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Neutrophile granulocyte drug-loading preparation and applications thereof

A neutrophil and drug-loading technology, applied in the direction of non-central analgesics, antipyretics, anti-inflammatory agents, etc., can solve the problem of reduced targeting efficiency, reduced targeting efficiency, and difficulty in screening out optimal structures, etc. problem, to achieve precise targeting and improve targeting efficiency

Inactive Publication Date: 2016-10-26
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the above-mentioned targeting vectors all have different limitations. For example, passive targeting vectors will be taken up by the liver and spleen after entering the body, reducing the targeting efficiency; active targeting vectors select the correct target site for high expression or specificity. Sexually expressed receptors or antigens are the primary key issue. Secondly, the usually selected small amount of low immunogenicity ligands such as: folic acid, growth factors, because they can be obtained from food, the human body itself has a very high concentration, so It will compete with the active targeting carrier to bind to the receptor, prevent the active delivery of the drug, and reduce the targeting efficiency; the structure design of the prodrug is difficult, and there are many kinds of hydrolytic enzymes or catalytic enzymes in the body, so it is difficult to screen out the optimal structure

Method used

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  • Neutrophile granulocyte drug-loading preparation and applications thereof
  • Neutrophile granulocyte drug-loading preparation and applications thereof
  • Neutrophile granulocyte drug-loading preparation and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Isolation of neutrophils: Draw out heparin with a 10mL syringe, and push out the heparin after wetting the wall of the syringe. 10 mL of venous blood was drawn from the mice. Take 2 mL of erythrocyte lysate (purchased from BD Company), mix it with venous blood, lyse the red blood cells at 4°C for 3 min, centrifuge at 1500 rpm for 3 min, and redisperse the pelleted cells with 1 mL of PBS (pH 7.4) to obtain a cell suspension. Take 0.6ml of 65% (V / V) cell separation solution (purchased from BD Company) in a centrifuge tube, add 0.3ml of 55% (V / V) cell separation solution on top, and finally add 0.3ml of cell suspension , 2500 rpm, centrifuged for 30 minutes, and collected the cell layer between the concentration of 65% cell separation liquid and the concentration of 55% cell separation liquid to obtain neutrophils. cells by 1.5 x 10 6 Each bottle was inoculated in a culture bottle containing RPMI-1640 medium, placed at 37°C, 5% CO 2 incubator culture, such as figure 1 ...

Embodiment 2

[0027] Preparation of antineoplastic drug-loaded cells: the neutrophils prepared in Example 1 were mixed with 1×10 6 cells / well were inoculated in 24-well cell culture plates (containing RPMI-1640 medium), placed at 37°C, 5% CO 2 After pre-incubating in the incubator for 0.5 h, the medium was aspirated, and 100 μL of paclitaxel albumin nanoparticles (purchased from Abraxis BioScience) at a concentration of 2 mg / mL was added to each well for 2 h, and washed 3 times with RPMI-1640 medium preheated at 37 °C. Scrape the cells with a cell scraper and centrifuge at 1200rpm for 10min. Discard the supernatant, add 10mL PBS, repeat 3 times, collect the drug-loaded cells, which is the neutrophil drug-loaded preparation; set aside.

Embodiment 3

[0029] Preparation of antineoplastic drug-loaded cells: the neutrophils prepared in Example 1 were mixed with 1×10 6 cells / well were inoculated in 24-well cell culture plates (containing RPMI-1640 medium), placed at 37°C, 5% CO 2 After pre-incubating in the incubator for 0.5 h, suck out the medium, add 100 μL of 2 mg / mL doxorubicin liposome (purchased from Roche MabThera) to each well and incubate for 2 h, wash with RPMI-1640 medium preheated at 37 °C for 3 times , scrape the cells with a cell scraper, and centrifuge at 1200rpm for 10min. Discard the supernatant, add 10mL PBS, repeat 3 times, collect the drug-loaded cells, measure the concentration, and set aside.

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Abstract

The invention discloses a neutrophile granulocyte drug-loading preparation and applications of the neutrophile granulocyte drug-loading preparation. According to the neutrophile granulocyte drug-loading preparation, neutrophile granulocytes are taken as carriers, to-be-loaded drugs enter the carriers under the effect of receptor-mediated endocytosis, macropinocytosis or phagocytosis, and thus the neutrophile granulocyte drug-loading preparation is obtained; aiming at the defects in the existing targeting preparations, the neutrophile granulocyte drug-loading preparation has the advantages that rat neutrophile granulocytes and the drugs are co-incubated, so that the drug-loading cells are formed and then used for the in-vivo drug delivery, and finally, the targeting efficiency is improved.

Description

(1) Technical field [0001] The invention relates to a targeted drug delivery preparation, in particular to a preparation and application using neutrophils as drug carriers. (2) Background technology [0002] Targeted drug delivery system (Targeting drug delivery system) belongs to the drug controlled release system, which involves many fields such as pharmacy, biology, chemistry, chemical industry, materials science, etc. It refers to the concentration of drugs through local or systemic blood circulation The drug delivery system is located in the target tissue, target organ and target cell. Targeted drug delivery system is also a kind of drug carrier system, which has the ability to selectively transport and release drugs to target tissues, target organs or target cells, increase the drug concentration in the target area, reduce the concentration in other non-target parts, and reduce toxic and side effects. characteristic. Targeted drug delivery systems have been developed...

Claims

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Application Information

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IPC IPC(8): A61K47/42A61K31/337A61K31/704A61K31/573A61K9/14A61K9/127A61P35/00A61P25/00A61P29/00
CPCA61K47/42A61K9/127A61K9/146A61K31/337A61K31/573A61K31/704
Inventor 靳翔
Owner ZHEJIANG UNIV OF TECH
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