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Macrophage vesicle entrapped nano-drug preparation and application thereof in treating arthritis

A technology of macrophages and nano-medicines, applied in the field of biotechnology, which can solve the problems of complex and low efficiency of cell membrane extraction methods

Inactive Publication Date: 2020-06-30
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the most widely used biomimetic strategy is to use the cell membrane to encapsulate the drug-loading system and endow the drug-loading system with specific biological functions (Li R, He Y, Zhang S, et al. Cell membrane-based nanoparticles: a new biomimetic platform for tumor diagnosis and treatment.Acta PharmaceuticaSinica B. 2018; 8:14-22.); therefore, a large amount of efficient macrophage membrane extraction is crucial to the construction of a biomimetic drug delivery system targeting RA; however, traditional cell membrane extraction methods are complex and efficient Aiming at the above problems, a feasible method is to use cell vesicles to replace the cell membrane-encapsulated drug-carrying system

Method used

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  • Macrophage vesicle entrapped nano-drug preparation and application thereof in treating arthritis
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  • Macrophage vesicle entrapped nano-drug preparation and application thereof in treating arthritis

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1: Extraction and Characterization of Macrophage Vesicles

[0042] In this example, the macrophage RAW264.7 was first cultured, and then the macrophage vesicles were efficiently extracted by drug stimulation. The specific operation is as follows: culture RAW264.7 cells in complete DMEM medium, add 10 μg / mL cytochalasin B, incubate at 37 degrees for 1 hour, remove the medium, add 5 mL of serum-free medium to collect cells by pipetting, and vortex for 5 minutes Add 5 mL of serum, centrifuge at 1000 rpm for 5 min, remove cells, collect supernatant and centrifuge at 4000 rpm for 15 min to obtain macrophage vesicle MMV. Rinse twice with 0.25% EDTA solution to remove the vesicle content to obtain the macrophage vesicle membrane ( figure 1 B). Then by BCA quantification, the macrophage vesicle protein content was analyzed, and it was found that the MMV membrane protein content was much higher than that of the macrophage membrane extracted by traditional methods ( fi...

Embodiment 2

[0043] Example 2: Nanoparticles Encapsulated by Macrophage Vesicles

[0044] In this example, the common degradable biomaterial PLGA is used. The preparation method of PLGA nanoparticles (T-NPs) loaded with tacrolimus is the nanoprecipitation method. The specific operation is as follows: Weigh an appropriate amount of PLGA (0.67dL / g , 50:50, carboxyl-terminated) was dissolved in acetone to prepare a PLGA stock solution with a concentration of 10 mg / ml. And add 10% (w / w) Tacrolimus, dissolve completely. Add 1ml of pure water to the vial, quickly inject 0.5ml of Tacrolimus-containing PLGA acetone solution, and place the acetone in a fume hood to completely evaporate the acetone to obtain a 5mg / ml Tacrolimus PLGA nanoparticle (T-NP) solution. Store in refrigerator at 4°C until use. In order to screen the optimal prescription, different dosages of Tacrolimus were added, and it was found that precipitation would occur if the dosage exceeded 15% ( image 3 B).

[0045] Take the ...

Embodiment 3

[0046] Example 3: Characterization of Macrophage Vesicle-Encapsulated Nanoparticles

[0047] Observation under the transmission electron microscope after negative staining with uranyl acetate, the MNP is regular spherical, with obvious core-shell structure, uniform size and good dispersion ( figure 2 A). Potential / laser particle size analyzer measurement results show that the MNP average particle size is 130nm ( figure 2 B), about 30nm larger than the PLGA nanoparticle core, consistent with cell membrane thickness. , the potential is -25mV consistent with the MMV potential. The encapsulation efficiency and drug loading capacity of the nanoparticles are determined by centrifuging to measure the drug content in the nano-precipitation, the encapsulation efficiency is 98.5%, and the drug loading capacity is 9.1%.

[0048] Stability investigation of MNP. Disperse MNP in PBS, measure and record the average particle size of MNP every day, and measure continuously for one week. ...

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Abstract

The invention belongs to the technical field of biology, relates to preparation of bionic nano-carriers and targeted treatment of arthritis, and particularly relates to a macrophage vesicle entrappednano-drug preparation and application thereof in treating arthritis. The nano-drug preparation targets an arthritis part through adhesive protein on the surface of a macrophage vesicle, and an immunosuppressive drug is delivered for targeted treatment of arthritis. Furthermore, a new strategy for targeted treatment of arthritis is provided for clinical practice, and the strategy comprises the steps of fusing an artificial drug carrier and the natural macrophage vesicle, and constructing the stable macrophage vesicle entrapped nano-drug preparation, thereby improving the targeted treatment effect of arthritis.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to the preparation of bionic nano-carriers and targeted therapeutic drugs for arthritis, in particular to a nano-medicine preparation carried by macrophage vesicles and its use in the preparation of drugs for treating arthritis, in particular to a Extraction of macrophage vesicles with arthritis-targeting performance, biomimetic nano-medicine preparation and application thereof, the biomimetic nano-medicine targets arthritis sites through adhesion proteins on the surface of macrophage vesicles, and delivers Immunosuppressive drugs, thereby enhancing the efficacy of arthritis-targeted therapies. Background technique [0002] Rheumatoid arthritis (RA) is a relatively common autoimmune disease. Because of its high disability rate and serious complications, RA seriously threatens human health. Its main manifestations are chronic progressive polyarticular inflammation; damage to the s...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/46A61K47/34A61K31/436A61P29/00A61P19/02
CPCA61K9/5176A61K9/5153A61K31/436A61P29/00A61P19/02
Inventor 王建新庞志清李瑞翔何雨薇
Owner FUDAN UNIV
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