Application of hemsley rockvine root extract in preparing tumor immunity therapeutic drug and human gamma delta T cell proliferation agent

An immunotherapy drug and cell proliferation technology, which is applied in the field of medicine and biology, can solve problems such as dosage inconsistencies, and achieve the effect of improving killing activity

Inactive Publication Date: 2017-02-22
浙江博纳生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This result is obviously inconsistent with the actual clinical dosage

Method used

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  • Application of hemsley rockvine root extract in preparing tumor immunity therapeutic drug and human gamma delta T cell proliferation agent
  • Application of hemsley rockvine root extract in preparing tumor immunity therapeutic drug and human gamma delta T cell proliferation agent
  • Application of hemsley rockvine root extract in preparing tumor immunity therapeutic drug and human gamma delta T cell proliferation agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Experimental situation of extraction and separation of Clover medicinal material

[0036] 1. Materials and methods

[0037] 1.1. Materials

[0038] The dry tuber root of Clover was purchased from Ningbo Yinzhou Pharmaceutical Co., Ltd.; 80% macroporous adsorption resin tree chromatograph; Yanaco micro melting point measuring instrument (the thermometer was not calibrated); Varian MAT-212 mass spectrometer; Bruker-speckospinAC-600P type nuclear magnetic resonance apparatus. Thin-layer chromatography silica gel plate and column chromatography silica gel (100-200 mesh, 200-300 mesh) are produced by Shandong Yantai Jiangyou Silica Gel Development Company; Sephadex LH-20 (20-80 μm) is produced by Pharmacia; ODS C18 reversed-phase silica gel It is produced by Merck Company; ethanol is used for extraction; petroleum ether, ethyl acetate, and n-butanol are pharmaceutical grades, water is distilled water, and the rest of the reagents are of analytical grade.

[003...

Embodiment 2

[0056] Example 2: Effect of Clover Extract on the Proliferation of γδT Cells

[0057] 1. Materials and methods

[0058] 1.1. Experimental materials

[0059] Serum-free medium (produced by Zhejiang Bona Biotechnology Co., Ltd.); tetramethylazolyl tetrazolium (MTT) and dimethyl sulfoxide (dimethyl sμlfoxide DMSO) (Sigma); lymphocyte separation solution (China Institute of Hematology, Academy of Sciences); IL-2, IL-15 and IL-21 (Xiamen Tebao Biological Company); INF-γ was purchased from Abender Company; trypsin, RPM11640, calf serum (all purchased from GIBCO Company); Human AB serum (Xuzhou Blood Station); FITC and PE-labeled CD3, FITC-labeled γδTCR, PE-labeled perforin (pore-forming protein, PFP), granzyme B (Granzyme B, GrB), APC-labeled CD107a ( Lianke Biological Co., Ltd.); Lactate dehydrogenase (Lactate dehydrogenase, LDH) kit (Japan Shino Clinical Diagnostics Co., Ltd.); Encore automatic biochemical analyzer (Beijing Xiyake Technology Co., Ltd.; fluorescence inverted micr...

Embodiment 3

[0072] Embodiment 3: Inhibition test of three leaf green extracts on three kinds of tumor cell lines

[0073] 1. Materials and methods

[0074] 1.1. Experimental materials

[0075] Serum-free medium (produced by Zhejiang Bona Biotechnology Co., Ltd.); tetramethylazolyl tetrazolium (MTT) and dimethyl sulfoxide (dimethyl sμlfoxide DMSO) (Sigma); lymphocyte separation solution (China Institute of Hematology, Academy of Sciences); Lactate dehydrogenase (Lactate dehydrogenase, LDH) kit (Japan Shino Clinical Diagnostics Co., Ltd.); Encore automatic biochemical analyzer (Beijing Xiyake Technology Co., Ltd.; fluorescence inverted microscope TE2000-S (Nikon Company, Japan); flow cytometry (BD Company, USA); flow cytometry analysis software Cell Quest, the number of cells analyzed for each sample ≥ l×104.

[0076] 1.2. Effect of Clover extract on the growth of three tumor cell lines

[0077] Breast cancer MCF-7, gastric cancer SGC-7901 and liver cancer HepG2 cells in the logarithmic ...

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Abstract

The invention relates to application of a hemsley rockvine root extract in preparing a tumor immunity therapeutic drug and a human gamma delta T cell proliferation agent, belonging to the technical field of medicine and biology. The hemsley rockvine root extract can notably accelerate proliferation of human gamma delta T cells, improves the expressions of porforin, CD107a and granzyme B of the gamma delta T cells, and has certain dose dependence; and the gamma delta T cells treated by the hemsley rockvine root extract can apparently improve the tumor-cytotoxicity activity of the gamma delta T cells and have certain dose dependence. A high-concentration hemsley rockvine root extract can inhibit the growth of tumor cells. The invention initially discovers that a low-concentration hemsley rockvine root extract can notably promote proliferation of the human gamma delta T cells; the low-concentration hemsley rockvine root extract can promote expressions of the porforin, granzyme B and CD107a of the gamma delta T cells; the cytotoxicity activity of the gamma delta T cells is apparently improved; and different extracted components of the hemsley rockvine root are apparently different in proliferation of the gamma delta T cells.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and in particular relates to the application of a trefoil extract in the preparation of tumor immunotherapy drugs and human γδT cell proliferation agents. Background technique [0002] In recent years, tumor biotherapy has become a research hotspot in the field of tumor treatment. γδT cells are an independent lymphocyte population, mainly distributed in human mucosa and epithelial tissues, have strong anti-tumor effects, and are mainly recognized by the major histocompatibility complex (MHC) in a non-restricted manner Tumors kill tumor cells through pathways such as granzyme B (granzyme B) and perforin (perforin), and interact with other innate immune cells and adaptive immune cells to help exert anti-tumor effects. Human γδT cells have become a new candidate effector cell for adoptive immunotherapy in cancer patients. It can be seen that it is very important and necessary to effec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/87A61P35/00C12N5/0783
Inventor 刘军权陈壮洪倪建芬何秋富
Owner 浙江博纳生物科技有限公司
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