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Application of Liraglutide in preparing tumor immnuo-therapy drugs

A technology of liraglutide and anti-tumor immunity, applied in the application field of liraglutide in the preparation of tumor immunotherapy drugs, can solve the problems of difficult to control tumor recurrence, metastasis, and inability to achieve long-term therapeutic effects.

Pending Publication Date: 2020-10-20
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional surgery, radiotherapy, chemotherapy and other programs can inhibit the progression of the tumor to a certain extent, but it is difficult to control the recurrence and metastasis of the tumor
Emerging immunotherapy in the 21st century has brought a new dawn to patients, but there are still some obstacles that prevent it from achieving long-term therapeutic effects or being more widely used in cancer treatment
So far there is no report of liraglutide used in anti-tumor immunotherapy, we found by chance that liraglutide can inhibit the activation of inflammation-related NF-κB and STAT3 pathways, and the activation of NF-κB and STAT3 pathways Activation has a strong correlation with tumor development, so we try to apply it to anti-tumor immune activation and anti-tumor immunotherapy

Method used

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  • Application of Liraglutide in preparing tumor immnuo-therapy drugs
  • Application of Liraglutide in preparing tumor immnuo-therapy drugs
  • Application of Liraglutide in preparing tumor immnuo-therapy drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Liraglutide promotes anti-tumor immunity by inhibiting inflammation in vitro

[0027] 1. Experimental materials and methods

[0028] 1.1 Experimental reagents

[0029]Human recombinant TNF-α (10620-HNAE), mouse recombinant TNF-α (50349-MNAE), human recombinant IL-6 (10395-HNAE), mouse recombinant IL-6 (50136-MNAE) were purchased from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd.

[0030] 1.2 Cell lines

[0031] Human liver cancer cells HCC-LM3 and mouse liver cancer cells Hepa1-6 were purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences, and LM3-luciferase was donated by the Institute of Hepatobiliary Surgery of Nanjing Drum Tower Hospital. Cells were cultured in DMEM containing 10% fetal bovine serum, 2 mM glutamine, 100 U / ml penicillin and 1 μg / ml streptomycin (both purchased from Invitrogen) at 37°C, 5% CO 2 , cell incubator with stable humidity.

[0032] 1.3 NKT cell culture

[0033] Isolate perip...

Embodiment 2

[0058] Example 2 Liraglutide promotes anti-tumor immunity by inhibiting tumor tissue inflammation in vivo

[0059] 1. Experimental materials and methods

[0060] 1.1 Enzyme-linked immunospot assay (ELISpot)

[0061] Note: The kit used is mouse IFN-γELISpot kit (3321-2A, Mabtech).

[0062] 1) Treat the ELISpot plate with 70% ethanol for 2 minutes, discard the ethanol, and wash 5 times with PBS.

[0063] 2) Add 100 μl of 15 μg / ml mouse IFN-γ antibody to each well, and coat overnight.

[0064] 3) Add the cell suspension to be tested and place it in a cell culture incubator for 12-48 hours.

[0065] 4) Discard the cell suspension, wash with PBS, add detection antibody, and incubate at room temperature for 2 hours.

[0066] 5) Discard the detection antibody, add streptavidin ALP, and incubate at room temperature for 1 hour.

[0067] 6) ALP was discarded, washed with PBS, and the chromogenic substrate BCIP / NBT-plus was added until spots were formed.

[0068] 7) Wash with tap w...

Embodiment 3

[0076] Example 3 NK cells mediate the effect of liraglutide in treating tumors

[0077] 1. Experimental materials and methods

[0078] 1.1 Antibody clearance experiment:

[0079] C57BL / 6 mice were divided into 4 groups, control group, treatment group, treatment group + CD8 T cell depletion group, treatment group + NK cell depletion group, 6 mice in each group. Inject 5×10 subcutaneously 6 Hepa1-6 cells, when the long diameter of the tumor reaches 0.4-0.5cm, intraperitoneally inject 500 μg / one anti-CD8α mAb (BE0017, BioXcell) to eliminate CD8 T cells, or anti-NK mAb (BE0036, BioXcell) to eliminate NK cells, and take the mouse orbit 24 hours later Venous blood was analyzed by flow cytometry to analyze the clearance rate of CD8 T cells or NK cells. After the clearance of cells was confirmed, intratumoral injection of liraglutide 300 μg / kg / mouse was started, once / day. Tumor size and body weight were measured and recorded every 3 days.

[0080] 1.2 Separation and detection of ...

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Abstract

The invention relates to the field of tumor biotherapy, particularly to application of Liraglutide in preparing immune activation drugs, anti-tumor immune activation drugs, tumor immnuo-therapy drugsand anti-tumor drugs. The invention discloses immune activation activity of the Liraglutide and anti-tumor immune activation activity thereof. The invention also discloses effects of the Liraglutide in tumor immune-therapy and tumor therapy. As shown in results according to the invention, the Liraglutide can significantly stimulate release of an interferon gamma in vitro and vivo, can significantly suppress tumor growth in a plurality of in vivo animal tumor models and significantly prolong a life cycle of tumor-bearing mice and has specific effects in wide-spectrum anti-tumor immnuo-therapy.All the results reveal application values and prospect of the Liraglutide in preparing the tumor immnuo-therapy drugs.

Description

technical field [0001] The present invention relates to the use of liraglutide for tumor immunotherapy, the use of liraglutide for inhibiting tumor-promoting inflammation, the use of liraglutide in the preparation of anti-tumor immune drugs, and the use of liraglutide in the preparation of enhanced Use in natural killer (natural killer, NK) cell function drugs. Background technique [0002] Tumor is a major disease that threatens human health. Traditional surgery, radiotherapy, chemotherapy and other programs can inhibit tumor progression to a certain extent, but it is difficult to control tumor recurrence and metastasis. The emerging immunotherapy in the 21st century has brought a new dawn to patients, but there are still some obstacles that prevent it from achieving long-term therapeutic effects or being more widely used in tumor treatment. The tumor microenvironment (tumor microenvironment, TME) is considered to be an immunosuppressive environment that cannot be ignored...

Claims

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Application Information

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IPC IPC(8): A61K38/26A61P35/00
CPCA61K38/26A61P35/00
Inventor 魏继武许纯陆贤张海林方明月梁惠董杰吴俊华
Owner NANJING UNIV
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