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Pharmaceutical composition and kit for treating cancers

A technology of composition and kit, applied in the direction of drug combination, drug delivery, and medical raw materials derived from mammals, etc., can solve the problem of lack of effective means for advanced refractory solid tumors or hematological malignancies, and achieve defense against autoimmunity Escape mechanism, good application prospects, and the effect of improving clinical efficiency

Inactive Publication Date: 2017-07-18
SHUNHAO CELL BIOTECHNOLOGY (TIANJIN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is currently no effective means for the treatment of advanced refractory solid tumors or hematological malignancies in clinical practice.

Method used

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  • Pharmaceutical composition and kit for treating cancers
  • Pharmaceutical composition and kit for treating cancers
  • Pharmaceutical composition and kit for treating cancers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0045] Example 1 Acquisition of DC-CIK cells

[0046] 1. Separation of mononuclear cells from peripheral blood samples: take fresh human peripheral blood samples, add blood anticoagulants to them to prepare anticoagulant blood samples; add 1 / 3 to 2 / 3 volume of normal saline to the anticoagulant blood samples , mix thoroughly, separate the mononuclear cells in the resulting mixture with 1 to 1.5 times the volume of lymphocyte separation medium (Tianjin Haoyang, product number LTS1077006), and use lymphocyte culture medium GT-T551 (Takara, product number: DL-104 ) adjust the monocyte density to 1×10 6 ~5×10 6 cells / mL to obtain a suspension of peripheral blood mononuclear cells; wash the suspension of peripheral blood mononuclear cells with normal saline, centrifuge at room temperature for 20-30 min at a speed of 1500-2000 r / min, and obtain buffy coat cells.

[0047] 2. Separation of DC and CIK cells: Inoculate monocytes to 75cm 2 2 hours after the flask was cultured, the sus...

Embodiment 2

[0049] Example 2 Anti-PD-1 antibody activation of CIK cells in vitro experiment CIK cells stimulated by CD3 monoclonal antibody for 5 days were taken (CIK cells were cultured alone, and the immune checkpoint of CIK cells in an activated state was blocked by PD-1 antibody at this time). Broken) into 96-well flat-bottom plate, after incubation overnight, add 10μg / ml anti-PD-1 antibody and 100ng / ml tetanus toxin (TT), the control group did not add PD-1 antibody, and collected after 3 days of culture The supernatant was used to detect the secretion level of TNF-α in the PD-1 antibody group and the control group with the TNF-α ELISA detection kit from Thermofisher. Results (see figure 2 ) shows that the anti-PD-1 antibody can stimulate the function of CIK cells, and the increase rates of TNF-α secretion in the control group and the anti-PD-1 antibody group are 107.3% and 275.4%, respectively, with significant difference, p<0.01.

Embodiment 3

[0050] MTT assay of the cytotoxic effect of embodiment 3DC-CIK cells on tumor cells

[0051] The DC-CIK co-culture activated by PD-1 antibody cultured for 12 days was used as the effector cells, and the A549 lung cancer cell line was used as the target cells. Take the A549 cells in the logarithmic growth phase, and adjust the cell concentration of DC-CIK to 2.5×10 5 / mL, 5×10 5 / mL, 1×10 6 / mL, 2×10 6 / mL, the number ratio of effector cells to target cells was 2.5:1, 5:1, 10:1, 20:1. The experiment was divided into 3 groups, with 3 replicate wells in each group. The experimental group added 100 μL / well of effector cells and target cells, the target cell group added 100 μL / well of target cells and culture medium, and the effector cell group added effector cells and culture medium 100 μL / well. 100 μL / well, set at 37°C, 5% CO 2 Incubate in an incubator for 24 hours, add MTT reagent, 20 μL / well, incubate at 37°C for 4 hours, then add washing reagent 100 μL / well, incubate at 3...

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Abstract

The invention provides a pharmaceutical composition and a kit for treating cancers. The pharmaceutical composition comprises an antibody resisting a programmed cell death receptor 1 and co-cultured dendritic cells and cytokine-induced killer cells; the kit comprises the antibody resisting a programmed cell death receptor 1 and an inducer of the co-cultured dendritic cells and cytokine-induced killer cells, which are separately arranged. The pharmaceutical composition or the kit can be effectively used for treating cancers, and particularly are applicable to treatment of advanced refractory solid tumors or hematological malignancy.

Description

technical field [0001] The invention belongs to the field of cancer immunotherapy, and relates to a method for treating cancer, such as advanced refractory solid tumors or hematological malignancies, comprising anti-programmed cell death receptor 1 antibody, co-cultured dendritic cells and cytokines Pharmaceutical compositions and corresponding kits for induced killer cells, and their uses. Background technique [0002] At present, the number of cancer deaths worldwide is increasing every year, and conventional surgery, radiotherapy, and chemotherapy are not effective in treating malignant tumors. Therefore, in recent years, emerging cancer immunotherapy has attracted more and more attention and attention. Immunotherapy is becoming an important field of clinical cancer treatment, in which inhibition of immune checkpoint pathways is considered to be one of the most promising therapeutic modalities. [0003] The induction of apoptosis and anergy of tumor antigen-specific T ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P35/00A61K35/15A61K35/17
CPCA61K9/0019A61K35/15A61K35/17A61K39/39558A61K2300/00
Inventor 张云朱泽李镜李向臣李相国费晨钟殿胜王继明陈镭王立祥王华庆李忠廉刘卫平
Owner SHUNHAO CELL BIOTECHNOLOGY (TIANJIN) CO LTD
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