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Astragalus extract FQR-8 and application of astragalus extract FQR-8 in tumor cell immunotherapy

A technology of FQR-8 and Astragalus extracts, applied in animal cells, vertebrate cells, anti-tumor drugs, etc., to achieve the effects of prolonging tumor-bearing survival, increasing proliferation and reducing tumor load

Inactive Publication Date: 2015-12-16
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

Astragalus extract FQR-8 of the present invention and its application in tumor cell immunotherapy have not yet seen relevant reports at home and abroad

Method used

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  • Astragalus extract FQR-8 and application of astragalus extract FQR-8 in tumor cell immunotherapy
  • Astragalus extract FQR-8 and application of astragalus extract FQR-8 in tumor cell immunotherapy
  • Astragalus extract FQR-8 and application of astragalus extract FQR-8 in tumor cell immunotherapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] "Example 1" preparation of Astragalus extract FQR-8

[0034] Use a granulator to crush the crude drug of Astragalus membranaceus, fry it into cooked Astragalus membranaceus, add an appropriate amount of deionized water, make a slurry, add cellulase (Sinopharm Chemical Reagent Co., Ltd.) for 30 minutes, inactivate it, centrifuge at a high speed, and take the supernatant solution, and the supernatant was concentrated to make a concentrated solution, which was centrifuged again. Add ethanol to the supernatant, precipitate overnight at 4°C and centrifuge to obtain the precipitate. After the precipitate is dissolved with deionized water, it is added to the upper end of the pretreated macroporous adsorption resin XAD-16 column at a certain flow rate for adsorption. First wash slowly with deionized water to remove non-polar or water-soluble highly polar impurities such as polysaccharides or inorganic salts remaining on the surface or inside of the resin, and then use 20% meth...

Embodiment 2

[0035] "Example 2" Isolation, induction and cultivation of immune cells DC-CIK and CIK

[0036] Patient preparation: According to the indications and contraindications of immune cell DC-CIK and CIK treatment, select tumor patients who are suitable for this treatment, inform the various relevant information and precautions involved in the treatment, and obtain the understanding and cooperation of patients and their families, Sign the informed consent form and perform routine blood tests;

[0037] Peripheral blood mobilization: 24 hours before blood collection, colony-stimulating factor (GM-CSF) 150ug should be injected subcutaneously;

[0038] Peripheral blood collection: patients should have a light diet before blood collection, aseptically extract 50ml of peripheral blood from patients with tumor diseases, anticoagulate with sodium heparin and mix well to avoid coagulation;

[0039] Tumor antigen acquisition: Centrifuge at room temperature at 2000 rpm for 10 minutes, collect...

Embodiment 3

[0045] "Example 3" Induces the anti-tumor cell activity of activated FQR-DC-CIK cells

[0046] RPMI-1640 medium (Shanghai Kemin Biotechnology Co., Ltd.) adjusted 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;

[0047] Add 150ul 2×concentrated FQR-DC-CIK cells or 2×concentrated conventionally induced DC-CIK cells to each well, and at the same time, add NK cell sensitive cell line-K562 cell line (Basic Medicine, Chinese Academy of Medical Sciences) at a ratio of 1:10. Institute Cell Center). The first four wells of each plate are marked as the FQR-DC-CIK cell group, and the last four wells are marked as the conventionally induced DC-CIK cell group;

[0048] Set at 37°C, 5% CO 2 (V / V) co-cultivation in the incubator;

[0049] After culture, the OD570 tumor-killing activity was detected by MTT method;

[0050] Astragalus extract FQR-8 induced immune cell FQR-DC-CIK to k...

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Abstract

The invention provides an astragalus extract FQR-8 and a preparation method and application thereof and further provides a method for using the FQR-8 to induce and culture CIK or DC-CIK cells. The preparation method comprises the steps of peripheral blood collection, tumor antigen obtaining, mononuclear cell separation, washing, induction and culture of the DC-CIK cells and the like. The astragalus extract FQR-8 prepared by means of the preparation method can effectively promote proliferation of the CIK cells, improve the tumor killing activity of the DC-CIK cells and prolong the survival time of a tumor-bearing mouse treated by using the DC-CIK cells.

Description

Technical field: [0001] The invention belongs to the technical field of immunotherapy, and in particular relates to using astragalus extract FQR-8 to induce the proliferation and differentiation of DC-CIK cells, so as to improve the antitumor activity. Background technique: [0002] Health and life, how to improve the quality of life of malignant tumor patients has become a hot topic in this field. Malignant tumors have become the number one cause of death in our country, seriously threatening people's health. [0003] 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 is listed as the three types of technology that allow clinical application by the National Health and Family Planning Commission. Biological therapy includes cytokine therapy, immune cell therapy, gene therapy, molecular targeted therapy and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/0783A61K36/481A61P35/00
Inventor 杨兆勇李霞许乐幸汪康游张志斐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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