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Co-drug-loading cell microparticle preparation and preparation method thereof

A technology of micro-particles and co-loading drugs, which is applied in the fields of pharmaceutical formulations, anti-tumor drugs, micro-capsules, etc., can solve the problems of ineffective enrichment and penetration, insufficient anti-tumor immune activation level, and limited photothermal treatment effect. To achieve the effect of weakening the ability of antigen presentation, improving the tumor microenvironment, and inhibiting the death of tumor-specific killer T cells

Active Publication Date: 2022-08-09
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the prior art, the present invention can effectively solve the problems that photosensitizers and tumor-associated fibroblast deactivators cannot be effectively enriched and penetrated in tumor tissues, the effect of photothermal therapy is limited, and the level of anti-tumor immune activation is insufficient.

Method used

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  • Co-drug-loading cell microparticle preparation and preparation method thereof
  • Co-drug-loading cell microparticle preparation and preparation method thereof
  • Co-drug-loading cell microparticle preparation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Preparation and characterization of co-loaded ICG / Cal cell microparticles

[0055] 1. Experimental materials and reagents

[0056] H22 mouse hepatoma cells, indocyanine green (ICG), calcipotriol (Cal), and the ultraviolet device is owned by the conventional cell ultra-clean workbench.

[0057] 2. Experimental steps

[0058] 1) H22 mouse liver cancer cells were irradiated with ultraviolet light for 1 h, and 100 μg ml was added to the cells -1 ICG and 4 μg ml - 1 Cal, placed in a 37°C incubator.

[0059]2) After co-incubating for 12 h, apoptotic tumor cells were separated. Take the supernatant and centrifuge step by step, centrifuge at 200g and 600g for 10min each, and then centrifuge at 14,000g for 1min to remove cells and debris. Take the centrifuged supernatant and further centrifuge at 20,000g for 1h to obtain H22 mice Co-encapsulated cell drug-loaded microparticles (Cal / ICG@MPs) produced by apoptosis of liver cancer cells were used as the experimenta...

Embodiment 2

[0063] Example 2: Photothermal killing effect of co-loaded ICG / Cal cell microparticles on H22 mouse hepatoma cells

[0064] 1. Experimental materials and reagents

[0065] H22 mouse hepatoma cells, indocyanine green (ICG), calcipotriol (Cal) and flow-through antibodies were obtained commercially.

[0066] 2. Experimental steps

[0067] 1) The collection of microparticles co-loaded with ICG / Cal cells is the same as in Example 1.

[0068] 2) To 1*10 4 H22 mouse hepatoma cells were added with (4μg / mL, ICG amount; 60ng / mL, Cal amount) free ICG, Cal / ICG (that is, with both free Cal and free ICG), ICG@MPs, Cal / ICG@MPs The serum-free medium was incubated at 37°C under 5% CO2 for 4 h, the cells were collected, washed three times with PBS, and centrifuged at 250 g for 3 min. Using 808nm laser (1W / cm 2 , 5 min) treated H22 mouse hepatoma cells. After incubation at 37°C and 5% CO2 for 4 hours, 10 μL of CCK8 solution was added to each well and incubated for 4 hours. A 318C microplat...

Embodiment 3

[0072] Example 3: Photothermal killing effect of co-loaded ICG / Cal cell microparticles on 4T1 mouse breast cancer cells

[0073] 1. Experimental materials and reagents

[0074] 4T1 mouse breast cancer cells, indocyanine green (ICG), calcipotriol (Cal) and flow-through antibodies were obtained commercially.

[0075] 2. Experimental steps

[0076] 1) The collection of co-loaded ICG / Cal cell microparticles is the same as in Example 1, wherein the tumor cell microparticles are derived from 4T1 mouse breast cancer cells.

[0077] 2) The photothermal treatment and CRT detection methods of 4T1 mouse breast cancer cells are the same as those in Example 2. Cal / ICG@MPs were added to 4T1 mouse breast cancer cells and treated with laser as the experimental group. Add (4μg / mL, ICG amount) free ICG and laser treatment as control group 3, add (4μg / mL, ICG amount; 60ng / mL, Cal amount) free Cal / ICG and laser treatment as a control In group 4, ICG@MPs containing (4 μg / mL, ICG amount) and la...

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Abstract

The invention belongs to the technical field of drug targeting carriers, and discloses a co-drug-loading cell microparticle preparation and a preparation method thereof, the co-drug-loading cell microparticle preparation comprises microparticles secreted from tumor cells, and a photosensitizer and a tumor-associated fibroblast deactivator together entrapped in the microparticles. The detailed composition and the structure of the preparation are improved, microparticles secreted from tumor cells are taken as carriers, and the photosensitizer and the tumor-related fibroblast deactivator are simultaneously entrapped in the carriers, so that the obtained co-drug-loading cell microparticle preparation has the advantages that on one hand, the tumor matrix microenvironment is transformed, the enrichment of the photosensitizer in tumors is enhanced, and the tumor-related fibroblast deactivator can be effectively inhibited; on the one hand, the direct killing effect of photothermal therapy on tumor cells can be improved, on the other hand, tumor immunogenic death caused by photothermal therapy can be enhanced, CD8 + T cell infiltration can be promoted, CD8 + T cell death induced by antigen-dependent activation caused by tumor-related fibroblasts can be reduced, and the tumor immune microenvironment can be improved.

Description

technical field [0001] The invention belongs to the technical field of drug targeting carriers, and more particularly, relates to a co-drug-loaded cell microparticle preparation and a preparation method thereof. The package contains two components, a photosensitizer and a tumor-associated fibroblast deactivator, which is especially suitable for anti-tumor applications. Background technique [0002] Despite the tremendous achievements in cancer research, cancer is still one of the leading causes of death, and more effective tumor treatment strategies need to be further explored. Photothermal therapy (PTT) is a tumor treatment method that has been widely developed in recent years. Under laser irradiation, light energy is converted into heat energy through photosensitizers, which can provide a spatiotemporal thermal effect to ablate tumor tissue in the light-irradiated area. Special treatment effect, and no damage to normal tissue during treatment. However, the dense matrix m...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K47/46A61K41/00A61K45/06A61K31/593A61P35/00
CPCA61K41/0057A61K45/06A61K41/0052A61K9/5068A61P35/00A61K31/593A61K41/0071A61K2300/00Y02A50/30
Inventor 甘璐雍土莹李新杨祥良
Owner HUAZHONG UNIV OF SCI & TECH
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