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Composition containing NK (natural killer) cells and antibody taking VEGF (vascular endothelial growth factor) as target and application to liver cancer

A technology of NK cells and compositions, applied to liver cancer cells MHCC97, in the field of monoclonal antibodies and/or bispecific antibodies, can solve the problems of easy recurrence and low remission rate of patients, achieve effective therapeutic effects and reduce treatment costs , The effect of simplifying the clinical treatment process

Inactive Publication Date: 2019-09-27
安徽瑞达健康产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] For various cancer diseases, using existing chemotherapy, surgical treatment, antibody therapy, etc., some patients can be relieved or achieve the purpose of disease treatment, but for a single treatment, it is still inevitable that some patients have a low remission rate or are prone to relapse And other phenomena, on this basis, medicine often adopts a combination of multiple therapies

Method used

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  • Composition containing NK (natural killer) cells and antibody taking VEGF (vascular endothelial growth factor) as target and application to liver cancer
  • Composition containing NK (natural killer) cells and antibody taking VEGF (vascular endothelial growth factor) as target and application to liver cancer
  • Composition containing NK (natural killer) cells and antibody taking VEGF (vascular endothelial growth factor) as target and application to liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The acquisition of embodiment 1 NK cells

[0042] NK cell acquisition Mononuclear cells (PBMCs) are isolated from peripheral blood and NK cells are expanded:

[0043] (1) Turn on the biological safety cabinet 30 minutes before use;

[0044] (2) Take out D-PBS from the refrigerator before use, and let it stand at room temperature for 30 minutes;

[0045] (3) Transfer 30ml of peripheral blood samples (heparin anticoagulant) to two sterile 50ml centrifuge tubes, 15ml in each tube, then add 22.5ml of sterile D-PBS to each tube, invert the centrifuge tube repeatedly, and mix thoroughly;

[0046] (4) Take another two 50ml sterile centrifuge tubes, add 15ml Ficoll-Paque Plus solution respectively, and then slowly add 24ml blood diluted in step 3 (drawn from the two sterile tubes in step 3) to form Layering, 20°C, 400×g, centrifuge for 30 minutes;

[0047] (5) Put the two 50ml centrifuge tubes in step 4 into a biological safety cabinet, then use a 10ml pipette to suck off 15...

Embodiment 2

[0060] The killing activity of embodiment 2 composition to MHCC97 cell

[0061] Preparation of cell composition: the NK cells prepared in Example 1 were prepared into 1*10 10 cells / ml of cell suspension, and then the monoclonal antibody and / or bispecific antibody targeting VEGF, preferably bevacizumab solution, was slowly added to the cell suspension, and the mixture was kept in a sterile sealed environment at 37 Incubate in a water bath at ℃ for 20 minutes, and at the same time, the cell mixture is uniformly mixed according to conventional methods, and aseptic operation must be ensured.

[0062] The final concentration or content of the bevacizumab solution mixed in the cell suspension is respectively 500 μg / ml.

[0063] Experimental group: blank control group: normal saline;

[0064] Composition group: NK cells and bevacizumab (500 μg / ml bevacizumab);

[0065] Monoclonal antibody group: bevacizumab (500 μg / ml bevacizumab);

[0066] Cell group: NK cells (1*10 10 cells / ml...

Embodiment 3

[0069] Example 3 composition inhibits the migration of MHCC97 cells

[0070] Experimental materials: (1) Transwell chamber: 24-well, 8.0-μm pore membranes (Corning);

[0071] (2) Reagents related to cell culture: serum-free medium, 10% serum medium, PBS, 0.02% EDTA;

[0072] (3) Fixative: methanol;

[0073] (4) staining solution: Giemsa dyeing solution;

[0074] (5) Mounting agent: neutral gum;

[0075] (6) Others: tweezers, cotton swabs, glass slides, coverslips;

[0076] Experimental group: the preparation method is the same as that of the cell composition in Example 2

[0077] Blank control group: normal saline;

[0078] Composition group: NK cells and bevacizumab (500 μg / ml bevacizumab);

[0079] Monoclonal antibody group: bevacizumab (500 μg / ml bevacizumab);

[0080] Cell group: NK cells (1*10 10 cells / ml of cell suspension)

[0081] experiment procedure:

[0082] (1) All cell culture reagents and Transwell chamber were incubated at 37°C;

[0083] (2) Take MHCC...

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Abstract

The invention discloses composition containing NK (natural killer) cells and an antibody taking a VEGF (vascular endothelial growth factor) as a target and an application of the composition to liver cancer. The composition specifically comprises the NK cells and a monoclonal antibody and / or bispecific antibody and is applied to liver cancer cell MHCC97 recurrent metastatic tumor. The composition containing the NK cells and the monoclonal antibody and / or bispecific antibody taking the VEGF as a target has a high killing function on MHCC97, can inhibit MHCC97 metastasis and has very good clinical application value in application of moderate and advanced anti-tumor treatment or recurrent metastatic liver cancer tumor.

Description

Technical field: [0001] The present invention relates to a composition, which includes immunotherapy cells and antibodies, specifically NK cells and VEGF-targeted monoclonal and / or bispecific antibodies, and the composition is applied to liver cancer cells MHCC97. Background technique: [0002] Cellular immunotherapy is one of the most promising tumor treatment methods at present. It can kill tumor cells through in vitro expansion or transformation and then infuse into the patient's body, or activate the immune system of the body to enhance the autoimmune function of tumor patients to resist tumors or tumors. other illnesses. At present, NK cell immunotherapy has received more and more attention. NK cells account for 5-15% of human peripheral blood lymphocytes, and their phenotype is generally defined as CD3-CD56+. NK cells can be further subdivided into two main subgroups: CD56highCD16- cells with immune regulation function and cells with Toxic active CD56dimCD16+ cells. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/17A61K39/395A61P35/00
CPCA61K35/17A61K39/395A61P35/00A61K2300/00
Inventor 邢永梅邓蒙蒙程箫刘丹吴疆王保如
Owner 安徽瑞达健康产业有限公司
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