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IL-2-induced T cell kinase stimulant polypeptide and use thereof

An agonist, IL-2 technology, applied in the field of peptides for the treatment of malignant tumors, can solve the problem of no lymphocyte protein tyrosine kinase agonist peptides and other problems

Inactive Publication Date: 2017-03-08
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Currently, there is no well-developed lymphocyte protein tyrosine kinase agonist polypeptide available for the treatment of malignant tumors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] IC50 of IL-2-induced T cell kinase agonist polypeptide on the growth and survival of human colon cancer cells cultured in vitro.

[0015] Using the MTT colorimetric method. The human colon cancer cell of logarithmic growth was added in a 96-well culture plate at 1.0 × 105, and cultivated for 24 hours. The experimental drug IL-2-induced T cell kinase agonist polypeptide ( Shanghai Sangong Synthetic) and the positive control drug paclitaxel; the same volume of solvent was added to the blank group. Five duplicate wells were set up in each well, and after culturing for 48 hours, MTT was added to each well. After 4 hours of action, DMSO was added, incubated for 30 minutes, and the absorbance A value was measured at 620nm in a microplate reader. According to the formula, the growth inhibition rate of human colon cancer cells = (1 - Absorbance value of the experimental group / absorbance value of the control group) × 100%. The calculated IC50 of the test drug was 5.43 μM.

Embodiment 2

[0017] IC50 of IL-2-induced T cell kinase agonist polypeptide on the growth and survival of human pancreatic cancer cells cultured in vitro.

[0018] Using the MTT colorimetric method. The logarithmic growth of human pancreatic cancer cells was added to a 96-well culture plate at a rate of 1.0×105, and cultured for 24 hours. Experimental wells and positive drug control wells were respectively added different concentrations of experimental drug IL-2-induced T cell kinase agonist polypeptide ( Shanghai Sangong Synthetic) and the positive control drug paclitaxel; the same volume of solvent was added to the blank group. Set up five duplicate wells in each well. After 48 hours of culture, MTT was added to each well. After 4 hours of action, DMSO was added, incubated for 30 minutes, and the absorbance A value was measured at 620nm on a microplate reader. According to the formula, the growth inhibition rate of human pancreatic cancer cells = (1 - Absorbance value of the experimental...

Embodiment 3

[0020] Effect of IL-2-induced T-cell kinase agonist polypeptide on human vascular endothelial cells.

[0021] Using scratch test. First use a marker pen on the back of the 24-well plate, and use a ruler to draw horizontal lines evenly, about every 0.5~1cm, across the holes. Three lines were passed through each hole; the logarithmic growth of vascular endothelial cells was added to a 24-well culture plate at 1.0×105, and cultured for 24 hours. On the second day, compare the ruler with a 10 μl pipette tip, scratch perpendicular to the horizontal line on the back, and use the intersection of the scratch and the horizontal line on the back as a fixed observation point; add different concentrations of experimental wells and positive drug control wells respectively. Drug IL-2-induced T cell kinase agonist polypeptide and positive control drug vincristine; the same volume of solvent was added to the blank group, and five replicate holes were set in each well; placed in 37 ° C, 5% CO...

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Abstract

The invention relates to the field of drugs and concretely relates to a polypeptide having the activity of stimulating IL-2-induced T cell kinase and a function of treating cancers. The polypeptide has a novel sequence, can inhibit colon cancer cells and pancreatic cancer cell proliferation activity, and can inhibit the migration of vascular endothelial cells. An in-vivo test proves that the polypeptide improves a survival rate of tumor-bearing mice and has a potential novel drug development value.

Description

technical field [0001] The invention relates to an IL-2-induced T cell kinase agonist polypeptide and its application, in particular to a polypeptide capable of promoting IL-2-induced T cell kinase activity and treating malignant tumors. Background technique [0002] IL-2-induced T cell kinase (Itk) is a member of the Tec kinase family of non-receptor tyrosine kinases, which is mainly expressed in human T cells, in the T cell receptor (TCR) and chemokine receptors important role in signal transduction pathways. Itk regulates molecular mechanisms such as phospholipase C-(PLC-7) activation, calcium mobilization, transcription factor expression and troponin recombination. Itk affects the development, differentiation and function of T cells, especially Th2 cells. Studies have found that Itk-deficient T cells exhibit a variety of developmental and functional defects, including impaired positive selection of thymocytes, an imbalance in the ratio of CD4 T / CD8 T cells, a decrease i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00A61K38/16A61P35/00
CPCA61K38/00C07K14/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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