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Immunotherapy combination medicine for treating solid tumors

An immunotherapy, solid technology, applied in the field of biomedicine

Active Publication Date: 2022-03-18
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing immune checkpoint inhibitor PD-1 / PD-L1 antibodies have little effect on Pan02-type immunotolerant pancreatic cancer solid tumors

Method used

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  • Immunotherapy combination medicine for treating solid tumors
  • Immunotherapy combination medicine for treating solid tumors
  • Immunotherapy combination medicine for treating solid tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The embodiment of the present application provides the culture and passage of HFF (human foreskin fibroblast) and Pan02 pancreatic cancer tumor cells, specifically including:

[0049] 1. Resuscitate HFF and Pan02 cells, using complete medium (10% DMEM + 10% FBS + 1% penicillin / streptomycin solution) at 37°C, 5% CO 2 Cultivated in an incubator;

[0050] 2. When the confluence of HFF or Pan02 reaches 90%, add an appropriate amount of 0.25% trypsin-EDTA to cover the cell layer on the bottom of the culture flask, digest at 37°C for 4 minutes;

[0051] 3. After the cells are suspended and detached, add complete medium to stop trypsinization, then transfer the two types of cells to a new 15ml centrifuge tube, centrifuge at 1000rpm for 5min, and discard the supernatant;

[0052] 4. Then add complete medium to resuspend the cell pellet, dilute the cell suspension according to the ratio, and transfer it to cell culture dishes of different specifications such as cell culture fla...

Embodiment 2

[0054] The embodiment of the present application provides the subculture and preparation of non-replicating uracil auxotroph Toxoplasma gondii (NRTUA strain), specifically including:

[0055] 1. Resuscitate the non-replicating uracil auxotrophic Toxoplasma gondii (NRTUA avirulent strain) for use; when the confluence of HFF cells reaches more than 90%, replace the culture medium of HFF cells with a maintenance medium containing 250 μM uracil (mass fraction is 3%DMEM+mass fraction is 10%FBS+mass fraction is 1% penicillin / streptomycin solution), then 50×10 4 NRUTA strains were added to the HFF cell culture flask;

[0056] 2. Place the medium containing HFF cells inoculated with NRUTA at 37°C, 5% CO 2 Cultivated in an incubator, NRTUA is completely broken and passaged every 4 to 5 days;

[0057] 3. After breaking the NRUTA, use a 3 μm diameter filter to filter out cell debris, collect the filtered NRTUA tachyzoites, and resuspend them in PBS to adjust the concentration of NRTUA ...

Embodiment 3

[0059] The embodiment of the present application provides the subcutaneous tumor formation experiment of Pan02 tumor cell syngeneic mice, animal grouping and administration strategies of different drugs, including:

[0060] 1. Subcutaneous tumor formation experiment of Pan02 tumor cell syngeneic mice:

[0061] 1) Purchasing animals: purchase a sufficient number of 4-6 week-old C57BL6 / J female mice from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.;

[0062] 2) Animal breeding: SPF animal room A, North Campus of Sun Yat-sen University;

[0063] 3) Collect the Pan02 cells in the logarithmic growth phase, wash twice with PBS, digest with 0.25% trypsin-EDTA, resuspend the cells in PBS and count, adjust the viable cell density to 1×10 with PBS 7 cells / ml, use a 1ml syringe to inoculate 0.2ml of Pan02 cell suspension subcutaneously in the right flank of the abdomen of C57BL6 / J mice;

[0064] 4) Measure the volume of the mouse tumor with a vernier caliper every thr...

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Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to an immunotherapy combined medicine for treating solid tumors. The invention provides an immunotherapy combined medicine and immunotherapy combined therapy for treating solid tumors. The immunotherapy combined medicine is composed of a PD-1 / PD-L1 inhibitor and a toxoplasma vaccine. The arch body is a non-toxic arch body or / and an attenuated arch body; the solid tumor is an immune tolerance solid tumor. The invention provides an immunotherapy combined drug and an immunotherapy combined therapy for treating solid tumors, and finds that the toxic toxoplasma or / and the attenuated toxoplasma are combined with a PD-1 / PD-L1 inhibitor for use, so that the solid tumors with immune tolerance can be effectively treated, and the use curative effect and the application range of the PD-1 / PD-L1 inhibitor are expanded.

Description

technical field [0001] The application belongs to the technical field of biomedicine, and in particular relates to an immunotherapy combination drug and an immune combination therapy for treating solid tumors. Background technique [0002] Immunotherapy is an emerging and effective treatment strategy for malignant tumors that are inoperable or relapsed after metastasis, but for solid tumors such as pancreatic cancer that are resistant to the immune microenvironment, PD-1 antibodies (such as αPD-1 ) Drugs usually have no obvious clinical effect or are basically ineffective. [0003] Pancreatic ductal adenocarcinoma (Pancreatic ductal adenocarcinoma, PDAC) accounts for more than 90% of pancreatic cancer (Pancreatic cancer PC), and its prognosis is extremely poor, with a 5-year overall survival rate of less than 10%. ranked second. PDAC remains a challenging disease due to its lack of early symptoms and its susceptibility to local invasion and distant metastasis, as well as r...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K39/002A61K39/395A61P35/00
CPCA61K45/06A61K39/002A61K39/3955A61P35/00A61K2300/00Y02A50/30
Inventor 周兴旺郝桃方
Owner SUN YAT SEN UNIV
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