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Tumor antigen presentation system and applications of the same in preparation antineoplastic drugs, and immune system activator and antineoplastic composition

A technology of anti-tumor drugs and tumor antigens, which can be used in anti-tumor drugs, drug combinations, blood/immune system cells, etc., and can solve problems such as high safety

Active Publication Date: 2019-05-07
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, when the antigen presentation system modified or associated with chemical substances is used in humans, the safety of its use is the biggest problem

Method used

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  • Tumor antigen presentation system and applications of the same in preparation antineoplastic drugs, and immune system activator and antineoplastic composition
  • Tumor antigen presentation system and applications of the same in preparation antineoplastic drugs, and immune system activator and antineoplastic composition
  • Tumor antigen presentation system and applications of the same in preparation antineoplastic drugs, and immune system activator and antineoplastic composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Preparation of tumor antigen presentation system

[0048] Blood was removed from the orbit of C57BL / 6 mice and stored in PBS containing 1% EDTA. Centrifuge at 2000rpm for 5 minutes, remove the supernatant, and repeat the centrifugation step until pure red blood cells are obtained. Add deionized water containing 1% EDTA to the erythrocytes, shake gently, and centrifuge the mixture at 4000 rpm for 10 minutes after 5 minutes. Separate the precipitate from the supernatant, add deionized water containing 1% EDTA to the precipitate, sonicate at a frequency of 40kHZ for 30 seconds, add ice cubes to the water bath to cool down during the sonication process, and then centrifuge at 14800rpm at 4°C 20 minutes. After pouring out the supernatant, add deionized water containing 1% EDTA to the precipitate, then perform ultrasonic treatment for 30 seconds according to the above conditions, and further centrifuge at 14800rpm and 4°C for 20 minutes, and repeat the above cent...

Embodiment 2

[0049] Example 2: Extraction of tumor cell membrane

[0050] Extract the cell membrane of melanoma cell B16F10:

[0051] Melanoma cells B16F10 were resuspended in cell disruption solution (containing 0.25M sucrose, 1mM EDTA, 20mM Hepes-NaOH and protease inhibitors, pH 7.4) and sonicated on ice for 30 cycles with a Selecta Sonopuls cell sonicator (each cycle Ultrasound for 3 seconds, pause for 7 seconds), the resulting mixture was centrifuged at 4000rpm, and the supernatant was taken out and further centrifuged at 14800rpm at 4°C for 20 minutes to obtain pure cell membranes of melanoma cells B16F10, which were resuspended in PBS.

[0052] Extraction of tumor cell membrane in tumor tissue:

[0053] After the B16-luc tumor tissue was sliced, it was sonicated on ice for 30 cycles using a Selecta Sonopuls cell sonicator (3 seconds per cycle, 7 seconds pause). The B16-luc tumor tissue was centrifuged at 6000rpm for 10 minutes to remove the pigment, and then the supernatant was co...

Embodiment 3

[0054] Example 3: Fusion of tumor antigen presentation system and tumor cell membrane (preparation of immune system activator)

[0055] Measure the protein content of the erythrocyte membrane and the tumor cell membrane that obtain in embodiment 1 and embodiment 2 with BCA kit, and according to the proportion (20:1,10:1,5:1 respectively) of erythrocyte membrane protein than tumor cell membrane protein ratio , 2:1, 1:1, and 0:1) for mixing, ultrasonication in a water bath for 15 minutes until the system became transparent, and ice cubes were added during the ultrasonication process to prevent protein inactivation caused by excessive system temperature. The system was then shaken in a metal bath at 37°C for 30 minutes, this step was to promote the process of membrane fusion. Finally, the mixed membrane was sonicated in a water-bath ultrasonicator for 2 minutes, and repeatedly filtered with a Millipore 450nm syringe filter to finally obtain nanoplasmic membrane vesicles with fusi...

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Abstract

The invention discloses a tumor antigen presentation system and applications of the same in preparation antineoplastic drugs. The system is prepared by erythrocyte; nano-plasmalemma vesicles of fusionmembranes can be formed through ultrasound fusion and extrusion of the system and tumor cell membranes, and can effectively activate immune systems; and the system is derived from the organism itself, so the source is safe, and no chemical modification reagents are added during preparation, and therefore, using safety can be further ensured. An antineoplastic composition is also disclosed. The antineoplastic composition includes the nanomembrane vesicles of erythrocyte membranes-tumor cell membrane fusion membranes, and an immune checkpoint inhibitor; the antineoplastic composition can effectively enhance the activity of DC cells, NK cells, B cells and T cells, and can effectively inhibit the growth of tumor cells; and in addition, the antineoplastic composition can effectively reduce therelapse and metastasis of tumors.

Description

technical field [0001] The invention relates to a tumor antigen presentation system and its application in the preparation of antitumor drugs, an immune system activator and an antitumor composition. Background technique [0002] Cancer is the most common type of malignant tumors. At present, cancer is still a disease that poses a huge threat to human health. Although some cancer patients have been cured, cancer is still the first cause of cancer among residents in our country. cause of death. At present, the main treatment methods for cancer include surgery, chemotherapy, radiotherapy, targeted therapy, and traditional Chinese medicine therapy. However, the therapeutic effect of the above methods is limited, and the side effects are relatively large. In recent years, the application of cancer immunotherapy has become more and more popular, but the effect still needs to be improved. [0003] Immune checkpoint blockade (ICB) therapy can reactivate the immune system to exer...

Claims

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Application Information

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IPC IPC(8): A61K47/46A61K39/00A61K9/127A61K45/06A61P35/00A61P35/04C12N5/078
Inventor 汪超
Owner SUZHOU UNIV
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