well), add SBE-DNA / Man-TMC, PTX / SBE-DNA / Man-TMC, PTX / SBE-DNA / TMC respectively, the content of DNA in the total system is 1.135μg·mL -1 , with lipopolysaccharides (Lipopolysaccharides, hereinafter referred to as LPS) stimulated cells as a control (670ng·mL -1 ). Incubate at 37°C for 48 hours, wash with PBS twice, add PE-labeled anti-mouse CD11c, FITC-labeled anti-mouse CD80, PE-Cy5-labeled anti-mouse CD86 each 2 μL, and then incubate at 4°C 1h, and under the same operation, label each control cell sample with the isotype control antibody of each mouse anti-antibody to eliminate...

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Abstract

The invention discloses an anti-tumor immunotherapy nano-drug delivery system. The anti-tumor immunotherapy nano-drug delivery system consists of a taxol / anionic cyclodextrin-DNA / mannose modified trimethyl chitosan nano-composite; the anti-tumor immunotherapy nano-drug delivery system is composed of taxol, anionic cyclodextrin, DNA and mannose modified trimethyl chitosan; and the anti-tumor immunotherapy nano-drug delivery system is targeted to dendritic cells by virtue of a mannose ligand. The invention also discloses a construction method of the anti-tumor immunotherapy nano-drug delivery system. The comprehensive immune drug delivery system provided by the invention not only can enhance the antitumor immunity response of a tumor bearing body but also can inhibit or reverse tumor immune escape; and the nano-drug delivery system is simple and convenient in preparation method and high in targeting efficiency.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a drug delivery system, in particular to a nano drug delivery system for anti-tumor immunotherapy and a construction method thereof. Background technique [0002] Tumor immunotherapy is a new tumor treatment method following surgery, drug therapy, and radiation therapy. It has unique advantages in preventing tumor recurrence, metastasis, and treating tumor lesions. The world's first tumor therapeutic vaccine, the epidermal growth factor (EGF) tumor therapeutic vaccine, was launched in June 2008. The vaccine can effectively prolong the survival period of patients with advanced lung cancer. The side effects are very mild and disappear automatically after stopping the drug, and no symptoms of autoimmune reactions occur. It can be seen that tumor therapeutic vaccines have obvious advantages in anti-tumor therapy. Existing research on anti-tumor immunotherapy mainly focuses on improving th...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K9/14A61K39/39A61K39/00A61P35/00
Inventor 胡英许娇娇
Owner ZHEJIANG PHARMA COLLEGE
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