Superparamagnetism-modified and neutrophil exosome biomimetic vesicle-based biological preparation for drug delivery, and preparation method thereof

Patent metadata

Field	Value
Publication number	US-12544452-B2
Application number	US-202117787909-A
Country	US
Kind code	B2
Filing date	Apr 26, 2021
Priority date	Mar 26, 2021
Publication date	Feb 10, 2026
Grant date	Feb 10, 2026

Abstract

The present disclosure provides a superparamagnetism-modified and neutrophil exosome biomimetic vesicle-based biological preparation for drug delivery, and a preparation method thereof, belonging to the technical field of engineered nanovesicle drug loading systems. The superparamagnetism-modified and neutrophil exosome biomimetic vesicle-based biological preparation for drug delivery prepared by the preparation method acts on tumor cell lines. It is found that the superparamagnetism-modified and neutrophil exosome biomimetic vesicle-based biological preparation for drug delivery can specifically target tumor cells, induce tumor cell apoptosis, and inhibit tumor growth. The biological preparation significantly extends a survival time of mice and improves an efficacy of drugs against cancers.

First claim

What is claimed is:

1 . A preparation method of a superparamagnetism-modified and neutrophil exosome biomimetic vesicle-based biological preparation for drug delivery, comprising the following steps:
1) conducting co-culture on peripheral blood neutrophils with an antitumor drug to obtain neutrophils for uptake of the drug; 2) conducting centrifugation on the neutrophils for uptake of the drug, and resuspending a bottom cushion to obtain a cell suspension; 3) conducting continuous physical extrusion and gradient filtration on the cell suspension to obtain a neutrophil exosome biomimetic vesicle-encapsulated drug suspension; wherein the gradient filtration is conducted with polycarbonate filter membranes of 1 μm, 400 nm, and 200 nm sequentially; 4) conducting centrifugation on the neutrophil exosome biomimetic vesicle-encapsulated drug suspension, resuspending a bottom neutrophil nano vesicle (NNV)-drug cushion to obtain an NNV-drug suspension; and 5) mixing the NNV-drug suspension with a superparamagnetism material for co-incubation and conducting magnetic separation on a co-incubated mixture to obtain the superparamagnetism-modified and neutrophil exosome biomimetic vesicle-based biological preparation for drug delivery.

2 . The preparation method according to claim 1 , wherein in step 1), each 1×106 of the peripheral blood neutrophils are co-cultured with 50 μg of the antitumor drug.

3 . The preparation method according to claim 1 , wherein in step 1), the co-culture is conducted at 37° C. for 12 h to 24 h.

4 . The preparation method according to claim 1 , wherein in step 2), the centrifugation is conducted by differential centrifugation at 800 g for 5 min.

5 . The preparation method according to claim 1 , wherein in step (2), the bottom cushion is resuspended by phosphate-buffered saline (PBS).

6 . The preparation method according to claim 1 , wherein in step 3), the continuous physical extrusion is conducted 11 times on the cell suspension using a mini-extrader extruder.

7 . The preparation method according to claim 1 , wherein in step 3), each 5 mL of the cell suspension is subjected to the continuous physical extrusion at 300 psig with a temperature of a gas-tight syringe at not less than 10° C.

8 . The preparation method according to claim 1 , wherein in step 4), the centrifugation is conducted by ultracentrifugation at 10,000 g for 80 min.

9 . The preparation method according to claim 1 , wherein in step 4), the bottom NNV-drug cushion is resuspended by PBS.

10 . The preparation method according to claim 1 , wherein in step 5), the superparamagnetism material comprises Tf-SPION with a concentration in the NNV-drug suspension of 0.5 mg/ml.

11 . The preparation method according to claim 1 , wherein in step 5), the co-incubation is conducted at 4° C. for 4 h.

12 . The preparation method according to claim 1 , wherein in step 5), the magnetic separation is conducted by a neodymium magnet at a magnetic flux (MF) density of 1 T, and further wherein the antitumor drug comprises a doxorubicin (DOX).

13 . The preparation method according to claim 2 , wherein in step 1), the co-culture is conducted at 37° C. for 12 h to 24 h.

14 . The preparation method according to claim 5 , wherein in step (2), the bottom cushion is resuspended by phosphate-buffered saline (PBS).

15 . The preparation method according to claim 8 , wherein in step 3), each 5 mL of the cell suspension is subjected to the continuous physical extrusion at 300 psig with a temperature of a gas-tight syringe at not less than 10° C.

16 . The preparation method according to claim 11 , wherein in step 4), the bottom NNV-drug cushion is resuspended by PBS.

Assignees

• Univ Jiangsu

Inventors

• Zhang Xu
• Zhang Jiahui
• Ji Cheng
• Shi Hui
• Xu Wenrong
• Qian Hui

Classifications

• C12N2501/06

  Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide · CPC title

• C12N5/0642 Primary

  Granulocytes, e.g. basopils, eosinophils, neutrophils, mast cells · CPC title

• B03C2201/18

  Magnetic separation whereby the particles are suspended in a liquid · CPC title

• B03C1/02

  acting directly on the substance being separated · CPC title

• A61K47/52

  the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient · CPC title