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Neutrophil-like nano drug delivery system as well as preparation method and application thereof

A nano-drug delivery system and neutrophil technology, applied in antibacterial drugs, pharmaceutical formulations, nervous system diseases, etc., can solve the problems of hidden safety hazards, low drug loading, and high overall cost, and reduce toxic and side effects. Simple method and controllable effect

Pending Publication Date: 2022-01-21
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cell carriers have high requirements in preparation, drug loading, storage and other links, and their drug loading is relatively low, and the overall cost is high
In addition, since the cell carrier still has certain activity, there may be certain safety hazards in the circulation in the body

Method used

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  • Neutrophil-like nano drug delivery system as well as preparation method and application thereof
  • Neutrophil-like nano drug delivery system as well as preparation method and application thereof
  • Neutrophil-like nano drug delivery system as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Preparation and collection of activated neutrophils

[0040] 1) Under sterile conditions, ICR mice (6 weeks old) were sacrificed by decapitation, and their femurs and tibias were isolated. Afterwards, the bone marrow cavity was exposed, and fresh RPMI.1640 culture solution was aspirated with a disposable sterile syringe with a needle, and the bone marrow cavity was washed repeatedly until the bone was white and translucent. After washing, the washing solution was filtered through a 200-mesh cell sieve and transferred to a centrifuge tube, and the cells were collected by centrifugation (1700 rpm, 5 min).

[0041] 2) Add 3 to 5 times the volume of erythrocyte lysate, centrifuge to collect cells after lysing for 1 to 2 minutes, and centrifuge with a certain volume of phosphate buffer (1700rpm, 3min) to wash twice, and finally add 2mL PBS to resuspend to make a cell suspension . Neutrophils were separated and purified by Percoll discontinuous density gradient c...

Embodiment 2

[0043] Example 2: Preparation and collection of activated neutrophils

[0044] 1) Under sterile conditions, blood was collected through the ophthalmic vein plexus, and the mouse blood was collected with an anticoagulant tube containing EDTA. Subsequently, the whole blood was centrifuged (3220g, 5min, 4°C), and the middle buffy coat was taken and diluted to 2mL with PBS.

[0045] 2) Add 3 to 5 times the volume of erythrocyte lysate, centrifuge to collect cells after lysing for 1 to 2 minutes, and centrifuge with a certain volume of phosphate buffer (1700rpm, 3min) to wash twice, and finally add 2mL PBS to resuspend to make a cell suspension . Neutrophils were separated and purified by Percoll discontinuous density gradient centrifugation. Dilute the Percoll stock solution with 1.5M NaCl at a volume ratio of 9:1 to obtain 100% Percoll separation solution, and dilute with 0.15M NaCl in proportion to obtain 78%, 65% and 55% Percoll separation solution, respectively. Take a 15mL...

Embodiment 3

[0047] Example 3: Preparation and collection of activated neutrophils

[0048] 1) Under sterile conditions, ICR mice (6 weeks old) were sacrificed by decapitation, and their femurs and tibias were isolated. Afterwards, the bone marrow cavity was exposed, and fresh RPMI.1640 culture solution was aspirated with a disposable sterile syringe with a needle, and the bone marrow cavity was washed repeatedly until the bone was white and translucent. After washing, the washing solution was filtered through a 200-mesh cell sieve and transferred to a centrifuge tube, and the cells were collected by centrifugation (1700 rpm, 5 min).

[0049] 2) Add 3 to 5 times the volume of erythrocyte lysate, centrifuge to collect cells after lysing for 1 to 2 minutes, and centrifuge with a certain volume of phosphate buffer (1700rpm, 3min) to wash twice, and finally add 2mL PBS to resuspend to make a cell suspension . Neutrophils were separated and purified by Percoll discontinuous density gradient c...

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Abstract

The invention discloses a neutrophil-like nano drug delivery system, which has a phospholipid bilayer structure and is prepared by interactive fusion of an activated neutrophil membrane stimulated by a stimulating factor and phospholipid molecules, and the particle size of the nano drug delivery system is 100-300nm. The invention further discloses a preparation method and application of the neutrophil-like nano drug delivery system. According to the neutrophil-like nano drug delivery system with the membrane-membrane fusion structure, the advantages of the liposome and the cell carrier are fully combined, the respective defects of the liposome and the cell carrier are avoided, the liposome is used for loading drugs to overcome the defect that the cell carrier is difficult to load drugs, the active targeting ability of the liposome is endowed by the activated-state neutrophil membrane, the combination of the liposome and the cell carrier is beneficial to realizing accurate delivery of the drug to the inflammation part, can improve the treatment effect, can reduce the toxic and side effects, and has broad application prospects.

Description

technical field [0001] The invention relates to the field of cell bionic drug delivery system construction, in particular to a neutrophil-like nano drug delivery system and its preparation method and application. Background technique [0002] Inflammation-related diseases, such as malignant tumors and bacterial infections, are an important factor threatening human health. For the treatment of inflammation-related diseases, systemic drug therapy is still the main clinical practice. However, due to the lack of specific identification of lesions, systemic drug therapy is more likely to cause systemic side effects; and if the concentration of drugs distributed in the lesions is lower than the therapeutic limit, it is easy to cause drug resistance, which will affect malignant tumors. The follow-up treatment of inflammation-related diseases represented by bacterial infections and bacterial infections has caused great difficulties. Therefore, realizing the effective delivery of d...

Claims

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Application Information

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IPC IPC(8): A61K47/24A61K47/46A61K41/00A61P35/00A61P31/04A61P35/04A61P17/02A61P3/10A61P19/02A61P25/00A61K49/00
CPCA61K47/24A61K47/46A61K41/0057A61P35/00A61P31/04A61P35/04A61P17/02A61P3/10A61P19/02A61P25/00A61K49/0034
Inventor 高建青吴佳禾林能明
Owner ZHEJIANG UNIV
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