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Gingseng extract product FQR1 and application thereof in tumour immunotherapy

A technology of ginseng extraction and FQR1-DC-CIK is applied in the field of anti-tumor activity to achieve the effects of prolonging tumor-bearing survival, promoting T cell differentiation and two-way regulation of immunity, and increasing phagocytosis and antigen presentation capabilities

Inactive Publication Date: 2016-01-13
东营凤起生物科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] The ginseng extract FQR1 of the present invention and its application in tumor cell immunotherapy have not yet seen any relevant reports at home and abroad

Method used

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  • Gingseng extract product FQR1 and application thereof in tumour immunotherapy
  • Gingseng extract product FQR1 and application thereof in tumour immunotherapy
  • Gingseng extract product FQR1 and application thereof in tumour immunotherapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] "Example 1" Preparation of Ginseng Extract FQR1

[0036] Take 500g of raw sun-dried ginseng, pulverize it with a high-speed tissue grinder, add 10 times the amount of 10mM Tris-HCl (0.15MNaCl) buffer at pH 7.4, stir well, and extract with stirring at 4°C for 12h. Centrifuge at 6000rpm for 30min, and collect the supernatant. Add 10-fold amount of 10 mM Tris-HCl (0.15M NaCl) at pH 7.4 to the precipitate for secondary leaching, collect the supernatant after centrifugation, and combine the two supernatants. Solid ammonium sulfate was added to the supernatant to a final concentration of 80%, stirred evenly and left to stand overnight at 4°C for centrifugation, the precipitate was washed 3 times with distilled water and then freeze-dried. The lyophilized samples were placed in dialysis bags, dialyzed overnight at 4°C and then lyophilized. The freeze-dried product is ginseng extract FQR1.

Embodiment 2

[0037] "Example 2" Isolation, induction and cultivation of immune cell FQR1-DC-CIK

[0038] Patient preparation: According to the indications and contraindications of immune cell FQR1-DC-CIK treatment, select tumor patients who are suitable for this treatment, inform all relevant information and precautions involved in this treatment, and obtain the understanding and cooperation of patients and their families;

[0039] Sign the informed consent form; check blood routine;

[0040] Peripheral blood mobilization: subcutaneous injection of GM-CSF150ug 24 hours before blood collection;

[0041]Peripheral blood collection: patients should have a light diet before blood collection, aseptically extract 50ml of peripheral blood from tumor patients, anticoagulate with sodium heparin and mix well to avoid coagulation;

[0042] Tumor antigen acquisition: Centrifuge at room temperature at 2000rpm for 10 minutes, collect an appropriate amount of upper plasma, inactivate and add induction m...

Embodiment 3

[0047] 《Example 3》FQR-1 induces the proliferation of DC-CIK cells

[0048] 1. Routinely separate peripheral blood mononuclear cells, and use RPMI-1640 (Shanghai Kemin Biotechnology Co., Ltd.) to adjust the cell concentration to 1×10 5 / ml, respectively inoculated in 96-well cell culture plates, 100 μl per well, 8 wells per plate, a total of three plates;

[0049] 2. Use 150 μl / well of 2× concentrated FQR1-DC-CIK cells and 2× concentrated conventionally induced DC-CIK culture solution, respectively. The first four wells of each plate are marked as the experimental group, and the last four wells are marked as the control group;

[0050] 3. Set at 37°C, 5% CO 2 Cultivate in a carbon dioxide incubator with (V / V) saturated humidity;

[0051] 4. 3 days, 5 days, and 7 days in different time periods, respectively, to detect its OD570; the results are as follows: figure 1 shown. It shows that FQR1 can effectively improve the proliferation of DC-CIK cells.

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Abstract

The invention provides a gingseng extract product FQR1 as well as a preparation and an application thereof, and provides a FQR1 induced culture method for DC-CIK cell. The method comprises the following steps: a collection of peripheral blood, an acquisition of tumor antigens, a separation of mononuclear cells, a washing, an induction of DC-CIK cells, a culture, etc. The obtained gingseng extract product FQR1 can effectively increase propagation of DC-CIK cells, improve tumor killing activity of DC-CIK cells, and prolong the survival time with tumor of tumor-bearing mice with the DC-CIK cell therapy.

Description

Technical field: [0001] The invention belongs to the technical field of immunotherapy, and in particular relates to using ginseng extract FQR-1 to induce the proliferation and differentiation of DC-CIK cells so as to improve its tumor-killing activity. Background technique: [0002] The biotherapy of tumor cells is a new tumor treatment mode with significant curative effect besides chemotherapy, radiotherapy and surgery, and it is a new treatment method for autologous immune cells to fight cancer. It has been listed by the National Health and Family Planning Commission as three types of technologies that allow clinical applications. Among them, the treatment technology of cytokine-induced killer cells (CIK / DC-CIK cells) in the peripheral blood of tumor patients has been widely used in clinic. [0003] Biological therapy includes cytokine therapy, immune cell therapy, gene therapy, molecular targeted therapy and antibody targeted therapy, etc. Its core technology is in vitr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/0783A61K36/258A61P35/00
Inventor 杨兆勇李霞张志斐汪康游白利平张紫浓李亚东金媛媛
Owner 东营凤起生物科技发展有限公司
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